Effects of Vitamin E and D on the Stiffness of MCF-7

Effects of Vitamin E and D on the Stiffness of MCF-7 | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effects of Vitamin E and D on the Stiffness of MCF-7 Armin Jarahi Khameneh, Ashkan Heydarian, Nasibeh Babaei, Negin Hannani, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2313700/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Several investigations have demonstrated that vitamins can be used to treat or prevent cancer by altering actin filaments, inhibiting cell migration and cancer cell proliferation. Vitamins D and E are fat-soluble. This research aims to determine the short-term impact of vitamin D and E on the mechanical characteristics of breast cancer cells before comparing them with normal breast cells. Method Atomic force microscopy (AFM) was used to examine the deformation of MCF-10 normal breast cells, MCF-7 breast cancer cells, and MCF-7 breast cancer cells treated with 0.03 µM vitamin D and 16 µM vitamin E solution. Young's modulus was calculated employing the Hertz model to determine cell stiffness. Results The Young's modulus of vitamin D-treated cancer cells (585.8 Pa) was substantially similar to that of normal cells (455.6 Pa). Nevertheless, vitamin E treatment had no effect on Young's modulus of cancer cells, which remained remarkably similar to that of untreated cancer cells (216.6 and 203.4 Pa, respectively). Conclusion Despite vitamin E, vitamin D enhances the stiffness of tumor cells and puts their mechanical characteristics similar to normal cells by interfering with actin filaments and cell skeletons, which may inhibit tumor cell migration. According to these findings, Vitamin D appears to be an effective drug for cancer treatment. Breast Cancer Vitamin D Vitamin E Elasticity Epithelial Cell Cell Mechanics Figures Figure 1 Figure 2 Introduction Breast cancer develops when epithelial cells multiply uncontrollably and create a mass of tissue known as a tumor. Breast cancer affects a large number of people today. Breast cancer is anticipated to be the most prevalent cancer in the United States in 2022 [1]. This disease was acknowledged globally as the leading cause of mortality among women [2]. The prevention and treatment of breast cancer can benefit from using vitamins, according to investigations [3]. Both normal mammary glands and breast cancer cells have been found to express vitamin D receptors. To immunize the patient against breast cancer, vitamin D stimulates the vitamin D receptor and functions as a master transcriptional regulator of autophagy [4]. Numerous pre-clinical investigations have suggested that the 1,25D ligand plays a function in limiting normal breast gland growth and vulnerability to carcinogenesis. According to several investigations, vitamin D insufficiency is widespread in patients experiencing breast cancer, which enhances the risk of cancer development [5]. Yao et al. [6] discovered that higher levels of serum 25-hydroxyvitamin D decrease the risk of mortality and cancer among 1666 cancer patients, and this result was more significant in premenopausal women. In addition, studies evaluating the impact of vitamin D on transgenic mice models indicated that vitamin D could inhibit numerous distinctive mammary tumor models, some of which are highly comparable to human breast cancer [7–13]. Both vitamin D receptor and Cyp27b1 deletion were found to promote tumorigenesis. According to research by Ooi et al. [14] osteolytic lesions manifest themselves faster in vitamin D-deficient rats than in vitamin D-sufficient rats. When doxorubicin is combined with vitamin E, the survival of breast cancer cells is significantly shortened with minimum damage to normal cells, as demonstrated by Ahmadi et al. [15]. Despite the fact that a high dose of vitamin E exacerbates the cytotoxicity induced by doxorabicin. Sigounas et al. [16] discovered that vitamin E reduces cellular proliferation in breast and prostate tumor cells by lowering deoxyribonucleic acid (DNA) synthesis in cell lines. Additionally, substantial DNA fragmentation was seen while examining high molecular weight samples, suggesting that the vitamin E-supplemented cell lines underwent apoptosis. Nevertheless, several investigations have found that the administration of vitamin E does not affect the incidence of breast cancer [17–21]. Stiffness, nonlinearity, anisotropy, and heterogeneity are mechanical characteristics of cells. Numerous biological processes, including growth, adhesion, motility, deformation, and cell differentiation, are regulated by these characteristics [22–25]. Alterations in cellular and extracellular mechanical characteristics during cancer development alter forces on cells and influence tumor cells' morphogenetic evolution, invasion, and proliferation [26, 27]. Evaluating the mechanical characteristics of the tumor and normal cells is one technique to differentiate them. Since the mechanical characteristics and cytoskeleton structure are altered by cell carcinogenesis [28]. For instance, normal cells are less fluid [29] and more rigid than tumor cells [30–32]. In addition, the stiffness of tumor cell lines tends to decrease as the severity of cancer enhances [33]. According to Plodinec et al. [34], malignant breast tissues differ from normal tissues in that they have a wider dispersion of elastic modulus and a lower stiffness peak. Tumor cells increased the contraction of the actomyosin cortex when stretched by external stimuli compared to the related normal cells [35, 36]. Additionally, there are alterations in how tumor cells interact physically with their adjacent cells and the extracellular components surrounding them, affecting the tumors' growth rate, morphology, and invasion [37, 38]. Alterations in the mechanical characteristics of cells are accompanied by alterations in the mechanical characteristics of the extracellular environment. The majority of the extracellular milieu of the tumor is made up of fibrous tissue. As breast cancer progresses, the cross-linking between these fibers grows, making the extracellular matrix environment rigid [39]. Additionally, timely detection [40], cancer prevention [41], and measuring the efficacy of anti-cancer medications can all benefit from the distinction between normal and malignant cells [31, 42]. AFM is one of the methods that may be employed to study cell mechanical characteristics [43]. AFM is well recognized as a useful method for investigating biological materials[44–46]. It is also a versatile technique for non-destructively imaging and quantifying the elastic characteristics of living cells under physiological settings [47–51]. The contact force and distance between the tip and the sample (tip-sample) are calculated using the AFM technique when the tip moves up and down at a point. Young's modulus, which provides a quantitative value of the cellular elastic characteristics, is used to represent deformation in this method. The contact-mode point-spectroscopy curve, also known as the force curve, can be used to calculate Young's modulus. Other mechanical characteristics of materials, including adhesion, deformation, and dissipation, can be determined by employing these force curves in conjunction with Young's modulus. Nevertheless, attaining these values precisely may be challenging [52]. Ex vivo mechanical characteristics of breast cancer cells, bladder cells, and fibroblasts have been studied using this method thus far [25, 53–57]. Although numerous investigations have been undertaken to examine the influence of vitamins on breast cancer healing, only several investigations have been carried out to study the impact of vitamins E and D on the mechanical characteristics of breast cancer cells. This investigation was carried out in order to determine how vitamins D and E affect the mechanical characteristics of malignant breast tissues. Materials And Methods Two cell lines, normal breast cell MCF-10 and epithelial breast cancer cell MCF-7, were obtained in two 25 ml flasks from the Stem cell Technology Research Center. The flasks were used for defreezing. The flasks were then placed inside the incubator, and after 24 hours, the cell flasks were substituted with 5 ml of Dulbecco's Modified Eagle Medium (DMEM) high Glucose culture medium comprising 10% fetal bovine serum (FBS). An inverted microscope was used to inspect the cells' density and morphology. Once they reached the appropriate density, the cells adhered to the bottom of the cell flask were detached and seeded in the suitable 3.5 cm Petri dishes. Each cell flask received 1 ml of warmed Phosphate Buffered Saline (PBS) EDTA, enough to cover the bottom of the flask. The flasks were then filled with 1 ml of warmed trypsin enzyme (37°C) and placed in the incubator for 2 minutes. The MCF7 cell-cell line was subsequently cultured in three 3.5 cm Petri dishes, while the MCF10 cell line was cultured in another 3.5 cm petri dish, and the Petri dishes were then placed in an incubator to achieve the necessary temperature, moisture, and CO 2 . Vitamins D3 and E were purchased in two vials from the Osvah pharmaceutical Company Vitamin D3 at 0.03 U/ml, and vitamin E at 16 U/ml concentrations were calculated and prepared to achieve the necessary volume of culture media utilized in this study. The cells were treated after 24 hours of cell culture in Petri dishes. After thoroughly removing the culture medium from the Petri dishes, 3 ml of 10% FBS culture medium comprising vitamin D3 at a concentration of 0.03 U/ml was poured into one of the Petri dishes incorporating the MCF7 cell line. Another petri dish comprising the MCF7 cell line received 3 ml of 10% FBS culture medium incorporating vitamin E at a concentration of 16U/ml. The third petri dish comprising the MCF7 cell line received 3 ml of culture medium containing 10% FBS. In addition, the supernatant in the petri dish holding the MCF10 cell line was eliminated entirely, 3 ml of 10% FBS culture medium was poured, and the dish was placed in the incubator. The appropriate four Petri dishes were brought to the laboratory for the remaining steps after receiving the treatment for 24 hours. The mechanical characteristics of the cell were evaluated using a BioAFM device manufactured by JPK brand, Germany, in this investigation. A pyramidal probe containing a square base with a pyramid apex size of 7 nm and a 35-degree angle was employed in this study. The spring constant is 0.19 Nm. The apparatus was sterilized in accordance with the protocol before initiation of the UV radiation test, and the apparatus was then calibrated. The maximum contact force of 2,700 piconewtons was exerted on 25 sites of the cells in the Petri dishes during this experiment. The force-displacement curves of 25 individual cells in each group were subsequently inspected, and the force-displacement curve was drawn to record the deformation curve. All of the test steps were conducted in 2 hours and 30 minutes at 37°C, and finally, the data was extracted. The Young's modulus of cell specimens was utilized to measure cell stiffness in this research, and Young's modulus was measured using the Hertz model by analyzing the data employing the MCF10, MCF7, MCF7 + VITAMIN D3, and MCF7 + VITAMIN E cells' force-deformation diagram. \(F=\frac{E}{1-{\nu }^{2}}\frac{\text{tan}\alpha }{\sqrt{2}}{\delta }^{2}\) Eq. 1 In the probe formula, α represents the half angle to the probe's face (55 °), ϑ represents Poisson's ratio in δ-Phase, which is believed to be 0.5 for soft biological samples and denotes the deformation depth, F indicates the force on the cell, and E represents Young's modulus. Each cell's Young's modulus was determined. The data from four groups were averaged, and two groups treated with vitamin D and E were compared to MCF-10 and MCF-7 control groups. The normality test was employed to determine the non-randomness of the data collected from the Mini Tab software. The one-way ANOVA test was used to analyze the average of the four groups. Results The cells in each group's deformation were analyzed. Initially, the console's force and the AFM tip displacement were both assumed to be zero. Subsequently, for each cell in each group, the force-indentation data were analyzed, and the mean force-indentation data for all four groups were drawn (Fig. 1 ). Each group's loading and unloading processes are shown in Fig. 1 . MCF-10 cells, MCF-7 tumor cells, MCF-7 tumor cells treated with vitamin D, and MCF-7 tumor cells treated with vitamin E had maximum membrane deformations of 7.318, 0.293, 7.821 and 0.382 µm, respectively. Young's modulus and the Hertz model were determined for all four cell lines. MCF10 cells, MCF7 treated with vitamin D, MCF7 treated with vitamin E, and MCF7 had mean values and SD standard deviations of 455.6 ± 175.7, 585.8 ± 138, 216.6 ± 40.82, and 203.4 ± 43.1 Pa, respectively (Fig. 2 ). ANOVA test with a 95% confidence interval was used to compare the averages of the four groups and determine their difference. A statistically significant difference was found in the average of these groups (P < 0.05). Tukey's test for post-Hoc analysis was subsequently performed to assess the statistical significance and pairwise comparison of cell groups. Young's modulus was shown to differ across MCF10, MCF-7, MCF7 + E, and MCF7 + D3 groups. Discussion The mechanical characteristics of normal breast epithelial cells were compared with cancer cells treated with vitamin D and E in this research to evaluate the short-term (for 24 hours) impact of vitamins D and E on the mechanical properties of tumor cells. The Young's modulus acquired from the Hertz model of tumor cells treated with vitamin D was substantially similar to Young's modulus of normal cells, according to the results of this research. Nevertheless, supplementing vitamin E did not affect the characteristics of tumor cells, and Young's modulus stayed nearer to cancer cells rather than normal cells. Tumor cells become more rigid as a result of vitamin D supplementation, approaching the normal cells' stiffness. According to research, normal cells are stiffer than malignant cells [30, 32, 58]. The stiffness of the cells decreases as the malignancy progresses [19]. According to Li et al. [58, 59], decreased stiffness and a notable reduction of F-actin filaments or their bundles (stress fibers) may be the cause of the cancer cells' easier motility and invasion during metastasis. As a result, by stiffening tumor cells, vitamin D can limit metastasis and migration. The mechanical characteristics of the cells were unaffected by vitamin E treatment. There was no correlation between vitamin E and a lower risk of breast cancer in other investigations [18, 19]. Hunter et al. [20] also reported that high vitamin E supplementation did not protect women from breast cancer. Because vitamin E is lipophilic [60], various investigations have revealed that it has the potential to affect the biophysical characteristics of membrane and lipid species. Asif et al. [61] discovered that administering vitamin E as an anti-tumor medication induced substantial DNA damage, mitochondrial depolarization, apoptosis, and arrest of tumor cell proliferation. Additionally, it has been shown that treatment with TC6OH, a vitamin E derivative, induces tumor cell apoptosis by transport, breakdown, and lowering of some anti-apoptotic and growth factor-related proteins, damaging the mitochondrial membrane potential and modulating lipids [62]. Further research is proposed to evaluate the effect of the vitamins D, and E on tumor cells and that tumor cells are treated with varying doses of these vitamins to achieve more precise and reliable findings. This can respond to the question of how many doses of these vitamins can be more effective or even detrimental. It is important to note that the long-term impact of these vitamins on tumor cells is a topic that can be discussed in the future. Conclusion In summary, this research demonstrates that despite vitamin E, vitamin D has a major impact on altering the mechanical characteristics (stiffness) of tumor cells and can make them closer to the normal cells' properties, suggesting that vitamin D may be helpful in the treatment of cancer. It can also decrease the possibility of cell migration by enhancing cell stiffness. List Of Abbreviations Abbreviation Definition AFM Atomic force microscopy DNA Deoxyribonucleic acid (DMEM) Dulbecco's Modified Eagle Medium FBS Fetal Bovine Serum PBS Phosphate Buffered Saline Declarations Ethics approval and consent to participate: Not applicable. This article does not contain any studies with human participants or animals performed by any of the authors. Consent for publication: All authors consent for publication. Availability of data and material: Not applicable. Competing interests: The authors of the manuscript hereby declare that they have no conflict of interest Funding: There was no funding included. Authors' contributions: Armin Jarahi, Ashkan Heydarian and Dornaz Millani were performed the experimental testing, modeling and statistical analysis Negin Hannani and Nasibe Babaei wrote the manuscript. Hosein Sahbafar were performed cell culturing.All authors reviewed the manuscript. Acknowledgements: Not applicable. References Siegel, R. L., Miller, K. D., Fuchs, H. E., & Jemal, A. (2022). Cancer statistics, 2022. CA: a cancer journal for clinicians , 72 (1), 7–33. https://doi.org/10.3322/CAAC.21708 Sung, H., Ferlay, J., Siegel, R. L., Laversanne, M., Soerjomataram, I., Jemal, A., & Bray, F. (2021). Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA: A Cancer Journal for Clinicians , 71 (3), 209–249. https://doi.org/10.3322/CAAC.21660 Santos, J. M., Khan, Z. S., Munir, M. T., Tarafdar, K., Rahman, S. M., & Hussain, F. (2018). Vitamin D3 decreases glycolysis and invasiveness, and increases cellular stiffness in breast cancer cells. The Journal of Nutritional Biochemistry , 53 , 111–120. https://doi.org/10.1016/J.JNUTBIO.2017.10.013 Tavera-Mendoza, L. E., Westerling, T., Libby, E., Marusyk, A., Cato, L., Cassani, R., … Brown, M. (2017). Vitamin D receptor regulates autophagy in the normal mammary gland and in luminal breast cancer cells. Proceedings of the National Academy of Sciences of the United States of America , 114 (11), E2186–E2194. https://doi.org/10.1073/PNAS.1615015114 molecular, J. W.-T. J. of steroid biochemistry and, & 2018, undefined. (n.d.). Vitamin D and breast cancer: Past and present. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S0960076017301905 Yao, S., Kwan, M. L., Ergas, I. J., Roh, J. M., Cheng, T. Y. D., Hong, C. C., … Kushi, L. H. (2017). Association of Serum Level of Vitamin D at Diagnosis With Breast Cancer Survival: A Case-Cohort Analysis in the Pathways Study. JAMA Oncology , 3 (3), 351–357. https://doi.org/10.1001/JAMAONCOL.2016.4188 So, J. Y., Wahler, J. E., Yoon, T., Smolarek, A. K., Lin, Y., Shih, W. J., … Suh, N. (2013). Oral administration of a Gemini vitamin D analog, a synthetic triterpenoid and the combination prevents mammary tumorigenesis driven by ErbB2 overexpression. Cancer Prevention Research , 6 (9), 959–970. https://doi.org/10.1158/1940-6207.CAPR-13-0087/35984/AM/ORAL-ADMINISTRATION-OF-A-GEMINI-VITAMIN-D-ANALOG-A Milliken, E. L., Zhang, X., Flask, C., Duerk, J. L., MacDonald, P. N., & Keri, R. A. (2005). EB1089, a vitamin D receptor agonist, reduces proliferation and decreases tumor growth rate in a mouse model of hormone-induced mammary cancer. Cancer Letters , 229 (2), 205–215. https://doi.org/10.1016/J.CANLET.2005.06.044 Lee, H. J., So, J. Y., DeCastro, A., Smolarek, A., Paul, S., Maehr, H., … Suh, N. (2010). Gemini vitamin D analog suppresses ErbB2-positive mammary tumor growth via inhibition of ErbB2/AKT/ERK signaling. The Journal of Steroid Biochemistry and Molecular Biology , 121 (1–2), 408–412. https://doi.org/10.1016/J.JSBMB.2010.03.053 Zinser, G. M., & Welsch, J. E. (2004). Vitamin D receptor status alters mammary gland morphology and tumorigenesis in MMTV-neu mice. Carcinogenesis , 25 (12), 2361–2372. https://doi.org/10.1093/CARCIN/BGH271 Li, J., Luco, A. L., Ochietti, B., Fadhil, I., Camirand, A., Reinhardt, T. A., … Kremer, R. (2016). Tumoral Vitamin D Synthesis by CYP27B1 1-α-Hydroxylase Delays Mammary Tumor Progression in the PyMT-MMTV Mouse Model and Its Action Involves NF-κB Modulation. Endocrinology , 157 (6), 2204–2216. https://doi.org/10.1210/EN.2015-1824 Li, J., Luco, A. L., Huang, D. C., Reinhardt, T. A., Muller, W., & Kremer, R. (2015). Chemoprevention activity of 25-hydroxyvitamin d in the MMTV-PyMT mouse model of breast cancer. Cancer Prevention Research , 8 (2), 120–128. https://doi.org/10.1158/1940-6207.CAPR-14-0110/36341/AM/CHEMOPREVENTION-ACTIVITY-OF-25-HYDROXYVITAMIN-D-IN Johnson, A. L., Zinser, G. M., & Waltz, S. E. (2015). Vitamin D3-dependent VDR signaling delays ron-mediated breast tumorigenesis through suppression of β-catenin activity. Oncotarget , 6 (18), 16304. https://doi.org/10.18632/ONCOTARGET.4059 Ooi, L. L., Zhou, H., Kalak, R., Zheng, Y., Conigrave, A. D., Seibel, M. J., & Dunstan, C. R. (2010). Vitamin D deficiency promotes human breast cancer growth in a murine model of bone metastasis. Cancer Research , 70 (5), 1835–1844. https://doi.org/10.1158/0008-5472.CAN-09-3194/655774/P/VITAMIN-D-DEFICIENCY-PROMOTES-HUMAN-BREAST-CANCER Ahmadi, M., Hedayatizadeh-Omran, A., Alizadeh-Navaei, R., Saeedi, M., Zaboli, E., Amjadi, O., … Besharat, Z. (2022). Effects of Vitamin E on Doxorubicin Cytotoxicity in Human Breast Cancer Cells in Vitro. Asian Pacific Journal of Cancer Prevention , 23 (1), 201–205. https://doi.org/10.31557/APJCP.2022.23.1.201 Sigounas, G., Anagnostou, A., & Steiner, M. (2009). dl‐α‐tocopherol induces apoptosis in erythroleukemia, prostate, and breast cancer cells. http://dx.doi.org/10.1080/01635589709514549 , 28 (1), 30–35. https://doi.org/10.1080/01635589709514549 Sato, R., Helzlsouer, K., Alberg, A., … S. H.-C. E., & 2002, undefined. (2002). Prospective study of carotenoids, tocopherols, and retinoid concentrations and the risk of breast cancer. AACR , 11 , 451–457. Retrieved from https://aacrjournals.org/cebp/article-abstract/11/5/451/166599 Kushi, L., Fee, R., Sellers, T., … W. Z.-A. journal of, & 1996, undefined. (1996). Intake of vitamins A, C, and E and postmenopausal breast cancer: the Iowa Women’s Health Study. academic.oup.com , 144 (2). Retrieved from https://academic.oup.com/aje/article-abstract/144/2/165/112184 Rohan, T. E., Howe, G. R., Friedenreich, C. M., Jain, M., & Miller, A. B. (1993). Dietary fiber, vitamins A, C, and E, and risk of breast cancer: a cohort study. Cancer Causes and Control , 4 (1), 29–37. https://doi.org/10.1007/BF00051711 Hunter, D. J., Manson, J. E., Colditz, G. A., Stampfer, M. J., Rosner, B., Hennekens, C. H., … Willett, W. C. (1993). A Prospective Study of the Intake of Vitamins C, E, and A and the Risk of Breast Cancer. New England Journal of Medicine , 329 (4), 234–240. https://doi.org/10.1056/NEJM199307223290403 Zhang, S., Hunter, D., … M. F.-J. of the, & 1999, undefined. (n.d.). Dietary carotenoids and vitamins A, C, and E and risk of breast cancer. academic.oup.com . Retrieved from https://academic.oup.com/jnci/article-abstract/91/6/547/2549364 Zhu, C., Bao, G., engineering, N. W.-A. review of biomedical, & 2000, undefined. (n.d.). Cell mechanics: mechanical response, cell adhesion, and molecular deformation. researchgate.net . Retrieved from https://www.researchgate.net/profile/Cheng-Zhu-32/publication/11652210_C_ELL_M_ECHANICS_Mechanical_Response_Cell_Adhesion_and_Molecular_Deformation/links/53da1d8f0cf2631430c7f843/C-ELL-M-ECHANICS-Mechanical-Response-Cell-Adhesion-and-Molecular-Deformation.pdf?_sg%5B0%5D=started_experiment_milestone&origin=journalDetail chemistry, E. E.-A. review of biophysics and biophysical, & 1988, undefined. (1988). Cellular mechanics as an indicator of cytoskeletal structure and function. researchgate.net . https://doi.org/10.1146/annurev.bb.17.060188.002145 Pourati, J., Maniotis, A., Spiegel, D., Schaffer, J. L., Butler, J. P., Fredberg, J. J., … Wang, N. (1998). Is cytoskeletal tension a major determinant of cell deformability in adherent endothelial cells? American Journal of Physiology - Cell Physiology , 274 (5 43-5). https://doi.org/10.1152/AJPCELL.1998.274.5.C1283 Rotsch, C., Jacobson, K., & Radmacher, M. (1999). Dimensional and mechanical dynamics of active and stable edges in motile fibroblasts investigated by using atomic force microscopy. Proceedings of the National Academy of Sciences of the United States of America , 96 (3), 921–926. https://doi.org/10.1073/PNAS.96.3.921 Huang, S., cell, D. I.-C., & 2005, undefined. (n.d.). Cell tension, matrix mechanics, and cancer development. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S1535610805002679 Lopez, J. I., Mouw, J. K., & Weaver, V. M. (2008). Biomechanical regulation of cell orientation and fate. Oncogene 2008 27:55 , 27 (55), 6981–6993. https://doi.org/10.1038/onc.2008.348 Suresh, S., Spatz, J., Mills, J. P., Micoulet, A., Dao, M., Lim, C. T., … Seufferlein, T. (2005). Connections between single-cell biomechanics and human disease states: gastrointestinal cancer and malaria. Acta Biomaterialia , 1 (1), 15–30. https://doi.org/10.1016/J.ACTBIO.2004.09.001 Rother, J., Nöding, H., Mey, I., & Janshoff, A. (2014). Atomic force microscopy-based microrheology reveals significant differences in the viscoelastic response between malign and benign cell lines. Open Biology , 4 (MAY). https://doi.org/10.1098/RSOB.140046 Coughlin, M. F., Bielenberg, D. R., Lenormand, G., Marinkovic, M., Waghorne, C. G., Zetter, B. R., & Fredberg, J. J. (2012). Cytoskeletal stiffness, friction, and fluidity of cancer cell lines with different metastatic potential. Clinical & Experimental Metastasis 2012 30:3 , 30 (3), 237–250. https://doi.org/10.1007/S10585-012-9531-Z Guck, J., Schinkinger, S., Lincoln, B., Wottawah, F., journal, S. E.-B., & 2005, undefined. (n.d.). Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349505734172 Swaminathan, V., Mythreye, K., Tim O’Brien, E., Berchuck, A., Blobe, G. C., & Superfine, R. (2011). Mechanical Stiffness grades metastatic potential in patient tumor cells and in cancer cell lines. Cancer Research , 71 (15), 5075–5080. https://doi.org/10.1158/0008-5472.CAN-11-0247/649738/AM/MECHANICAL-STIFFNESS-GRADES-METASTATIC-POTENTIAL Suresh, S. (2007). Biomechanics and biophysics of cancer cells. Acta Biomaterialia , 3 (4), 413–438. https://doi.org/10.1016/J.ACTBIO.2007.04.002 Plodinec, M., Loparic, M., Monnier, C. A., Obermann, E. C., Zanetti-Dallenbach, R., Oertle, P., … Schoenenberger, C. A. (2012). The nanomechanical signature of breast cancer. Nature Nanotechnology 2012 7:11 , 7 (11), 757–765. https://doi.org/10.1038/nnano.2012.167 Jonas, O., Mierke, C. T., & Käs, J. A. (2011). Invasive cancer cell lines exhibit biomechanical properties that are distinct from their noninvasive counterparts. Soft Matter , 7 (24), 11488–11495. https://doi.org/10.1039/C1SM05532A Kraning-Rush, C. M., Califano, J. P., & Reinhart-King, C. A. (2012). Cellular Traction Stresses Increase with Increasing Metastatic Potential. PLOS ONE , 7 (2), e32572. https://doi.org/10.1371/JOURNAL.PONE.0032572 Paredes, J., Albergaria, A., Oliveira, J. T., Jeronimo, C., Milanezi, F., & Schmitt, F. C. (2005). P-Cadherin Overexpression Is an Indicator of Clinical Outcome in Invasive Breast Carcinomas and Is Associated with CDH3 Promoter Hypomethylation. Clinical Cancer Research , 11 (16), 5869–5877. https://doi.org/10.1158/1078-0432.CCR-05-0059 Ribeiro, A. S., Albergaria, A., Sousa, B., Correia, A. L., Bracke, M., Seruca, R., … Paredes, J. (2009). Extracellular cleavage and shedding of P-cadherin: a mechanism underlying the invasive behaviour of breast cancer cells. Oncogene 2010 29:3 , 29 (3), 392–402. https://doi.org/10.1038/onc.2009.338 Ansardamavandi, A., Tafazzoli-Shadpour, M., Omidvar, R., & Jahanzad, I. (2016). Quantification of effects of cancer on elastic properties of breast tissue by Atomic Force Microscopy. Journal of the Mechanical Behavior of Biomedical Materials , 60 , 234–242. https://doi.org/10.1016/J.JMBBM.2015.12.028 Lee, G., biotechnology, C. L.-T. in, & 2007, undefined. (n.d.). Biomechanics approaches to studying human diseases. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S0167779907000212 Katira, P., Bonnecaze, R. T., & Zaman, M. H. (2013). Modeling the mechanics of cancer: Effect of changes in cellular and extra-cellular mechanical properties. Frontiers in Oncology , 3 JUN . https://doi.org/10.3389/FONC.2013.00145/FULL Wottawah, F., Schinkinger, S., Lincoln, B., biomaterialia, S. E.-A., & 2005, undefined. (n.d.). Characterizing single suspended cells by optorheology. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S1742706105000589 Binnig, G., Quate, C. F., & Gerber, C. (1986). Atomic force microscope. Physical Review Letters , 56 (9), 930–933. https://doi.org/10.1103/PHYSREVLETT.56.930 Kasas, S., Thomson, N. H., Smith, B. L., Hansma, P. K., Miklossy, J., & Hansma, H. G. (1996). Biological Applications of the AFM: From Single Molecules to Organs. https://doi.org/10.1002/(SICI)1098-1098(1997)8:2 science, E. H.-P. in surface, & 1994, undefined. (n.d.). Imaging of living cells by atomic force microscopy. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/007968169490006X Hansma, H., and, J. H.-A. review of biophysics, & 1994, undefined. (n.d.). Biomolecular imaging with the atomic force microscope. jhu.pure.elsevier.com . Retrieved from https://jhu.pure.elsevier.com/en/publications/biomolecular-imaging-with-the-atomic-force-microscope-4 Schoenenberger, C., journal, J. H.-B., & 1994, undefined. (n.d.). Slow cellular dynamics in MDCK and R5 cells monitored by time-lapse atomic force microscopy. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349594805569 A-Hassan, E., Heinz, W. F., Antonik, M. D., D’Costa, N. P., Nageswaran, S., Schoenenberger, C. A., & Hoh, J. H. (1998). Relative Microelastic Mapping of Living Cells by Atomic Force Microscopy. Biophysical Journal , 74 (3), 1564–1578. https://doi.org/10.1016/S0006-3495(98)77868-3 Schaus, S., journal, E. H.-B., & 1997, undefined. (n.d.). Cell viability and probe-cell membrane interactions of XR1 glial cells imaged by atomic force microscopy. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349597781530 Haydon, P. G., Lartius, R., Parpura, V., & Marchese-Ragona, S. P. (1996). Membrane deformation of living glial cells using atomic force microscopy. Journal of Microscopy , 182 (2), 114–120. https://doi.org/10.1046/J.1365-2818.1996.141423.X Henderson, E., Haydon, P. G., & Sakaguchi, D. S. (1992). Actin filament dynamics in living glial cells imaged by atomic force microscopy. Science , 257 (5078), 1944–1946. https://doi.org/10.1126/SCIENCE.1411511 Polytech Elec Eng Comp Sci Kámán, P. J., & Kámán, J. (2015). Young’s Modulus and Energy Dissipation Determination Methods by AFM, with Particular Reference to a Chalcogenide Thin Film. Periodica Polytechnica Electrical Engineering and Computer Science , 59 (1), 18–25. https://doi.org/10.3311/PPEE.7865 Lekka, M., Laidler, P., Ignacak, J., Łabędź, M., … J. L.-… et B. A. (BBA, & 2001, undefined. (n.d.). The effect of chitosan on stiffness and glycolytic activity of human bladder cells. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S0167488901001252 Rotsch, C., journal, M. R.-B., & 2000, undefined. (n.d.). Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts: an atomic force microscopy study. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349500766148 Lekka, M., Laidler, P., Gil, D., Lekki, J., … Z. S.-E. B., & 1999, undefined. (1999). Elasticity of normal and cancerous human bladder cells studied by scanning force microscopy. Springer , 28 (4), 312–316. https://doi.org/10.1007/s002490050213 Park, S., Koch, D., Cardenas, R., Käs, J., journal, C. S.-B., & 2005, undefined. (n.d.). Cell motility and local viscoelasticity of fibroblasts. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349505730733 Cross, S. E., Jin, Y.-S., Rao, J., & Gimzewski, J. K. (2020). Nanomechanical Analysis of Cells from Cancer Patients. Nano-Enabled Medical Applications , 547–566. https://doi.org/10.1201/9780429399039-18/NANOMECHANICAL-ANALYSIS-CELLS-CANCER-PATIENTS-SARAH-CROSS-YU-SHENG-JIN-JIANYU-RAO-JAMES-GIMZEWSKI Li, Q. S., Lee, G. Y. H., Ong, C. N., & Lim, C. T. (2009). Probing the Elasticity of Breast Cancer Cells Using AFM. IFMBE Proceedings , 23 , 2122–2125. https://doi.org/10.1007/978-3-540-92841-6_530 Li, Q. S., Lee, G. Y. H., Ong, C. N., & Lim, C. T. (2008). AFM indentation study of breast cancer cells. Biochemical and Biophysical Research Communications , 374 (4), 609–613. https://doi.org/10.1016/J.BBRC.2008.07.078 Wang, X., & Quinn, P. J. (2000). The location and function of vitamin E in membranes (review). Molecular membrane biology , 17 (3), 143–156. https://doi.org/10.1080/09687680010000311 Itoo, A., Paul, M., Ghosh, B., Polymers, S. B.-C., & 2022, undefined. (n.d.). Oxaliplatin delivery via chitosan/vitamin E conjugate micelles for improved efficacy and MDR-reversal in breast cancer. Elsevier . Retrieved from https://www.sciencedirect.com/science/article/pii/S0144861722000121 Gok, S., Kuzmenko, O., Babinskyi, A., & Severcan, F. (2021). Vitamin E Derivative with Modified Side Chain Induced Apoptosis by Modulating the Cellular Lipids and Membrane Dynamics in MCF7 Cells. Cell Biochemistry and Biophysics 2021 79:2 , 79 (2), 271–287. https://doi.org/10.1007/S12013-020-00961-Y Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2313700","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":156798228,"identity":"24546a4f-9b5f-46ae-b220-784ad6600946","order_by":0,"name":"Armin Jarahi Khameneh","email":"","orcid":"","institution":"University of Tehran","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Armin","middleName":"Jarahi","lastName":"Khameneh","suffix":""},{"id":156798229,"identity":"573b0ff4-d4c9-402b-85f5-7320f137c546","order_by":1,"name":"Ashkan Heydarian","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCUlEQVRIiWNgGAWjYFACHiBmAyPGBwwMB3igwhJEaWE2IE0LiAQqO0DYWfwNvAcf85TZ5fOxnz1W+XPPHRn+GQmMH34wWOTj0iJxgC/ZmOdcsmUbT17abZ5nz3gkbiQwS/YwSFg24NJzgMdMmreN2YCNIcfsNsOBwzwMNxIYpIFmGeDSIX+Ax/w3b1u9ARv/G7PCH0At8kBbfuPTYgC0hZm37bABm0SOGQMPUIvBjQQ2vLYYHuYxlpxz7jhQyxtjaZ4Dz3gMzzxss+wxwK1F7niP4Yc3ZdUG8v05hh9/HLhjL3c8+fCNHxV1OLUwMDMwMPGgCjE2AB2MUwNEyQ/88qNgFIyCUTDSAQBqd0whpUS27QAAAABJRU5ErkJggg==","orcid":"","institution":"Islamic Azad University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Ashkan","middleName":"","lastName":"Heydarian","suffix":""},{"id":156798230,"identity":"0f55295c-af42-4cf8-82d6-534c0f6da2e1","order_by":2,"name":"Nasibeh Babaei","email":"","orcid":"","institution":"Islamic Azad University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Nasibeh","middleName":"","lastName":"Babaei","suffix":""},{"id":156798231,"identity":"88713d7e-0707-49e8-9b01-3ac8689ed0fe","order_by":3,"name":"Negin Hannani","email":"","orcid":"","institution":"Islamic Azad University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Negin","middleName":"","lastName":"Hannani","suffix":""},{"id":156798233,"identity":"b5c116f6-a2b1-4d09-a1cc-7318e2ed1b0a","order_by":4,"name":"Dornaz Milani","email":"","orcid":"","institution":"Islamic Azad University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Dornaz","middleName":"","lastName":"Milani","suffix":""},{"id":156798235,"identity":"f179cf38-ce1b-47e1-ac11-788cc184642a","order_by":5,"name":"Hossein Sahbafar","email":"","orcid":"","institution":"University of Tehran","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hossein","middleName":"","lastName":"Sahbafar","suffix":""}],"badges":[],"createdAt":"2022-11-25 20:29:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2313700/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2313700/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":29925555,"identity":"6ef271ae-b273-4f17-92e3-473cdaa7e989","added_by":"auto","created_at":"2022-12-05 20:27:34","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":268939,"visible":true,"origin":"","legend":"\u003cp\u003eThe mean of vertical force and cell indentation for all groups.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-2313700/v1/9e67b611c7a5fef67773b240.jpeg"},{"id":29925556,"identity":"d8341064-a459-4353-944e-f98711ec18c8","added_by":"auto","created_at":"2022-12-05 20:27:35","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":31783,"visible":true,"origin":"","legend":"\u003cp\u003eThe mean and standard deviation of elastic module of all groups.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-2313700/v1/7c95c6c416a97b2c997c3cc0.png"},{"id":36200534,"identity":"28ea8b8d-a412-4c2f-9275-f00426ffe712","added_by":"auto","created_at":"2023-04-24 07:29:50","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":416463,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2313700/v1/da52b4f1-1db8-4acf-b3d3-6f8cf4c0525b.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Effects of Vitamin E and D on the Stiffness of MCF-7","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBreast cancer develops when epithelial cells multiply uncontrollably and create a mass of tissue known as a tumor. Breast cancer affects a large number of people today. Breast cancer is anticipated to be the most prevalent cancer in the United States in 2022 [1]. This disease was acknowledged globally as the leading cause of mortality among women [2].\u003c/p\u003e \u003cp\u003eThe prevention and treatment of breast cancer can benefit from using vitamins, according to investigations [3]. Both normal mammary glands and breast cancer cells have been found to express vitamin D receptors. To immunize the patient against breast cancer, vitamin D stimulates the vitamin D receptor and functions as a master transcriptional regulator of autophagy [4]. Numerous pre-clinical investigations have suggested that the 1,25D ligand plays a function in limiting normal breast gland growth and vulnerability to carcinogenesis. According to several investigations, vitamin D insufficiency is widespread in patients experiencing breast cancer, which enhances the risk of cancer development [5]. Yao et al. [6] discovered that higher levels of serum 25-hydroxyvitamin D decrease the risk of mortality and cancer among 1666 cancer patients, and this result was more significant in premenopausal women. In addition, studies evaluating the impact of vitamin D on transgenic mice models indicated that vitamin D could inhibit numerous distinctive mammary tumor models, some of which are highly comparable to human breast cancer [7\u0026ndash;13]. Both vitamin D receptor and Cyp27b1 deletion were found to promote tumorigenesis. According to research by Ooi et al. [14] osteolytic lesions manifest themselves faster in vitamin D-deficient rats than in vitamin D-sufficient rats.\u003c/p\u003e \u003cp\u003eWhen doxorubicin is combined with vitamin E, the survival of breast cancer cells is significantly shortened with minimum damage to normal cells, as demonstrated by Ahmadi et al. [15]. Despite the fact that a high dose of vitamin E exacerbates the cytotoxicity induced by doxorabicin.\u003c/p\u003e \u003cp\u003eSigounas et al. [16] discovered that vitamin E reduces cellular proliferation in breast and prostate tumor cells by lowering deoxyribonucleic acid (DNA) synthesis in cell lines. Additionally, substantial DNA fragmentation was seen while examining high molecular weight samples, suggesting that the vitamin E-supplemented cell lines underwent apoptosis.\u003c/p\u003e \u003cp\u003eNevertheless, several investigations have found that the administration of vitamin E does not affect the incidence of breast cancer [17\u0026ndash;21].\u003c/p\u003e \u003cp\u003eStiffness, nonlinearity, anisotropy, and heterogeneity are mechanical characteristics of cells. Numerous biological processes, including growth, adhesion, motility, deformation, and cell differentiation, are regulated by these characteristics [22\u0026ndash;25]. Alterations in cellular and extracellular mechanical characteristics during cancer development alter forces on cells and influence tumor cells' morphogenetic evolution, invasion, and proliferation [26, 27].\u003c/p\u003e \u003cp\u003eEvaluating the mechanical characteristics of the tumor and normal cells is one technique to differentiate them. Since the mechanical characteristics and cytoskeleton structure are altered by cell carcinogenesis [28].\u003c/p\u003e \u003cp\u003eFor instance, normal cells are less fluid [29] and more rigid than tumor cells [30\u0026ndash;32]. In addition, the stiffness of tumor cell lines tends to decrease as the severity of cancer enhances [33]. According to Plodinec et al. [34], malignant breast tissues differ from normal tissues in that they have a wider dispersion of elastic modulus and a lower stiffness peak. Tumor cells increased the contraction of the actomyosin cortex when stretched by external stimuli compared to the related normal cells [35, 36]. Additionally, there are alterations in how tumor cells interact physically with their adjacent cells and the extracellular components surrounding them, affecting the tumors' growth rate, morphology, and invasion [37, 38]. Alterations in the mechanical characteristics of cells are accompanied by alterations in the mechanical characteristics of the extracellular environment. The majority of the extracellular milieu of the tumor is made up of fibrous tissue. As breast cancer progresses, the cross-linking between these fibers grows, making the extracellular matrix environment rigid [39]. Additionally, timely detection [40], cancer prevention [41], and measuring the efficacy of anti-cancer medications can all benefit from the distinction between normal and malignant cells [31, 42].\u003c/p\u003e \u003cp\u003eAFM is one of the methods that may be employed to study cell mechanical characteristics [43]. AFM is well recognized as a useful method for investigating biological materials[44\u0026ndash;46]. It is also a versatile technique for non-destructively imaging and quantifying the elastic characteristics of living cells under physiological settings [47\u0026ndash;51]. The contact force and distance between the tip and the sample (tip-sample) are calculated using the AFM technique when the tip moves up and down at a point. Young's modulus, which provides a quantitative value of the cellular elastic characteristics, is used to represent deformation in this method. The contact-mode point-spectroscopy curve, also known as the force curve, can be used to calculate Young's modulus. Other mechanical characteristics of materials, including adhesion, deformation, and dissipation, can be determined by employing these force curves in conjunction with Young's modulus. Nevertheless, attaining these values precisely may be challenging [52]. Ex vivo mechanical characteristics of breast cancer cells, bladder cells, and fibroblasts have been studied using this method thus far [25, 53\u0026ndash;57].\u003c/p\u003e \u003cp\u003eAlthough numerous investigations have been undertaken to examine the influence of vitamins on breast cancer healing, only several investigations have been carried out to study the impact of vitamins E and D on the mechanical characteristics of breast cancer cells. This investigation was carried out in order to determine how vitamins D and E affect the mechanical characteristics of malignant breast tissues.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003eTwo cell lines, normal breast cell MCF-10 and epithelial breast cancer cell MCF-7, were obtained in two 25 ml flasks from the Stem cell Technology Research Center. The flasks were used for defreezing. The flasks were then placed inside the incubator, and after 24 hours, the cell flasks were substituted with 5 ml of Dulbecco's Modified Eagle Medium (DMEM) high Glucose culture medium comprising 10% fetal bovine serum (FBS). An inverted microscope was used to inspect the cells' density and morphology. Once they reached the appropriate density, the cells adhered to the bottom of the cell flask were detached and seeded in the suitable 3.5 cm Petri dishes. Each cell flask received 1 ml of warmed Phosphate Buffered Saline (PBS) EDTA, enough to cover the bottom of the flask. The flasks were then filled with 1 ml of warmed trypsin enzyme (37\u0026deg;C) and placed in the incubator for 2 minutes. The MCF7 cell-cell line was subsequently cultured in three 3.5 cm Petri dishes, while the MCF10 cell line was cultured in another 3.5 cm petri dish, and the Petri dishes were then placed in an incubator to achieve the necessary temperature, moisture, and CO\u003csub\u003e2\u003c/sub\u003e.\u003c/p\u003e \u003cp\u003eVitamins D3 and E were purchased in two vials from the Osvah pharmaceutical Company Vitamin D3 at 0.03 U/ml, and vitamin E at 16 U/ml concentrations were calculated and prepared to achieve the necessary volume of culture media utilized in this study. The cells were treated after 24 hours of cell culture in Petri dishes. After thoroughly removing the culture medium from the Petri dishes, 3 ml of 10% FBS culture medium comprising vitamin D3 at a concentration of 0.03 U/ml was poured into one of the Petri dishes incorporating the MCF7 cell line. Another petri dish comprising the MCF7 cell line received 3 ml of 10% FBS culture medium incorporating vitamin E at a concentration of 16U/ml. The third petri dish comprising the MCF7 cell line received 3 ml of culture medium containing 10% FBS. In addition, the supernatant in the petri dish holding the MCF10 cell line was eliminated entirely, 3 ml of 10% FBS culture medium was poured, and the dish was placed in the incubator. The appropriate four Petri dishes were brought to the laboratory for the remaining steps after receiving the treatment for 24 hours.\u003c/p\u003e \u003cp\u003eThe mechanical characteristics of the cell were evaluated using a BioAFM device manufactured by JPK brand, Germany, in this investigation. A pyramidal probe containing a square base with a pyramid apex size of 7 nm and a 35-degree angle was employed in this study. The spring constant is 0.19 Nm. The apparatus was sterilized in accordance with the protocol before initiation of the UV radiation test, and the apparatus was then calibrated. The maximum contact force of 2,700 piconewtons was exerted on 25 sites of the cells in the Petri dishes during this experiment. The force-displacement curves of 25 individual cells in each group were subsequently inspected, and the force-displacement curve was drawn to record the deformation curve. All of the test steps were conducted in 2 hours and 30 minutes at 37\u0026deg;C, and finally, the data was extracted.\u003c/p\u003e \u003cp\u003eThe Young's modulus of cell specimens was utilized to measure cell stiffness in this research, and Young's modulus was measured using the Hertz model by analyzing the data employing the MCF10, MCF7, MCF7\u0026thinsp;+\u0026thinsp;VITAMIN D3, and MCF7\u0026thinsp;+\u0026thinsp;VITAMIN E cells' force-deformation diagram.\u003c/p\u003e \u003cp\u003e \u003cspan class=\"InlineEquation\"\u003e \u003cspan class=\"mathinline\"\u003e\\(F=\\frac{E}{1-{\\nu }^{2}}\\frac{\\text{tan}\\alpha }{\\sqrt{2}}{\\delta }^{2}\\)\u003c/span\u003e \u003c/span\u003e \u003cem\u003eEq.\u0026nbsp;1\u003c/em\u003e\u003c/p\u003e \u003cp\u003eIn the probe formula, α represents the half angle to the probe's face (55 \u0026deg;), ϑ represents Poisson's ratio in δ-Phase, which is believed to be 0.5 for soft biological samples and denotes the deformation depth, F indicates the force on the cell, and E represents Young's modulus. Each cell's Young's modulus was determined. The data from four groups were averaged, and two groups treated with vitamin D and E were compared to MCF-10 and MCF-7 control groups. The normality test was employed to determine the non-randomness of the data collected from the Mini Tab software. The one-way ANOVA test was used to analyze the average of the four groups.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe cells in each group's deformation were analyzed. Initially, the console's force and the AFM tip displacement were both assumed to be zero. Subsequently, for each cell in each group, the force-indentation data were analyzed, and the mean force-indentation data for all four groups were drawn (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eEach group's loading and unloading processes are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\n\u003cp\u003eMCF-10 cells, MCF-7 tumor cells, MCF-7 tumor cells treated with vitamin D, and MCF-7 tumor cells treated with vitamin E had maximum membrane deformations of 7.318, 0.293, 7.821 and 0.382 \u0026micro;m, respectively.\u003c/p\u003e\n\u003cp\u003eYoung's modulus and the Hertz model were determined for all four cell lines. MCF10 cells, MCF7 treated with vitamin D, MCF7 treated with vitamin E, and MCF7 had mean values and SD standard deviations of 455.6\u0026thinsp;\u0026plusmn;\u0026thinsp;175.7, 585.8\u0026thinsp;\u0026plusmn;\u0026thinsp;138, 216.6\u0026thinsp;\u0026plusmn;\u0026thinsp;40.82, and 203.4\u0026thinsp;\u0026plusmn;\u0026thinsp;43.1 Pa, respectively (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eANOVA test with a 95% confidence interval was used to compare the averages of the four groups and determine their difference. A statistically significant difference was found in the average of these groups (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\n\u003cp\u003eTukey's test for post-Hoc analysis was subsequently performed to assess the statistical significance and pairwise comparison of cell groups. Young's modulus was shown to differ across MCF10, MCF-7, MCF7\u0026thinsp;+\u0026thinsp;E, and MCF7\u0026thinsp;+\u0026thinsp;D3 groups.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe mechanical characteristics of normal breast epithelial cells were compared with cancer cells treated with vitamin D and E in this research to evaluate the short-term (for 24 hours) impact of vitamins D and E on the mechanical properties of tumor cells. The Young's modulus acquired from the Hertz model of tumor cells treated with vitamin D was substantially similar to Young's modulus of normal cells, according to the results of this research. Nevertheless, supplementing vitamin E did not affect the characteristics of tumor cells, and Young's modulus stayed nearer to cancer cells rather than normal cells.\u003c/p\u003e \u003cp\u003eTumor cells become more rigid as a result of vitamin D supplementation, approaching the normal cells' stiffness. According to research, normal cells are stiffer than malignant cells [30, 32, 58]. The stiffness of the cells decreases as the malignancy progresses [19]. According to Li et al. [58, 59], decreased stiffness and a notable reduction of F-actin filaments or their bundles (stress fibers) may be the cause of the cancer cells' easier motility and invasion during metastasis. As a result, by stiffening tumor cells, vitamin D can limit metastasis and migration.\u003c/p\u003e \u003cp\u003eThe mechanical characteristics of the cells were unaffected by vitamin E treatment. There was no correlation between vitamin E and a lower risk of breast cancer in other investigations [18, 19]. Hunter et al. [20] also reported that high vitamin E supplementation did not protect women from breast cancer.\u003c/p\u003e \u003cp\u003eBecause vitamin E is lipophilic [60], various investigations have revealed that it has the potential to affect the biophysical characteristics of membrane and lipid species. Asif et al. [61] discovered that administering vitamin E as an anti-tumor medication induced substantial DNA damage, mitochondrial depolarization, apoptosis, and arrest of tumor cell proliferation. Additionally, it has been shown that treatment with TC6OH, a vitamin E derivative, induces tumor cell apoptosis by transport, breakdown, and lowering of some anti-apoptotic and growth factor-related proteins, damaging the mitochondrial membrane potential and modulating lipids [62].\u003c/p\u003e \u003cp\u003eFurther research is proposed to evaluate the effect of the vitamins D, and E on tumor cells and that tumor cells are treated with varying doses of these vitamins to achieve more precise and reliable findings. This can respond to the question of how many doses of these vitamins can be more effective or even detrimental. It is important to note that the long-term impact of these vitamins on tumor cells is a topic that can be discussed in the future.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn summary, this research demonstrates that despite vitamin E, vitamin D has a major impact on altering the mechanical characteristics (stiffness) of tumor cells and can make them closer to the normal cells' properties, suggesting that vitamin D may be helpful in the treatment of cancer. It can also decrease the possibility of cell migration by enhancing cell stiffness.\u003c/p\u003e"},{"header":"List Of Abbreviations","content":"\u003cp\u003e\u003cstrong\u003eAbbreviation\u003c/strong\u003e \u003cstrong\u003eDefinition\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAFM Atomic force microscopy\u003c/p\u003e\n\u003cp\u003eDNA Deoxyribonucleic acid\u003c/p\u003e\n\u003cp\u003e(DMEM) Dulbecco's Modified Eagle Medium\u003c/p\u003e\n\u003cp\u003eFBS Fetal Bovine Serum\u003c/p\u003e\n\u003cp\u003ePBS Phosphate Buffered Saline\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. This article does not contain any studies with human participants or animals performed by any of the authors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors consent for publication.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors of the manuscript hereby declare that they have no conflict of interest\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere was no funding included.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eArmin Jarahi, Ashkan Heydarian and Dornaz Millani were performed the experimental testing, modeling and statistical analysis Negin Hannani and Nasibe Babaei wrote the manuscript. Hosein Sahbafar were performed cell culturing.All authors reviewed the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSiegel, R. L., Miller, K. D., Fuchs, H. E., \u0026amp; Jemal, A. (2022). Cancer statistics, 2022. \u003cem\u003eCA: a cancer journal for clinicians\u003c/em\u003e, \u003cem\u003e72\u003c/em\u003e(1), 7\u0026ndash;33. https://doi.org/10.3322/CAAC.21708\u003c/li\u003e\n\u003cli\u003eSung, H., Ferlay, J., Siegel, R. L., Laversanne, M., Soerjomataram, I., Jemal, A., \u0026amp; Bray, F. (2021). Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. \u003cem\u003eCA: A Cancer Journal for Clinicians\u003c/em\u003e, \u003cem\u003e71\u003c/em\u003e(3), 209\u0026ndash;249. https://doi.org/10.3322/CAAC.21660\u003c/li\u003e\n\u003cli\u003eSantos, J. M., Khan, Z. S., Munir, M. T., Tarafdar, K., Rahman, S. M., \u0026amp; Hussain, F. (2018). Vitamin D3 decreases glycolysis and invasiveness, and increases cellular stiffness in breast cancer cells. \u003cem\u003eThe Journal of Nutritional Biochemistry\u003c/em\u003e, \u003cem\u003e53\u003c/em\u003e, 111\u0026ndash;120. https://doi.org/10.1016/J.JNUTBIO.2017.10.013\u003c/li\u003e\n\u003cli\u003eTavera-Mendoza, L. E., Westerling, T., Libby, E., Marusyk, A., Cato, L., Cassani, R., \u0026hellip; Brown, M. (2017). Vitamin D receptor regulates autophagy in the normal mammary gland and in luminal breast cancer cells. \u003cem\u003eProceedings of the National Academy of Sciences of the United States of America\u003c/em\u003e, \u003cem\u003e114\u003c/em\u003e(11), E2186\u0026ndash;E2194. https://doi.org/10.1073/PNAS.1615015114\u003c/li\u003e\n\u003cli\u003emolecular, J. W.-T. J. of steroid biochemistry and, \u0026amp; 2018, undefined. (n.d.). Vitamin D and breast cancer: Past and present. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S0960076017301905\u003c/li\u003e\n\u003cli\u003eYao, S., Kwan, M. L., Ergas, I. J., Roh, J. M., Cheng, T. Y. D., Hong, C. C., \u0026hellip; Kushi, L. H. (2017). Association of Serum Level of Vitamin D at Diagnosis With Breast Cancer Survival: A Case-Cohort Analysis in the Pathways Study. \u003cem\u003eJAMA Oncology\u003c/em\u003e, \u003cem\u003e3\u003c/em\u003e(3), 351\u0026ndash;357. https://doi.org/10.1001/JAMAONCOL.2016.4188\u003c/li\u003e\n\u003cli\u003eSo, J. Y., Wahler, J. E., Yoon, T., Smolarek, A. K., Lin, Y., Shih, W. J., \u0026hellip; Suh, N. (2013). Oral administration of a Gemini vitamin D analog, a synthetic triterpenoid and the combination prevents mammary tumorigenesis driven by ErbB2 overexpression. \u003cem\u003eCancer Prevention Research\u003c/em\u003e, \u003cem\u003e6\u003c/em\u003e(9), 959\u0026ndash;970. https://doi.org/10.1158/1940-6207.CAPR-13-0087/35984/AM/ORAL-ADMINISTRATION-OF-A-GEMINI-VITAMIN-D-ANALOG-A\u003c/li\u003e\n\u003cli\u003eMilliken, E. L., Zhang, X., Flask, C., Duerk, J. L., MacDonald, P. N., \u0026amp; Keri, R. A. (2005). EB1089, a vitamin D receptor agonist, reduces proliferation and decreases tumor growth rate in a mouse model of hormone-induced mammary cancer. \u003cem\u003eCancer Letters\u003c/em\u003e, \u003cem\u003e229\u003c/em\u003e(2), 205\u0026ndash;215. https://doi.org/10.1016/J.CANLET.2005.06.044\u003c/li\u003e\n\u003cli\u003eLee, H. J., So, J. Y., DeCastro, A., Smolarek, A., Paul, S., Maehr, H., \u0026hellip; Suh, N. (2010). Gemini vitamin D analog suppresses ErbB2-positive mammary tumor growth via inhibition of ErbB2/AKT/ERK signaling. \u003cem\u003eThe Journal of Steroid Biochemistry and Molecular Biology\u003c/em\u003e, \u003cem\u003e121\u003c/em\u003e(1\u0026ndash;2), 408\u0026ndash;412. https://doi.org/10.1016/J.JSBMB.2010.03.053\u003c/li\u003e\n\u003cli\u003eZinser, G. M., \u0026amp; Welsch, J. E. (2004). Vitamin D receptor status alters mammary gland morphology and tumorigenesis in MMTV-neu mice. \u003cem\u003eCarcinogenesis\u003c/em\u003e, \u003cem\u003e25\u003c/em\u003e(12), 2361\u0026ndash;2372. https://doi.org/10.1093/CARCIN/BGH271\u003c/li\u003e\n\u003cli\u003eLi, J., Luco, A. L., Ochietti, B., Fadhil, I., Camirand, A., Reinhardt, T. A., \u0026hellip; Kremer, R. (2016). Tumoral Vitamin D Synthesis by CYP27B1 1-\u0026alpha;-Hydroxylase Delays Mammary Tumor Progression in the PyMT-MMTV Mouse Model and Its Action Involves NF-\u0026kappa;B Modulation. \u003cem\u003eEndocrinology\u003c/em\u003e, \u003cem\u003e157\u003c/em\u003e(6), 2204\u0026ndash;2216. https://doi.org/10.1210/EN.2015-1824\u003c/li\u003e\n\u003cli\u003eLi, J., Luco, A. L., Huang, D. C., Reinhardt, T. A., Muller, W., \u0026amp; Kremer, R. (2015). Chemoprevention activity of 25-hydroxyvitamin d in the MMTV-PyMT mouse model of breast cancer. \u003cem\u003eCancer Prevention Research\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(2), 120\u0026ndash;128. https://doi.org/10.1158/1940-6207.CAPR-14-0110/36341/AM/CHEMOPREVENTION-ACTIVITY-OF-25-HYDROXYVITAMIN-D-IN\u003c/li\u003e\n\u003cli\u003eJohnson, A. L., Zinser, G. M., \u0026amp; Waltz, S. E. (2015). Vitamin D3-dependent VDR signaling delays ron-mediated breast tumorigenesis through suppression of \u0026beta;-catenin activity. \u003cem\u003eOncotarget\u003c/em\u003e, \u003cem\u003e6\u003c/em\u003e(18), 16304. https://doi.org/10.18632/ONCOTARGET.4059\u003c/li\u003e\n\u003cli\u003eOoi, L. L., Zhou, H., Kalak, R., Zheng, Y., Conigrave, A. D., Seibel, M. J., \u0026amp; Dunstan, C. R. (2010). Vitamin D deficiency promotes human breast cancer growth in a murine model of bone metastasis. \u003cem\u003eCancer Research\u003c/em\u003e, \u003cem\u003e70\u003c/em\u003e(5), 1835\u0026ndash;1844. https://doi.org/10.1158/0008-5472.CAN-09-3194/655774/P/VITAMIN-D-DEFICIENCY-PROMOTES-HUMAN-BREAST-CANCER\u003c/li\u003e\n\u003cli\u003eAhmadi, M., Hedayatizadeh-Omran, A., Alizadeh-Navaei, R., Saeedi, M., Zaboli, E., Amjadi, O., \u0026hellip; Besharat, Z. (2022). Effects of Vitamin E on Doxorubicin Cytotoxicity in Human Breast Cancer Cells in Vitro. \u003cem\u003eAsian Pacific Journal of Cancer Prevention\u003c/em\u003e, \u003cem\u003e23\u003c/em\u003e(1), 201\u0026ndash;205. https://doi.org/10.31557/APJCP.2022.23.1.201\u003c/li\u003e\n\u003cli\u003eSigounas, G., Anagnostou, A., \u0026amp; Steiner, M. (2009). dl‐\u0026alpha;‐tocopherol induces apoptosis in erythroleukemia, prostate, and breast cancer cells. \u003cem\u003ehttp://dx.doi.org/10.1080/01635589709514549\u003c/em\u003e, \u003cem\u003e28\u003c/em\u003e(1), 30\u0026ndash;35. https://doi.org/10.1080/01635589709514549\u003c/li\u003e\n\u003cli\u003eSato, R., Helzlsouer, K., Alberg, A., \u0026hellip; S. H.-C. E., \u0026amp; 2002, undefined. (2002). Prospective study of carotenoids, tocopherols, and retinoid concentrations and the risk of breast cancer. \u003cem\u003eAACR\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e, 451\u0026ndash;457. Retrieved from https://aacrjournals.org/cebp/article-abstract/11/5/451/166599\u003c/li\u003e\n\u003cli\u003eKushi, L., Fee, R., Sellers, T., \u0026hellip; W. Z.-A. journal of, \u0026amp; 1996, undefined. (1996). Intake of vitamins A, C, and E and postmenopausal breast cancer: the Iowa Women\u0026rsquo;s Health Study. \u003cem\u003eacademic.oup.com\u003c/em\u003e, \u003cem\u003e144\u003c/em\u003e(2). Retrieved from https://academic.oup.com/aje/article-abstract/144/2/165/112184\u003c/li\u003e\n\u003cli\u003eRohan, T. E., Howe, G. R., Friedenreich, C. M., Jain, M., \u0026amp; Miller, A. B. (1993). Dietary fiber, vitamins A, C, and E, and risk of breast cancer: a cohort study. \u003cem\u003eCancer Causes and Control\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e(1), 29\u0026ndash;37. https://doi.org/10.1007/BF00051711\u003c/li\u003e\n\u003cli\u003eHunter, D. J., Manson, J. E., Colditz, G. A., Stampfer, M. J., Rosner, B., Hennekens, C. H., \u0026hellip; Willett, W. C. (1993). A Prospective Study of the Intake of Vitamins C, E, and A and the Risk of Breast Cancer. \u003cem\u003eNew England Journal of Medicine\u003c/em\u003e, \u003cem\u003e329\u003c/em\u003e(4), 234\u0026ndash;240. https://doi.org/10.1056/NEJM199307223290403\u003c/li\u003e\n\u003cli\u003eZhang, S., Hunter, D., \u0026hellip; M. F.-J. of the, \u0026amp; 1999, undefined. (n.d.). Dietary carotenoids and vitamins A, C, and E and risk of breast cancer. \u003cem\u003eacademic.oup.com\u003c/em\u003e. Retrieved from https://academic.oup.com/jnci/article-abstract/91/6/547/2549364\u003c/li\u003e\n\u003cli\u003eZhu, C., Bao, G., engineering, N. W.-A. review of biomedical, \u0026amp; 2000, undefined. (n.d.). Cell mechanics: mechanical response, cell adhesion, and molecular deformation. \u003cem\u003eresearchgate.net\u003c/em\u003e. Retrieved from https://www.researchgate.net/profile/Cheng-Zhu-32/publication/11652210_C_ELL_M_ECHANICS_Mechanical_Response_Cell_Adhesion_and_Molecular_Deformation/links/53da1d8f0cf2631430c7f843/C-ELL-M-ECHANICS-Mechanical-Response-Cell-Adhesion-and-Molecular-Deformation.pdf?_sg%5B0%5D=started_experiment_milestone\u0026amp;origin=journalDetail\u003c/li\u003e\n\u003cli\u003echemistry, E. E.-A. review of biophysics and biophysical, \u0026amp; 1988, undefined. (1988). Cellular mechanics as an indicator of cytoskeletal structure and function. \u003cem\u003eresearchgate.net\u003c/em\u003e. https://doi.org/10.1146/annurev.bb.17.060188.002145\u003c/li\u003e\n\u003cli\u003ePourati, J., Maniotis, A., Spiegel, D., Schaffer, J. L., Butler, J. P., Fredberg, J. J., \u0026hellip; Wang, N. (1998). Is cytoskeletal tension a major determinant of cell deformability in adherent endothelial cells? \u003cem\u003eAmerican Journal of Physiology - Cell Physiology\u003c/em\u003e, \u003cem\u003e274\u003c/em\u003e(5 43-5). https://doi.org/10.1152/AJPCELL.1998.274.5.C1283\u003c/li\u003e\n\u003cli\u003eRotsch, C., Jacobson, K., \u0026amp; Radmacher, M. (1999). Dimensional and mechanical dynamics of active and stable edges in motile fibroblasts investigated by using atomic force microscopy. \u003cem\u003eProceedings of the National Academy of Sciences of the United States of America\u003c/em\u003e, \u003cem\u003e96\u003c/em\u003e(3), 921\u0026ndash;926. https://doi.org/10.1073/PNAS.96.3.921\u003c/li\u003e\n\u003cli\u003eHuang, S., cell, D. I.-C., \u0026amp; 2005, undefined. (n.d.). Cell tension, matrix mechanics, and cancer development. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S1535610805002679\u003c/li\u003e\n\u003cli\u003eLopez, J. I., Mouw, J. K., \u0026amp; Weaver, V. M. (2008). Biomechanical regulation of cell orientation and fate. \u003cem\u003eOncogene 2008 27:55\u003c/em\u003e, \u003cem\u003e27\u003c/em\u003e(55), 6981\u0026ndash;6993. https://doi.org/10.1038/onc.2008.348\u003c/li\u003e\n\u003cli\u003eSuresh, S., Spatz, J., Mills, J. P., Micoulet, A., Dao, M., Lim, C. T., \u0026hellip; Seufferlein, T. (2005). Connections between single-cell biomechanics and human disease states: gastrointestinal cancer and malaria. \u003cem\u003eActa Biomaterialia\u003c/em\u003e, \u003cem\u003e1\u003c/em\u003e(1), 15\u0026ndash;30. https://doi.org/10.1016/J.ACTBIO.2004.09.001\u003c/li\u003e\n\u003cli\u003eRother, J., N\u0026ouml;ding, H., Mey, I., \u0026amp; Janshoff, A. (2014). Atomic force microscopy-based microrheology reveals significant differences in the viscoelastic response between malign and benign cell lines. \u003cem\u003eOpen Biology\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e(MAY). https://doi.org/10.1098/RSOB.140046\u003c/li\u003e\n\u003cli\u003eCoughlin, M. F., Bielenberg, D. R., Lenormand, G., Marinkovic, M., Waghorne, C. G., Zetter, B. R., \u0026amp; Fredberg, J. J. (2012). Cytoskeletal stiffness, friction, and fluidity of cancer cell lines with different metastatic potential. \u003cem\u003eClinical \u0026amp; Experimental Metastasis 2012 30:3\u003c/em\u003e, \u003cem\u003e30\u003c/em\u003e(3), 237\u0026ndash;250. https://doi.org/10.1007/S10585-012-9531-Z\u003c/li\u003e\n\u003cli\u003eGuck, J., Schinkinger, S., Lincoln, B., Wottawah, F., journal, S. E.-B., \u0026amp; 2005, undefined. (n.d.). Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349505734172\u003c/li\u003e\n\u003cli\u003eSwaminathan, V., Mythreye, K., Tim O\u0026rsquo;Brien, E., Berchuck, A., Blobe, G. C., \u0026amp; Superfine, R. (2011). Mechanical Stiffness grades metastatic potential in patient tumor cells and in cancer cell lines. \u003cem\u003eCancer Research\u003c/em\u003e, \u003cem\u003e71\u003c/em\u003e(15), 5075\u0026ndash;5080. https://doi.org/10.1158/0008-5472.CAN-11-0247/649738/AM/MECHANICAL-STIFFNESS-GRADES-METASTATIC-POTENTIAL\u003c/li\u003e\n\u003cli\u003eSuresh, S. (2007). Biomechanics and biophysics of cancer cells. \u003cem\u003eActa Biomaterialia\u003c/em\u003e, \u003cem\u003e3\u003c/em\u003e(4), 413\u0026ndash;438. https://doi.org/10.1016/J.ACTBIO.2007.04.002\u003c/li\u003e\n\u003cli\u003ePlodinec, M., Loparic, M., Monnier, C. A., Obermann, E. C., Zanetti-Dallenbach, R., Oertle, P., \u0026hellip; Schoenenberger, C. A. (2012). The nanomechanical signature of breast cancer. \u003cem\u003eNature Nanotechnology 2012 7:11\u003c/em\u003e, \u003cem\u003e7\u003c/em\u003e(11), 757\u0026ndash;765. https://doi.org/10.1038/nnano.2012.167\u003c/li\u003e\n\u003cli\u003eJonas, O., Mierke, C. T., \u0026amp; K\u0026auml;s, J. A. (2011). Invasive cancer cell lines exhibit biomechanical properties that are distinct from their noninvasive counterparts. \u003cem\u003eSoft Matter\u003c/em\u003e, \u003cem\u003e7\u003c/em\u003e(24), 11488\u0026ndash;11495. https://doi.org/10.1039/C1SM05532A\u003c/li\u003e\n\u003cli\u003eKraning-Rush, C. M., Califano, J. P., \u0026amp; Reinhart-King, C. A. (2012). Cellular Traction Stresses Increase with Increasing Metastatic Potential. \u003cem\u003ePLOS ONE\u003c/em\u003e, \u003cem\u003e7\u003c/em\u003e(2), e32572. https://doi.org/10.1371/JOURNAL.PONE.0032572\u003c/li\u003e\n\u003cli\u003eParedes, J., Albergaria, A., Oliveira, J. T., Jeronimo, C., Milanezi, F., \u0026amp; Schmitt, F. C. (2005). P-Cadherin Overexpression Is an Indicator of Clinical Outcome in Invasive Breast Carcinomas and Is Associated with CDH3 Promoter Hypomethylation. \u003cem\u003eClinical Cancer Research\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e(16), 5869\u0026ndash;5877. https://doi.org/10.1158/1078-0432.CCR-05-0059\u003c/li\u003e\n\u003cli\u003eRibeiro, A. S., Albergaria, A., Sousa, B., Correia, A. L., Bracke, M., Seruca, R., \u0026hellip; Paredes, J. (2009). Extracellular cleavage and shedding of P-cadherin: a mechanism underlying the invasive behaviour of breast cancer cells. \u003cem\u003eOncogene 2010 29:3\u003c/em\u003e, \u003cem\u003e29\u003c/em\u003e(3), 392\u0026ndash;402. https://doi.org/10.1038/onc.2009.338\u003c/li\u003e\n\u003cli\u003eAnsardamavandi, A., Tafazzoli-Shadpour, M., Omidvar, R., \u0026amp; Jahanzad, I. (2016). Quantification of effects of cancer on elastic properties of breast tissue by Atomic Force Microscopy. \u003cem\u003eJournal of the Mechanical Behavior of Biomedical Materials\u003c/em\u003e, \u003cem\u003e60\u003c/em\u003e, 234\u0026ndash;242. https://doi.org/10.1016/J.JMBBM.2015.12.028\u003c/li\u003e\n\u003cli\u003eLee, G., biotechnology, C. L.-T. in, \u0026amp; 2007, undefined. (n.d.). Biomechanics approaches to studying human diseases. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S0167779907000212\u003c/li\u003e\n\u003cli\u003eKatira, P., Bonnecaze, R. T., \u0026amp; Zaman, M. H. (2013). Modeling the mechanics of cancer: Effect of changes in cellular and extra-cellular mechanical properties. \u003cem\u003eFrontiers in Oncology\u003c/em\u003e, \u003cem\u003e3 JUN\u003c/em\u003e. https://doi.org/10.3389/FONC.2013.00145/FULL\u003c/li\u003e\n\u003cli\u003eWottawah, F., Schinkinger, S., Lincoln, B., biomaterialia, S. E.-A., \u0026amp; 2005, undefined. (n.d.). Characterizing single suspended cells by optorheology. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S1742706105000589\u003c/li\u003e\n\u003cli\u003eBinnig, G., Quate, C. F., \u0026amp; Gerber, C. (1986). Atomic force microscope. \u003cem\u003ePhysical Review Letters\u003c/em\u003e, \u003cem\u003e56\u003c/em\u003e(9), 930\u0026ndash;933. https://doi.org/10.1103/PHYSREVLETT.56.930\u003c/li\u003e\n\u003cli\u003eKasas, S., Thomson, N. H., Smith, B. L., Hansma, P. K., Miklossy, J., \u0026amp; Hansma, H. G. (1996). Biological Applications of the AFM: From Single Molecules to Organs. https://doi.org/10.1002/(SICI)1098-1098(1997)8:2\u003c/li\u003e\n\u003cli\u003escience, E. H.-P. in surface, \u0026amp; 1994, undefined. (n.d.). Imaging of living cells by atomic force microscopy. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/007968169490006X\u003c/li\u003e\n\u003cli\u003eHansma, H., and, J. H.-A. review of biophysics, \u0026amp; 1994, undefined. (n.d.). Biomolecular imaging with the atomic force microscope. \u003cem\u003ejhu.pure.elsevier.com\u003c/em\u003e. Retrieved from https://jhu.pure.elsevier.com/en/publications/biomolecular-imaging-with-the-atomic-force-microscope-4\u003c/li\u003e\n\u003cli\u003eSchoenenberger, C., journal, J. H.-B., \u0026amp; 1994, undefined. (n.d.). Slow cellular dynamics in MDCK and R5 cells monitored by time-lapse atomic force microscopy. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349594805569\u003c/li\u003e\n\u003cli\u003eA-Hassan, E., Heinz, W. F., Antonik, M. D., D\u0026rsquo;Costa, N. P., Nageswaran, S., Schoenenberger, C. A., \u0026amp; Hoh, J. H. (1998). Relative Microelastic Mapping of Living Cells by Atomic Force Microscopy. \u003cem\u003eBiophysical Journal\u003c/em\u003e, \u003cem\u003e74\u003c/em\u003e(3), 1564\u0026ndash;1578. https://doi.org/10.1016/S0006-3495(98)77868-3\u003c/li\u003e\n\u003cli\u003eSchaus, S., journal, E. H.-B., \u0026amp; 1997, undefined. (n.d.). Cell viability and probe-cell membrane interactions of XR1 glial cells imaged by atomic force microscopy. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349597781530\u003c/li\u003e\n\u003cli\u003eHaydon, P. G., Lartius, R., Parpura, V., \u0026amp; Marchese-Ragona, S. P. (1996). Membrane deformation of living glial cells using atomic force microscopy. \u003cem\u003eJournal of Microscopy\u003c/em\u003e, \u003cem\u003e182\u003c/em\u003e(2), 114\u0026ndash;120. https://doi.org/10.1046/J.1365-2818.1996.141423.X\u003c/li\u003e\n\u003cli\u003eHenderson, E., Haydon, P. G., \u0026amp; Sakaguchi, D. S. (1992). Actin filament dynamics in living glial cells imaged by atomic force microscopy. \u003cem\u003eScience\u003c/em\u003e, \u003cem\u003e257\u003c/em\u003e(5078), 1944\u0026ndash;1946. https://doi.org/10.1126/SCIENCE.1411511\u003c/li\u003e\n\u003cli\u003ePolytech Elec Eng Comp Sci K\u0026aacute;m\u0026aacute;n, P. J., \u0026amp; K\u0026aacute;m\u0026aacute;n, J. (2015). Young\u0026rsquo;s Modulus and Energy Dissipation Determination Methods by AFM, with Particular Reference to a Chalcogenide Thin Film. \u003cem\u003ePeriodica Polytechnica Electrical Engineering and Computer Science\u003c/em\u003e, \u003cem\u003e59\u003c/em\u003e(1), 18\u0026ndash;25. https://doi.org/10.3311/PPEE.7865\u003c/li\u003e\n\u003cli\u003eLekka, M., Laidler, P., Ignacak, J., Łabędź, M., \u0026hellip; J. L.-\u0026hellip; et B. A. (BBA, \u0026amp; 2001, undefined. (n.d.). The effect of chitosan on stiffness and glycolytic activity of human bladder cells. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S0167488901001252\u003c/li\u003e\n\u003cli\u003eRotsch, C., journal, M. R.-B., \u0026amp; 2000, undefined. (n.d.). Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts: an atomic force microscopy study. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349500766148\u003c/li\u003e\n\u003cli\u003eLekka, M., Laidler, P., Gil, D., Lekki, J., \u0026hellip; Z. S.-E. B., \u0026amp; 1999, undefined. (1999). Elasticity of normal and cancerous human bladder cells studied by scanning force microscopy. \u003cem\u003eSpringer\u003c/em\u003e, \u003cem\u003e28\u003c/em\u003e(4), 312\u0026ndash;316. https://doi.org/10.1007/s002490050213\u003c/li\u003e\n\u003cli\u003ePark, S., Koch, D., Cardenas, R., K\u0026auml;s, J., journal, C. S.-B., \u0026amp; 2005, undefined. (n.d.). Cell motility and local viscoelasticity of fibroblasts. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S0006349505730733\u003c/li\u003e\n\u003cli\u003eCross, S. E., Jin, Y.-S., Rao, J., \u0026amp; Gimzewski, J. K. (2020). Nanomechanical Analysis of Cells from Cancer Patients. \u003cem\u003eNano-Enabled Medical Applications\u003c/em\u003e, 547\u0026ndash;566. https://doi.org/10.1201/9780429399039-18/NANOMECHANICAL-ANALYSIS-CELLS-CANCER-PATIENTS-SARAH-CROSS-YU-SHENG-JIN-JIANYU-RAO-JAMES-GIMZEWSKI\u003c/li\u003e\n\u003cli\u003eLi, Q. S., Lee, G. Y. H., Ong, C. N., \u0026amp; Lim, C. T. (2009). Probing the Elasticity of Breast Cancer Cells Using AFM. \u003cem\u003eIFMBE Proceedings\u003c/em\u003e, \u003cem\u003e23\u003c/em\u003e, 2122\u0026ndash;2125. https://doi.org/10.1007/978-3-540-92841-6_530\u003c/li\u003e\n\u003cli\u003eLi, Q. S., Lee, G. Y. H., Ong, C. N., \u0026amp; Lim, C. T. (2008). AFM indentation study of breast cancer cells. \u003cem\u003eBiochemical and Biophysical Research Communications\u003c/em\u003e, \u003cem\u003e374\u003c/em\u003e(4), 609\u0026ndash;613. https://doi.org/10.1016/J.BBRC.2008.07.078\u003c/li\u003e\n\u003cli\u003eWang, X., \u0026amp; Quinn, P. J. (2000). The location and function of vitamin E in membranes (review). \u003cem\u003eMolecular membrane biology\u003c/em\u003e, \u003cem\u003e17\u003c/em\u003e(3), 143\u0026ndash;156. https://doi.org/10.1080/09687680010000311\u003c/li\u003e\n\u003cli\u003eItoo, A., Paul, M., Ghosh, B., Polymers, S. B.-C., \u0026amp; 2022, undefined. (n.d.). Oxaliplatin delivery via chitosan/vitamin E conjugate micelles for improved efficacy and MDR-reversal in breast cancer. \u003cem\u003eElsevier\u003c/em\u003e. Retrieved from https://www.sciencedirect.com/science/article/pii/S0144861722000121\u003c/li\u003e\n\u003cli\u003eGok, S., Kuzmenko, O., Babinskyi, A., \u0026amp; Severcan, F. (2021). Vitamin E Derivative with Modified Side Chain Induced Apoptosis by Modulating the Cellular Lipids and Membrane Dynamics in MCF7 Cells. \u003cem\u003eCell Biochemistry and Biophysics 2021 79:2\u003c/em\u003e, \u003cem\u003e79\u003c/em\u003e(2), 271\u0026ndash;287. https://doi.org/10.1007/S12013-020-00961-Y\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Breast Cancer, Vitamin D, Vitamin E, Elasticity, Epithelial Cell, Cell Mechanics","lastPublishedDoi":"10.21203/rs.3.rs-2313700/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2313700/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eSeveral investigations have demonstrated that vitamins can be used to treat or prevent cancer by altering actin filaments, inhibiting cell migration and cancer cell proliferation. Vitamins D and E are fat-soluble. This research aims to determine the short-term impact of vitamin D and E on the mechanical characteristics of breast cancer cells before comparing them with normal breast cells.\u003c/p\u003e\u003ch2\u003eMethod\u003c/h2\u003e \u003cp\u003eAtomic force microscopy (AFM) was used to examine the deformation of MCF-10 normal breast cells, MCF-7 breast cancer cells, and MCF-7 breast cancer cells treated with 0.03 \u0026micro;M vitamin D and 16 \u0026micro;M vitamin E solution. Young's modulus was calculated employing the Hertz model to determine cell stiffness.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThe Young's modulus of vitamin D-treated cancer cells (585.8 Pa) was substantially similar to that of normal cells (455.6 Pa). Nevertheless, vitamin E treatment had no effect on Young's modulus of cancer cells, which remained remarkably similar to that of untreated cancer cells (216.6 and 203.4 Pa, respectively).\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eDespite vitamin E, vitamin D enhances the stiffness of tumor cells and puts their mechanical characteristics similar to normal cells by interfering with actin filaments and cell skeletons, which may inhibit tumor cell migration. According to these findings, Vitamin D appears to be an effective drug for cancer treatment.\u003c/p\u003e","manuscriptTitle":"Effects of Vitamin E and D on the Stiffness of MCF-7","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-12-05 20:27:30","doi":"10.21203/rs.3.rs-2313700/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"dfbecc98-3af8-4f2f-8445-a15fe8bfe3fb","owner":[],"postedDate":"December 5th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-04-24T07:29:33+00:00","versionOfRecord":[],"versionCreatedAt":"2022-12-05 20:27:30","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2313700","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2313700","identity":"rs-2313700","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}