Comparative Analysis of Inflammatory Biomarkers and Lymphocyte Levels in Breast Cancer Patients and Healthy Controls

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Inflammation-related hematologic parameters—such as the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR)—have been proposed as potential biomarkers for cancer progression. However, data on their association with breast cancer in North Macedonia are limited. Aim: This study aimed to evaluate NLR, PLR, and LMR in patients with breast cancer compared to healthy controls, to explore their potential as inflammatory biomarkers for early detection. Methods: A case-control study was conducted in 2025 at the Department of Medical Biochemistry, Clinical Hospital "Dr. Trifun Panovski" in Bitola. Sixty-two women were enrolled -31 with histologically confirmed breast cancer and 31 age-matched healthy controls. Hematological parameters were measured using a Sysmex XP-300 hematology analyzer. Statistical analyses were performed using SPSS v13.0. Results: The mean lymphocyte count was significantly higher in breast cancer patients compared to controls (p = 0.041). No statistically significant differences were found in PLR (p = 0.087), NLR (p = 0.477), or LMR (p = 0.141) between groups. Hemoglobin levels were significantly lower in the breast cancer group (p = 0.001). Conclusion: While lymphocyte levels and hemoglobin were significantly altered in breast cancer patients, our results did not support NLR, PLR, or LMR as standalone markers for early breast cancer detection in this population. Further large-scale studies are needed to validate these findings. breast cancer NLR PLR LMR inflammation biomarkers hematology early detection Figures Figure 1 Figure 2 Figure 3 Introduction Breast cancer (BC) remains the most frequently diagnosed malignancy and the leading cause of cancer-related death among women worldwide, accounting for approximately 2.3 million new cases and 685,000 deaths in 2020 [1]. Despite advances in diagnosis and treatment, a significant proportion of patients still experience recurrence or distant metastases within five years of initial diagnosis [2,3]. Systemic inflammation has been increasingly recognized as a critical component in the pathogenesis and progression of cancer, including BC [4–6]. Tumor cells can provoke systemic inflammatory responses, leading to measurable changes in hematologic parameters such as neutrophils, lymphocytes, platelets, and monocytes [7]. These inflammatory changes may be reflected in specific ratios derived from complete blood counts—namely the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR)—which have been investigated as potential diagnostic and prognostic biomarkers in several solid tumors, including breast cancer [8–10]. These markers are inexpensive, routinely available, and non-invasive, making them especially valuable in low- and middle-income countries where access to advanced diagnostic tools may be limited [11,12]. While several studies have supported the utility of NLR, PLR, and LMR in the prognosis of BC, findings have been inconsistent, particularly regarding their diagnostic value [13–15]. In North Macedonia, data on the association between these hematological indices and breast cancer are scarce. Therefore, the aim of this study was to evaluate the levels of NLR, PLR, and LMR in patients with breast cancer compared to healthy controls, and to explore their potential utility as inflammatory biomarkers for early detection of the disease in our local population. Materials and Methods Study Design and Participants This case-control study was conducted at PHO Clinical hospital Dr. Trifun Panovski Bitola, North Macedonia, between January to June 2025 year. In this study we include a total of 62 female patients with breast cancer. The control group consisted of age-matched healthy females with no history of malignancy or inflammatory disease. Inclusion and Exclusion Criteria Patients with active infections, autoimmune diseases, hematological disorders, or on immunosuppressive therapy were excluded to avoid confounding effects on inflammatory markers. Controls were screened to exclude any acute or chronic illnesses. Data Collection and Laboratory Analysis Peripheral blood samples were collected from all participants under standardized conditions. Complete blood counts (CBC) were performed using an automated hematology analyzer Sysmeh XN 550. The following parameters were recorded: neutrophil count, lymphocyte count, monocyte count, and platelet count. The inflammatory ratios were calculated as follows: Neutrophil-to-lymphocyte ratio (NLR) = neutrophil count / lymphocyte count Platelet-to-lymphocyte ratio (PLR) = platelet count / lymphocyte count Lymphocyte-to-monocyte ratio (LMR) = lymphocyte count / monocyte count Statistical Analysis Statistical analysis were performed using SPSS version 13. For continuous variables, data were summarized either as mean ± standard deviation (SD) or as median with interquartile range (IQR), depending on the shape of the data distribution. The Shapiro-Wilk test was used to determine whether the data followed a normal distribution. Group comparisons were carried out using either the Student’s t-test for normally distributed variables or the Mann-Whitney U test when the normality assumption was not met. The diagnostic performance of the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR) was evaluated through receiver operating characteristic (ROC) curve analysis. A p-value below 0.05 was considered statistically significant throughout the study. Results A total of 62 participants were included in the study, evenly divided into two groups: 31 patients with breast cancer (Group A) and 31 healthy women as controls (Group B). The median age was 61 years in both groups, with the youngest participant aged 44 and the oldest 83 years Tab.1. Table 1. Display and comparison of the evaluated parameters of our study sample. Mean values Group A Group B P value (Breast cancer) n=31 (Control group) N=31 Age (years) 61(44–77) 61 (44–83) 0.391 PLR (%) 136.96 (49.34–308.7) 118.8 (66.03–215.38) 0.087 NLR (%) 1.82 (0.34–6.38) 1.66 (0.89–3.18) 0.477 LMR(%) 4.35 (0.68–11) 4.88 (2.38–7.37) 0.141 PLR – platelets count/lymphocytes ratio; NLR – neutrophils/lymphocytes ratio; LMR - lymphocytes/monocytes ratio When comparing inflammatory markers, no statistically significant differences were observed between breast cancer patients and healthy controls in platelet-to-lymphocyte ratio (PLR) Fig.1, neutrophil-to-lymphocyte ratio (NLR) Fig.2, and lymphocyte-to-monocyte ratio (LMR) Fig.3. Table 2. Hematological parameters of the study populations Mean values Group A Breast cancer patients n=31 Group B Control group n=31 P-value RBC (x10 12 /l) 4.29±0.48 4.76±0.41 0.317 Hgb(g/l) 129.68±15.25 140.65±11.77 0.001 Hct(L/L) 0.39±0.04 0.42±0.03 0.447 MCV(fl) 91.39±3.8 88.65±4.75 0.428 MCH(pg) 29.5±4.24 29.61±1.83 0.444 WBC (×10 9 /l) 6.5±1.93 6.87±1.51 0.467 PC (×10 9 /l) 265.45±79.67 268.65±53.13 0.426 Neutrophils(×10 9 /l) 3.5±1.28 3.77±1.07 0.146 Lymphocytes(×10 9 /l) 2.24±0.89 2.42±0.74 0.041 Monocytes(×10 9 /l) 0.58±0.2 0.51±0.12 0.223 Éosinophiles(×10 9 /l) 0.15±0.12 0.15±0.11 0.377 RBC - red blood cells, Hgb - hemoglobin, Hct - hematoctit, MCV - mean corpuscular volume, MCH - mean corpuscular haemoglobin, PC – platelets count; WBC – white blood cell count However, breast cancer patients demonstrated a statistically significant increase in lymphocyte count compared to the control group (p = 0.04). Additionally, hemoglobin levels were significantly lower in breast cancer patients relative to healthy controls (p = 0.001). No statistically significant differences were found between the two groups for red blood cell count, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), platelet count, white blood cell count, neutrophils, monocytes, and eosinophils Tab.2. Discussion Previous studies have reported conflicting results regarding the differences in neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR) among patients with breast cancer (BC), those with benign breast conditions, and healthy individuals (18–22). This inconsistency has created uncertainty about the diagnostic value of these inflammatory markers for BC screening and early detection. The link between inflammation and cancer was first noted by Rudolf Virchow, who identified leukocytes within tumors and hypothesized that inflammation promotes cellular proliferation (23). Since then, inflammation has been established as a hallmark of cancer and a key biological process involved in tumor initiation, progression, and metastasis (24, 25). Neutrophils, in particular, play a complex dual role by both inhibiting and facilitating tumor progression, which has drawn significant research interest concerning their role within the tumor microenvironment (26). An imbalance in NLR has been associated with enhanced tumor progression and metastasis, with NLR serving as an accessible marker of cancer-related inflammation due to neutrophil stimulation. Several recent studies have linked elevated NLR to poorer prognosis in various cancers and have underscored its importance in metastasis and recurrence (27, 28). These findings highlight the critical need for further investigation into the mechanistic role of NLR in cancer development. The interplay between inflammation, coagulation, and tumor progression remains an active area of research, as these processes form vicious cycles that promote tumor growth. Although the exact pathophysiological mechanisms are not fully understood, there is increasing clinical interest in integrating these parameters as novel biomarkers in oncology (29, 30). NLR and PLR are emerging as convenient inflammatory biomarkers in clinical practice (31). While the prognostic significance of NLR in solid tumors is well supported by robust data (32–34), the prognostic value of PLR remains controversial, with studies reporting mixed results regarding its accuracy (35–38). Platelet activation is known to contribute to tumor growth by promoting neoangiogenesis, degrading the extracellular matrix, and releasing adhesion molecules and growth factors essential for metastasis (39–41). In our study, we did not observe statistically significant differences in PLR, NLR, or LMR between breast cancer patients and healthy controls, consistent with findings by Yang et al. (42). However, we found significantly elevated lymphocyte counts in breast cancer patients, aligning with the study by Obeagu (43). This elevation may reflect an immune response against tumor cells, suggesting that lymphocyte count alone could have potential clinical relevance in BC. Overall, our results contribute to the growing body of evidence indicating that while inflammatory markers like NLR and PLR may have prognostic value in some cancers, their diagnostic role in breast cancer requires further exploration. Larger, prospective studies are needed to clarify these associations and to better define the clinical utility of these hematologic parameters in breast cancer management. Conclusion In this small study comparing breast cancer patients and healthy controls, we observed a significant increase in lymphocyte counts among breast cancer patients. However, no statistically significant differences were found in the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), or lymphocyte-to-monocyte ratio (LMR) between the two groups. These findings suggest that while lymphocyte levels may have potential relevance in breast cancer, the utility of NLR, PLR, and LMR as diagnostic markers requires further investigation in larger studies. Declarations Ethics Approval Statement This study was reviewed and approved by the Ethics Committee at PHO Clinical Hospital Dr. Trifun Panovski, Bitola, Republic of Macedonia, under approval number 12212/02/2025. All research procedures involving human participants were conducted in accordance with institutional ethical standards and the principles of the Declaration of Helsinki (1964) and its subsequent amendments. Consent Consent was obtained for all participants Ethical Approval and Consent to Participate This study was approved by the Ethics Committee of PHO Clinical Hospital “Dr. Trifun Panovski” Bitola, North Macedonia. Written informed consent to participate was obtained from all individuals included in the study. Consent for Publication Not applicable. This manuscript does not contain any individual person’s data in any form (including images, videos, or identifiable personal details). Funding This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors. Data Availability The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request. 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Prognostic value of systemic immune-inflammation index in cancer: A meta-analysis. J Cancer. 2018;9:3295–302. Obeagu EI, Obeagu GU. Lymphocyte infiltration in breast cancer: A promising prognostic indicator. Medicine (Baltimore). 2024;103(49):e40845. 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-7092201","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":484962582,"identity":"8a8b245f-1c45-4252-afcf-a1d455d98945","order_by":0,"name":"Biljana Ilkovska","email":"data:image/png;base64,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","orcid":"","institution":"University St. Kliment Ohridski Bitola, Higher Medical School Bitola, Department of Laboratory Diagnostics, PHO Clinical Hospital Dr. Trifun Panovski, Bitola, R. 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Despite advances in diagnosis and treatment, a significant proportion of patients still experience recurrence or distant metastases within five years of initial diagnosis [2,3].\u003c/p\u003e\n\u003cp\u003eSystemic inflammation has been increasingly recognized as a critical component in the pathogenesis and progression of cancer, including BC [4\u0026ndash;6]. Tumor cells can provoke systemic inflammatory responses, leading to measurable changes in hematologic parameters such as neutrophils, lymphocytes, platelets, and monocytes [7]. These inflammatory changes may be reflected in specific ratios derived from complete blood counts\u0026mdash;namely the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR)\u0026mdash;which have been investigated as potential diagnostic and prognostic biomarkers in several solid tumors, including breast cancer [8\u0026ndash;10].\u003c/p\u003e\n\u003cp\u003eThese markers are inexpensive, routinely available, and non-invasive, making them especially valuable in low- and middle-income countries where access to advanced diagnostic tools may be limited [11,12]. While several studies have supported the utility of NLR, PLR, and LMR in the prognosis of BC, findings have been inconsistent, particularly regarding their diagnostic value [13\u0026ndash;15].\u003c/p\u003e\n\u003cp\u003eIn North Macedonia, data on the association between these hematological indices and breast cancer are scarce. Therefore, the aim of this study was to evaluate the levels of NLR, PLR, and LMR in patients with breast cancer compared to healthy controls, and to explore their potential utility as inflammatory biomarkers for early detection of the disease in our local population.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cstrong\u003eStudy Design and Participants\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis case-control study was conducted at PHO Clinical hospital Dr. Trifun Panovski Bitola, North Macedonia, between January to June 2025 year. In this study we include a total of 62 female patients with breast cancer. The control group consisted of age-matched healthy females with no history of malignancy or inflammatory disease.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInclusion and Exclusion Criteria\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePatients with active infections, autoimmune diseases, hematological disorders, or on immunosuppressive therapy were excluded to avoid confounding effects on inflammatory markers. Controls were screened to exclude any acute or chronic illnesses.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Collection and Laboratory Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePeripheral blood samples were collected from all participants under standardized conditions. Complete blood counts (CBC) were performed using an automated hematology analyzer Sysmeh XN 550. The following parameters were recorded: neutrophil count, lymphocyte count, monocyte count, and platelet count.\u003c/p\u003e\n\u003cp\u003eThe inflammatory ratios were calculated as follows:\u003c/p\u003e\n\u003cp\u003eNeutrophil-to-lymphocyte ratio (NLR) = neutrophil count / lymphocyte count\u003c/p\u003e\n\u003cp\u003ePlatelet-to-lymphocyte ratio (PLR) = platelet count / lymphocyte count\u003c/p\u003e\n\u003cp\u003eLymphocyte-to-monocyte ratio (LMR) = lymphocyte count / monocyte count\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStatistical analysis were performed using SPSS version 13. For continuous variables, data were summarized either as mean \u0026plusmn; standard deviation (SD) or as median with interquartile range (IQR), depending on the shape of the data distribution. The Shapiro-Wilk test was used to determine whether the data followed a normal distribution. Group comparisons were carried out using either the Student\u0026rsquo;s t-test for normally distributed variables or the Mann-Whitney U test when the normality assumption was not met. The diagnostic performance of the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR) was evaluated through receiver operating characteristic (ROC) curve analysis. A p-value below 0.05 was considered statistically significant throughout the study.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eA total of 62 participants were included in the study, evenly divided into two groups: 31 patients with breast cancer (Group A) and 31 healthy women as controls (Group B). The median age was 61 years in both groups, with the youngest participant aged 44 and the oldest 83 years Tab.1.\u003c/p\u003e\n\u003ch3\u003eTable 1. Display and comparison of the evaluated parameters of our study sample.\u003c/h3\u003e\n\u003ctable border=\"0\" cellpadding=\"0\" width=\"500\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean values\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroup A\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroup B\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u003cstrong\u003e\u0026nbsp;value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e(Breast cancer)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003en=31\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e(Control group)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eN=31\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAge (years)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e61(44\u0026ndash;77)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e61 (44\u0026ndash;83)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.391\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePLR (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e136.96 (49.34\u0026ndash;308.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e118.8 (66.03\u0026ndash;215.38)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.087\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNLR (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1.82 (0.34\u0026ndash;6.38)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1.66 (0.89\u0026ndash;3.18)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.477\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eLMR(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e4.35 (0.68\u0026ndash;11)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e4.88 (2.38\u0026ndash;7.37)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.141\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003ePLR \u0026ndash; platelets count/lymphocytes ratio; NLR \u0026ndash; neutrophils/lymphocytes ratio; LMR - lymphocytes/monocytes ratio\u003c/p\u003e\n\u003cp\u003eWhen comparing inflammatory markers, no statistically significant differences were observed between breast cancer patients and healthy controls in platelet-to-lymphocyte ratio (PLR) Fig.1, neutrophil-to-lymphocyte ratio (NLR) Fig.2, and lymphocyte-to-monocyte ratio (LMR) Fig.3.\u003c/p\u003e\n\u003cp\u003eTable 2. Hematological parameters of the study populations\u003c/p\u003e\n\u003ctable border=\"0\" cellpadding=\"0\" width=\"576\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean values\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroup A\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eBreast cancer patients\u003cbr\u003e\u0026nbsp;n=31\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroup B\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eControl group\u003cbr\u003e\u0026nbsp;n=31\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eRBC (x10\u003csup\u003e12\u003c/sup\u003e/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e4.29\u0026plusmn;0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e4.76\u0026plusmn;0.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.317\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eHgb(g/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e129.68\u0026plusmn;15.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e140.65\u0026plusmn;11.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eHct(L/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.39\u0026plusmn;0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.42\u0026plusmn;0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.447\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eMCV(fl)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e91.39\u0026plusmn;3.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e88.65\u0026plusmn;4.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.428\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eMCH(pg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e29.5\u0026plusmn;4.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e29.61\u0026plusmn;1.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.444\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eWBC (\u0026times;10\u003csup\u003e9\u003c/sup\u003e/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6.5\u0026plusmn;1.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6.87\u0026plusmn;1.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.467\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePC (\u0026times;10\u003csup\u003e9\u003c/sup\u003e/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e265.45\u0026plusmn;79.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e268.65\u0026plusmn;53.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.426\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNeutrophils(\u0026times;10\u003csup\u003e9\u003c/sup\u003e/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e3.5\u0026plusmn;1.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e3.77\u0026plusmn;1.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.146\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eLymphocytes(\u0026times;10\u003csup\u003e9\u003c/sup\u003e/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e2.24\u0026plusmn;0.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e2.42\u0026plusmn;0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.041\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eMonocytes(\u0026times;10\u003csup\u003e9\u003c/sup\u003e/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.58\u0026plusmn;0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.51\u0026plusmn;0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.223\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026Eacute;osinophiles(\u0026times;10\u003csup\u003e9\u003c/sup\u003e/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.15\u0026plusmn;0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.15\u0026plusmn;0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.377\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eRBC - red blood cells, Hgb - hemoglobin, Hct - hematoctit, MCV - mean corpuscular volume, MCH - mean corpuscular haemoglobin, PC \u0026ndash; platelets count; \u0026nbsp;WBC \u0026ndash; white blood cell count\u003c/p\u003e\n\u003cp\u003eHowever, breast cancer patients demonstrated a statistically significant increase in lymphocyte count compared to the control group (p = 0.04). Additionally, hemoglobin levels were significantly lower in breast cancer patients relative to healthy controls (p = 0.001).\u003c/p\u003e\n\u003cp\u003eNo statistically significant differences were found between the two groups for red blood cell count, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), platelet count, white blood cell count, neutrophils, monocytes, and eosinophils Tab.2.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003ePrevious studies have reported conflicting results regarding the differences in neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR) among patients with breast cancer (BC), those with benign breast conditions, and healthy individuals (18\u0026ndash;22). This inconsistency has created uncertainty about the diagnostic value of these inflammatory markers for BC screening and early detection.\u003c/p\u003e\n\u003cp\u003eThe link between inflammation and cancer was first noted by Rudolf Virchow, who identified leukocytes within tumors and hypothesized that inflammation promotes cellular proliferation (23). Since then, inflammation has been established as a hallmark of cancer and a key biological process involved in tumor initiation, progression, and metastasis (24, 25). Neutrophils, in particular, play a complex dual role by both inhibiting and facilitating tumor progression, which has drawn significant research interest concerning their role within the tumor microenvironment (26).\u003c/p\u003e\n\u003cp\u003eAn imbalance in NLR has been associated with enhanced tumor progression and metastasis, with NLR serving as an accessible marker of cancer-related inflammation due to neutrophil stimulation. Several recent studies have linked elevated NLR to poorer prognosis in various cancers and have underscored its importance in metastasis and recurrence (27, 28). These findings highlight the critical need for further investigation into the mechanistic role of NLR in cancer development.\u003c/p\u003e\n\u003cp\u003eThe interplay between inflammation, coagulation, and tumor progression remains an active area of research, as these processes form vicious cycles that promote tumor growth. Although the exact pathophysiological mechanisms are not fully understood, there is increasing clinical interest in integrating these parameters as novel biomarkers in oncology (29, 30).\u003c/p\u003e\n\u003cp\u003eNLR and PLR are emerging as convenient inflammatory biomarkers in clinical practice (31). While the prognostic significance of NLR in solid tumors is well supported by robust data (32\u0026ndash;34), the prognostic value of PLR remains controversial, with studies reporting mixed results regarding its accuracy (35\u0026ndash;38). Platelet activation is known to contribute to tumor growth by promoting neoangiogenesis, degrading the extracellular matrix, and releasing adhesion molecules and growth factors essential for metastasis (39\u0026ndash;41).\u003c/p\u003e\n\u003cp\u003eIn our study, we did not observe statistically significant differences in PLR, NLR, or LMR between breast cancer patients and healthy controls, consistent with findings by Yang et al. (42). However, we found significantly elevated lymphocyte counts in breast cancer patients, aligning with the study by Obeagu (43). This elevation may reflect an immune response against tumor cells, suggesting that lymphocyte count alone could have potential clinical relevance in BC.\u003c/p\u003e\n\u003cp\u003eOverall, our results contribute to the growing body of evidence indicating that while inflammatory markers like NLR and PLR may have prognostic value in some cancers, their diagnostic role in breast cancer requires further exploration. Larger, prospective studies are needed to clarify these associations and to better define the clinical utility of these hematologic parameters in breast cancer management.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn this small study comparing breast cancer patients and healthy controls, we observed a significant increase in lymphocyte counts among breast cancer patients. However, no statistically significant differences were found in the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), or lymphocyte-to-monocyte ratio (LMR) between the two groups. These findings suggest that while lymphocyte levels may have potential relevance in breast cancer, the utility of NLR, PLR, and LMR as diagnostic markers requires further investigation in larger studies.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eEthics Approval Statement This study was reviewed and approved by the Ethics Committee at PHO Clinical Hospital Dr. Trifun Panovski, Bitola, Republic of Macedonia, under approval number 12212/02/2025. All research procedures involving human participants were conducted in accordance with institutional ethical standards and the principles of the Declaration of Helsinki (1964) and its subsequent amendments.\u003c/p\u003e\n\u003cp\u003eConsent Consent was obtained for all participants\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eEthical Approval and Consent to Participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was approved by the Ethics Committee of PHO Clinical Hospital \u0026ldquo;Dr. Trifun Panovski\u0026rdquo; Bitola, North Macedonia. Written informed consent to participate was obtained from all individuals included in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for Publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. This manuscript does not contain any individual person\u0026rsquo;s data in any form (including images, videos, or identifiable personal details).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eSung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. 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Medicine (Baltimore). 2024;103(49):e40845.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"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, NLR, PLR, LMR, inflammation, biomarkers, hematology, early detection","lastPublishedDoi":"10.21203/rs.3.rs-7092201/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7092201/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e\u003cbr\u003e\nBreast cancer (BC) is one of the most common malignancies among women worldwide, with approximately 2.3 million new cases and 685,000 deaths reported in 2020. Inflammation-related hematologic parameters—such as the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR)—have been proposed as potential biomarkers for cancer progression. However, data on their association with breast cancer in North Macedonia are limited.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAim:\u003c/strong\u003e\u003cbr\u003e\nThis study aimed to evaluate NLR, PLR, and LMR in patients with breast cancer compared to healthy controls, to explore their potential as inflammatory biomarkers for early detection.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e\u003cbr\u003e\nA case-control study was conducted in 2025 at the Department of Medical Biochemistry, Clinical Hospital \"Dr. Trifun Panovski\" in Bitola. Sixty-two women were enrolled -31 with histologically confirmed breast cancer and 31 age-matched healthy controls. Hematological parameters were measured using a Sysmex XP-300 hematology analyzer. Statistical analyses were performed using SPSS v13.0.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e\u003cbr\u003e\nThe mean lymphocyte count was significantly higher in breast cancer patients compared to controls (p = 0.041). No statistically significant differences were found in PLR (p = 0.087), NLR (p = 0.477), or LMR (p = 0.141) between groups. Hemoglobin levels were significantly lower in the breast cancer group (p = 0.001).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e\u003cbr\u003e\nWhile lymphocyte levels and hemoglobin were significantly altered in breast cancer patients, our results did not support NLR, PLR, or LMR as standalone markers for early breast cancer detection in this population. Further large-scale studies are needed to validate these findings.\u003c/p\u003e","manuscriptTitle":"Comparative Analysis of Inflammatory Biomarkers and Lymphocyte Levels in Breast Cancer Patients and Healthy Controls","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-18 12:33:51","doi":"10.21203/rs.3.rs-7092201/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":"26935462-462d-46db-ba8b-0675e5af3a2c","owner":[],"postedDate":"July 18th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-07-18T12:33:53+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-18 12:33:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7092201","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7092201","identity":"rs-7092201","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}