Chemerin Promotes Oral Squamous Cell Carcinoma Progression via NLRP3-Mediated Pyroptosis and the GSDMD-N Pathway.

Conceptualisation: Feng, N. Wang, Zhang. Methodology: Feng, N. Wang, Zhang. Validation: Feng, Gao, Guo, Li, Lin, Ni, N. Wang, Zhang. Investigation: Feng, N. Wang, Zhang. Formal analysis: Gao, Guo, Li, Lin, Ni. Funding acquisition: Tan, C. Wang, N. Wang. Writing – original draft: Feng, N. Wang, Zhang. The first draft was prepared by Feng, N. Wang and Zhang. All authors have critically reviewed, revised and approved the final version of the manuscript.

The Cancer Genome Atlas (TCGA) database was searched, and the expression of chemerin and GSDMD mRNA in head and neck squamous cell carcinoma (HNSC) and normal squamous epithelial tissues was analysed. The results indicated that chemerin ( P .05) expression in HNSC tissues was higher in HNSC tissues compared to those in normal squamous epithelial tissues ( Figure 1A ). Fig. 1 Expression of pyroptosis-related proteins in OSCC tissues: (a, b, c, e, f and g) OSCC tissue and (d and h) normal mucosal tissue. A, Expression of chemerin and GSDMD in HNSC and normal squamous epithelial tissues in the TCGA database. B, Expression of GSDMD in OSCC tissues and normal mucosal tissues. C, Expression of IL-1β in OSCC tissues and normal mucosal tissues. Fig 1 dummy alt text Expression of pyroptosis-related proteins in OSCC tissues: (a, b, c, e, f and g) OSCC tissue and (d and h) normal mucosal tissue. A, Expression of chemerin and GSDMD in HNSC and normal squamous epithelial tissues in the TCGA database. B, Expression of GSDMD in OSCC tissues and normal mucosal tissues. C, Expression of IL-1β in OSCC tissues and normal mucosal tissues. The IHC staining results indicated that the pyroptosis-related proteins GSDMD and IL-1β were predominantly localised in the cytoplasm ( Fig. 1 , Fig. 1 ). The analysis results indicated that in OSCC samples, 77 cases (53.1%) had strong expression of GSDMD, and 68 cases (46.9%) had weak expression. Most of the 16 cases of normal mucosal tissue samples (12/16, 75%) had weak or even negative expression; only 25% (4/16) showed moderate expression ( Figure 1B ). In addition, IL-1β had strong expression in 80 cases (55.17%) and weak expression in 65 cases (44.83%) in OSCC samples; weak and negative expressions (13/16, 81.25%) were mainly found in normal mucosal tissues; a small number (3/16, 18.75%) of tissues achieved moderate expression ( Figure 1C ). The results indicated that high GSDMD expression was associated with lower differentiation, lymph node metastasis, a later stage and tumour recurrence. Furthermore, no correlation was observed between GSDMD expression and factors such as gender, age or tumour diameter. ( Table 1 ). Table 1 Relationship between GSDMD expression and clinicopathological parameters in OSCC. Table 1 dummy alt text Variables n GSDMD χ² P High(%) Low(%) Gender Male 79 45(57.0) 34(43.0) 0.085 .770 Female 66 36(54.5) 30(45.5) Age ≥60 51 28(54.9) 23(45.1) 0.029 .864 <60 94 53(56.4) 41(43.6) Differentiation G1 80 35(43.8) 45(56.2) 10.618 .001 ⁎⁎ G2, G3 65 46(70.8) 19(29.2) Lymph node metastasis Yes 66 46(69.7) 20(30.3) 9.339 .002 ⁎⁎ No 79 35(44.3) 44(55.7) TNM stage Ⅰ, Ⅱ 71 31(43.7) 40(56.3) 8.398 .004 ⁎⁎ Ⅲ, Ⅳ 74 50(67.6) 24(32.4) Tumour size <4 103 56(54.4) 47(45.6) 0.322 .571 ≥4 42 25(59.5) 17(40.5) Tumour recurrence Yes 109 67(61.5) 42(38.5) 5.596 .018* No 36 14(38.9) 22(61.1) * P <.05; ⁎⁎ P <.01. Relationship between GSDMD expression and clinicopathological parameters in OSCC. * P <.05; ⁎⁎ P <.01. The overall survival (OS) and disease-free survival (DFS) times of 145 patients were analysed. The Kaplan–Meier analysis results indicated that the average OS and DFS time of patients with low GSDMD expression level were 91.10 and 88.46 months, respectively, whereas those with high GSDMD expression were 52.79 and 49.56 months, respectively. This difference is significant ( P < .001). In addition, among patients with moderately and poorly differentiated OSCC, the OS and DFS time of the group with higher GSDMD expression were 39.75 and 34.65 months, respectively, whereas those of the group with lower GSDMD expression were 90.26 and 85.10 months, respectively. This discrepancy is noteworthy ( P = .003; P = .022). Among patients with lymph node metastasis, the OS (40.18 months) and DFS (36.35 months) times of patients with high GSDMD expression were shorter than those observed in patients with low GSDMD expression (47.40 and 45.41 months; P = .017, P = .048; Figure 2 ). Fig. 2 Relationship between GSDMD expression and patient postoperative survival time. A, In the group with high GSDMD expression level in OSCC tissues, the OS and DFS times were shorter. B, Among moderately and poorly differentiated patients, the group with high GSDMD expression level has a shorter OS and DFS time. C, Among patients with lymph node metastasis, the group with higher GSDMD expression level has a shorter OS and DFS time. Fig 2 dummy alt text Relationship between GSDMD expression and patient postoperative survival time. A, In the group with high GSDMD expression level in OSCC tissues, the OS and DFS times were shorter. B, Among moderately and poorly differentiated patients, the group with high GSDMD expression level has a shorter OS and DFS time. C, Among patients with lymph node metastasis, the group with higher GSDMD expression level has a shorter OS and DFS time. Single-factor and multifactor analyses were used to determine the independent prognostic factors affecting patient prognosis. The results indicated that GSDMD expression, differentiation, lymph node metastasis, and TNM stage were the factors affecting the OS time of patients with OSCC ( Table 2 ). The factors affecting patients’ DFS time included GSDMD expression, differentiation, lymph node metastasis, TNM stage and tumour recurrence ( Table 3 ). The statistically significant single factors were subjected to multifactor analysis, and the results indicated that GSDMD expression is an indicator affecting the prognosis of OS time for patients with OSCC ( Table 2 ). GSDMD expression and tumour recurrence are independent prognostic factors affecting the DFS time of patients with OSCC ( Table 3 ). Table 2 Univariate and multivariate Cox regression model analysis of factors affecting the overall survival time of OSCC patients. Table 2 dummy alt text Variables Univariate analysis Multivariate analysis HR (95%CI) P HR (95%CI) P Gender 1.334 (0.814-2.186) .254 - - Age 1.026 (0.614-1.712) .923 - - Differentiation 0.465 (0.285-0.759) .002 - - Lymph node metastasis 2.644 (1.608-4.349) <.001 - - TNM stage 0.327 (0.195-0.550) <.001 - - Tumour size 0.918 (0.534-1.579) .758 - - Tumour recurrence 0.908 (0.516-1.600) .739 - - GSDMD expression 3.121 (1.793-5.433) <.001 2.306 (1.288-4.128) .005 Table 3 Univariate and multivariate Cox regression model analysis of factors affecting disease-free survival time of OSCC patients. Table 3 dummy alt text Variables Univariate analysis Multivariate analysis HR (95%CI) P HR (95%CI) P Gender 1.263 (0.771-2.072) .354 - - Age 1.245 (0.745-2.081) .402 - - Differentiation 0.440 (0.269-0.720) .001 - - Lymph node metastasis 2.496 (1.517-4.107) <.001 - - TNM stage 0.346 (0.205-0.583) <.001 - - Tumour size 0.866 (0.504-1.489) .603 - - Tumour recurrence 0.556 (0.314-0.987) .045 0.368 (0.200-0.677) .001 GSDMD expression 2.946 (1.691-5.133) <.001 2.616 (1.399-4.892) .003 Univariate and multivariate Cox regression model analysis of factors affecting the overall survival time of OSCC patients. Univariate and multivariate Cox regression model analysis of factors affecting disease-free survival time of OSCC patients. The immunohistochemical result indicated that tumour areas with strong chemerin expression also had relatively strong GSDMD expression, and vice versa. GSDMD expression was also reduced in areas with lower chemerin expression levels ( Figure 3A ). In addition, the result of immunofluorescence double staining revealed that chemerin and GSDMD protein were coexpressed and localised in OSCC tissues ( Figure 3B ). Spearmen’s correlation analysis revealed a positive correlation between the expression of chemerin and GSDMD (r = 0.229, P = .006; Table 4 ). Fig. 3 Detection of chemerin and GSDMD expression. A, The results indicate that GSDMD was highly expressed in areas with high chemerin expression level; when chemerin expression was low, GSDMD was also low. B, The results of immunofluorescence double staining indicate that chemerin (488 nm) and GSDMD (650 nm) can be jointly expressed and localised in OSCC (red for chemerin and green for GSDMD). Fig 3 dummy alt text Table 4 Correlation between Chemerin expression and GSDMD expression. Table 4 dummy alt text GSDMD expression Chemerin expression Spearman’s rho coefficient test High Low r P High 68 13 0.229 .006 ⁎⁎ Low 41 23 ⁎⁎ P <.01. Detection of chemerin and GSDMD expression. A, The results indicate that GSDMD was highly expressed in areas with high chemerin expression level; when chemerin expression was low, GSDMD was also low. B, The results of immunofluorescence double staining indicate that chemerin (488 nm) and GSDMD (650 nm) can be jointly expressed and localised in OSCC (red for chemerin and green for GSDMD). Correlation between Chemerin expression and GSDMD expression. ⁎⁎ P <.01. The basic expressions of chemerin in SCC9, SCC15 and Cal27 cells were assessed using Western blot and qPCR. The findings indicated that the expression of chemerin was significantly elevated in SCC15 cells, whereas lower levels were observed in both SCC9 and Cal27 cells ( Figure 4A ). Although chemerin expression in SCC9 was a little lower than that in Cal-27, SCC9 cells showed poor viability following lentiviral transfection. We had adjusted the MOI value for transfection several times, but the state of SCC9 was still not satisfactory. Therefore, the knockdown lentivirus was transfected into SCC15 cells to interfere with the expression of chemerin, and lentivirus overexpressing chemerin was used to transfect Cal27 cells with low expression. The transfection efficiency of chemerin was detected by Western blot and qPCR ( Figure 4A - C ). Fig. 4 Lentiviral transfection efficiency in OSCC and the effect of chemerin expression on the pyroptosis of cells. GAPDH was used as a control. A, a, Basal expression of chemerin in 3 cell lines (SCC9, SCC15 and Cal27). A, b, qPCR was used to detect the transfection efficiency of SCC15 and Cal27 cell. A, c, The transfection efficiency of the SCC15 and Cal27 cell lines was detected by Western blot. B, Chemerin overexpression (Cal27-Chemerin) can increase the expression level of GSDMD, GSDMD-N and IL-1β; chemerin knockdown (SCC15-Chemerin-shRNA) can decrease the expression level of GSDMD, GSDMD-N and IL-1β. C, The expression of IL-1β in the cell supernatant was detected by ELISA. Fig 4 dummy alt text Lentiviral transfection efficiency in OSCC and the effect of chemerin expression on the pyroptosis of cells. GAPDH was used as a control. A, a, Basal expression of chemerin in 3 cell lines (SCC9, SCC15 and Cal27). A, b, qPCR was used to detect the transfection efficiency of SCC15 and Cal27 cell. A, c, The transfection efficiency of the SCC15 and Cal27 cell lines was detected by Western blot. B, Chemerin overexpression (Cal27-Chemerin) can increase the expression level of GSDMD, GSDMD-N and IL-1β; chemerin knockdown (SCC15-Chemerin-shRNA) can decrease the expression level of GSDMD, GSDMD-N and IL-1β. C, The expression of IL-1β in the cell supernatant was detected by ELISA. In exploring the effect of chemerin expression on pyroptosis, the expression of pyroptosis-related proteins was detected by Western blot. The results demonstrated a significant increase in the expression levels of GSDMD-N, GSDMD and IL-β within the Cal27-Chemerin group. Conversely, a notable decrease was observed in the SCC15-Chemerin-shRNA group ( Figure 4B ). In addition, the ELISA results indicated that after overexpression of chemerin in Cal27 cells, there was a notable increase in the levels of IL-1β present in the cell supernatant, and vice versa. The samples were divided into 3 groups: group 1, strong chemerin expression and strong GSDMD expression; group 2, strong chemerin expression, weak GSDMD expression or weak chemerin expression, and strong GSDMD expression; group 3, weak chemerin expression and weak GSDMD expression. In OSCC, strong chemerin expression and strong GSDMD expression were related to lower differentiation, lymph node metastasis, later stage and tumour recurrence ( P < .05, Table 5 ). Table 5 Relationship between Chemerin and GSDMD expression and clinicopathological parameters in OSCC. Table 5 dummy alt text Variables n Group 1 Group 2 Group 3 χ² P Gender Male 79 41(51.9) 24(30.4) 14 (17.7) 3.499 .174 Female 66 27(40.9) 30(45.5) 9 (13.6) Age ≥60 51 25(49.0) 21(41.2) 5 (9.8) 2.223 .329 <60 94 43 (45.7) 33(37.2) 18 (17.0) Differentiation G1 80 28 (35.0) 36(45.0) 16 (20.0) 10.197 .006 ⁎⁎ G2, G3 65 40 (61.5) 18(27.7) 7 (10.8) Lymph node metastasis Yes 66 40(60.6) 19(28.8) 7 (10.6) 9.289 .01* No 79 28(35.4) 35(44.3) 16 (20.3) TNM stage Ⅰ, Ⅱ 71 26(36.6) 29(40.8) 16 (22.5) 7.524 .023* Ⅲ, Ⅳ 74 42(56.8) 25(33.8) 7 (9.5) Tumour size <4 103 44(42.7) 43(41.7) 16 (15.5) 3.287 0.193 ≥4 42 24(57.1) 11(26.2) 7 (16.7) Tumour recurrence Yes 109 57(52.3) 30(27.5) 22 (20.2) 19.030 .000 ⁎⁎⁎ No 36 11(30.6) 24(66.7) 1 (2.8) * P <.05; ⁎⁎ P <.01; ⁎⁎⁎ P <.001. Relationship between Chemerin and GSDMD expression and clinicopathological parameters in OSCC. * P <.05; ⁎⁎ P <.01; ⁎⁎⁎ P <.001. Kaplan–Meier analysis indicated that the average OS time (53.98 months) of patients with strong chemerin expression and strong GSDMD expression was significantly shorter than the average OS time of group 2 (80.23 months) and group 3 (88.88 months). This difference was statistically significant ( P = .003). The average DFS time of patients in group 1 was 50.31 months; the average DFS time of patients in group 2 was 76.55 months; and the average DFS time of patients in group 3 was 89.13 months. The DFS time for group 1 was significantly higher than the other 2 groups ( P = .005, Figure 5 ). Fig. 5 Relationship between the expression of chemerin and GSDMD on the OS and DFS time of patients with OSCC. The group with a high expression level of chemerin and GSDMD had shorter OS and DFS time than the other 2 groups. Note: group 1: strong chemerin expression and strong GSDMD expression; group 2: strong chemerin expression, weak GSDMD expression or weak chemerin expression and strong GSDMD expression; group 3: weak chemerin expression and weak GSDMD expression. Fig 5 dummy alt text Relationship between the expression of chemerin and GSDMD on the OS and DFS time of patients with OSCC. The group with a high expression level of chemerin and GSDMD had shorter OS and DFS time than the other 2 groups. Note: group 1: strong chemerin expression and strong GSDMD expression; group 2: strong chemerin expression, weak GSDMD expression or weak chemerin expression and strong GSDMD expression; group 3: weak chemerin expression and weak GSDMD expression. IHC staining revealed that the expression of NLRP3 was also relatively high in areas with high expression of chemerin, and vice versa, and the expression of NRLP3 was weak in areas with low expression of chemerin ( Figure 6A ). Meanwhile, the results of immunofluorescence double staining indicated that chemerin and NLRP3 protein were coexpressed and localised in OSCC tissues ( Figure 6B ). Spearmen correlation analysis showed that the expression of chemerin was positively correlated with that of NLRP3 ( Table 6 ). Cytological experiments showed that the expression of NLRP3 was significantly increased in Cal27 with chemerin overexpression but decreased in SCC15 with chemerin knockdown. When the NLRP3 inhibitor MCC950 was added to each group, the expression of pyroptosis-related proteins (NLRP3, Caspase-1, GSDMD, GSDMD-N, IL-1β and caspase-1) decreased significantly ( Figure 6C ). After adding MCC950, the expression of pyroptosis-related proteins was relatively reduced in all groups ( Figure 6D ). Fig. 6 Expression of chemerin and NLRP3 in OSCC. A, The results indicated that NLRP3 was highly expressed in the high-expression region of chemerin. B, Immunofluorescence double staining showed that chemerin (488 nm) and NLRP3 (650 nm) could be coexpressed and localised in OSCC cells (red for chemerin and green for NLRP3). C, Chemerin overexpression led to the increased expression of pyroptosis-related proteins, and vice versa. When the NLRP3 inhibitor (MCC950) was added to each group, the expression level of pyroptosis-related proteins was decreased. GAPDH was used as a control. Fig 6 dummy alt text Table 6 Correlation analysis of chemerin expression and NLRP3 expression. Table 6 dummy alt text NLRP3 expression Chemerin expression Spearman’s rho coefficient test High Low r P High 75 9 0.383 .000 ⁎⁎⁎ Low 34 27 ⁎⁎⁎ P <.001. Expression of chemerin and NLRP3 in OSCC. A, The results indicated that NLRP3 was highly expressed in the high-expression region of chemerin. B, Immunofluorescence double staining showed that chemerin (488 nm) and NLRP3 (650 nm) could be coexpressed and localised in OSCC cells (red for chemerin and green for NLRP3). C, Chemerin overexpression led to the increased expression of pyroptosis-related proteins, and vice versa. When the NLRP3 inhibitor (MCC950) was added to each group, the expression level of pyroptosis-related proteins was decreased. GAPDH was used as a control. Correlation analysis of chemerin expression and NLRP3 expression. ⁎⁎⁎ P <.001. In exploring the effects of tumour pyroptosis on EMT, EMT-related proteins, including E-cadherin, N-cadherin and vimentin, were detected. In Cal27 cells overexpressing the chemerin group (Cal27-LV5+Chemerin), the expression level of N-cadherin and vimentin increased, whereas the expression level of E-cadherin decreased. After adding MCC950, the expression level was found to be reduced. The opposite result was found in SCC15 cells with chemerin knockdown, but adding MCC950 still decreased the expression level ( Figure 7 ). Fig. 7 Chemerin promotes OSCC EMT through pyroptosis. The expression of EMT-related molecules was measured in Cal27 and SCC15 cell lines. After chemerin overexpression, the expression level of N-cadherin and vimentin was increased, and the expression level of E-cadherin was decreased. After MCC950 was added, the expression level of N-cadherin and vimentin was still decreased, whereas the expression level of E-cadherin was increased. Tubulin was used as the control. Fig 7 dummy alt text Chemerin promotes OSCC EMT through pyroptosis. The expression of EMT-related molecules was measured in Cal27 and SCC15 cell lines. After chemerin overexpression, the expression level of N-cadherin and vimentin was increased, and the expression level of E-cadherin was decreased. After MCC950 was added, the expression level of N-cadherin and vimentin was still decreased, whereas the expression level of E-cadherin was increased. Tubulin was used as the control. Transwell experiments were conducted to explore the influence of pyroptosis on the migration and invasion abilities of OSCC cells. The results indicated that the Cal27 cells overexpressing the chemerin group had stronger migration and invasion abilities compared with the control group. On the contrary, the knockdown group (SCC15-Chemerin-shRNA) had fewer migrating and invasive cells than the control group. The number of migrating and invasive cells in each group was significantly reduced with the addition of the NLRP3 inhibitor. The scratch healing experiment had similar experimental results ( Figure 8 ). Fig. 8 Chemerin promotes cell migration and invasion through pyroptosis. A, Transwell migration and invasion experiments showed that the proliferation and migration of OSCC cells were enhanced after chemerin overexpression. After MCC950 was added, the proliferation and migration abilities of cells were weakened. B, Scratch healing experiments showed that OSCC cell migration was enhanced after chemerin overexpression. After adding MCC950, the cell migration ability was weakened. Fig 8 dummy alt text Chemerin promotes cell migration and invasion through pyroptosis. A, Transwell migration and invasion experiments showed that the proliferation and migration of OSCC cells were enhanced after chemerin overexpression. After MCC950 was added, the proliferation and migration abilities of cells were weakened. B, Scratch healing experiments showed that OSCC cell migration was enhanced after chemerin overexpression. After adding MCC950, the cell migration ability was weakened. The effect of pyroptosis on proliferation was detected by the CCK-8 experiment. In addition, the distribution of the cell cycle was detected by flow cytometry, and the expression of cyclinD1 and cyclinE1 was detected by Western blot. The results indicated that the proliferation ability of OSCC cells in the overexpression group was significantly higher than that in the control group. After knockdown, the results were the opposite. The proliferation ability of each group was reduced after adding MCC950 ( Figure 9A ). Second, compared with the Cal27-LV5 group, the Cal27-NC-Chemerin group was significantly enriched in the S phase, and the number of cells in the G1 phase was significantly reduced. On the contrary, the SCC15-NC-Chemerin-shRNA group was significantly more enriched than the SCC15-shRNA group; the cells in the G1 phase were enriched, and the number in the S phase was reduced. After adding MCC950 to each group, the number of S phases decreased in all groups ( Figure 9B ). In this study, the expression of cell cycle proteins such as cyclinD1 and cyclinE1 was detected. The expression level of cyclinD1 and cyclinE1 in the Cal27-NC-Chemerin group was higher than that in the normal group; the opposite was also true. After adding MCC950, this expression level was significantly reduced ( Figure 9C ). Fig. 9 Chemerin promotes cell proliferation through pyroptosis. A, The CCK-8 experiment showed that the OD value of OSCC cells increased after the overexpression of chemerin. The OD value decreases after chemerin knockdown. After MCC950 was added, the OD value of the OSCC cells decreased. B, Flow cytometry results indicated that in the GSDMD-N expression group, the proportion of OSCC cells entering the S phase increased, and cell proliferation was promoted. The knockdown group enriched for S-phase cells and inhibited cell proliferation. C, In the group with high GSDMD-N expression level, the expression level of cyclinE1 and cyclinD1 increased, and after adding MCC950, the expression level of cyclinE1 and cyclinD1 decreased. GAPDH was used as the control. Fig 9 dummy alt text Chemerin promotes cell proliferation through pyroptosis. A, The CCK-8 experiment showed that the OD value of OSCC cells increased after the overexpression of chemerin. The OD value decreases after chemerin knockdown. After MCC950 was added, the OD value of the OSCC cells decreased. B, Flow cytometry results indicated that in the GSDMD-N expression group, the proportion of OSCC cells entering the S phase increased, and cell proliferation was promoted. The knockdown group enriched for S-phase cells and inhibited cell proliferation. C, In the group with high GSDMD-N expression level, the expression level of cyclinE1 and cyclinD1 increased, and after adding MCC950, the expression level of cyclinE1 and cyclinD1 decreased. GAPDH was used as the control. In studying the mechanism by which pyroptosis affects tumour progression, Western blot was used to detect lipid metabolism–related protein expression, including FASN and SREBP. The results indicated that the expression level of FASN and SREBP in the chemerin-overexpressing group (Cal27-LV5-Chemerin) was significantly higher than that in the control group, and vice versa. After adding MCC950, FASN and SREBP were significantly reduced in the overexpression group and knockdown group ( Figure 10 ). Fig. 10 Chemerin affects lipid metabolism through pyroptosis. In both cell lines, the expression level of FASN and SREBP was significantly increased in the GSDMD-N high-expression group, and vice versa. GAPDH was used as the control. Fig 10 dummy alt text Chemerin affects lipid metabolism through pyroptosis. In both cell lines, the expression level of FASN and SREBP was significantly increased in the GSDMD-N high-expression group, and vice versa. GAPDH was used as the control. The results of the nude mouse experiment indicated that the average tumour volume and weight of the Cal27-Chemerin group were 219.33 mm 3 and 0.58 g, respectively, whereas those in the Cal27 group were 41.47 mm 3 and 0.09 g, respectively ( Figure 11 ). By contrast, the tumours in the knockdown group (SCC15-Chemerin-SHRNA: 46.4 mm 3 , 0.15 g) were smaller and lighter than those in the SCC15 group (126.53 mm 3 , 0.33 g). The IHC results indicated that the expression level of pyroptosis-related molecules (GSDMD and IL-1β) and NLRP3 was higher in the chemerin overexpression group (Cal27-Chemerin group) than in the control group, and vice versa. Fig. 11 Effect of chemerin on the formation of subcutaneous tumours in nude mice. A, The results of tumour transplantation in nude mice indicated that the tumours in the CAL27-Chemerin group were larger and heavier than those in the Cal27 group ( P < .05). The tumours in the SCC15 group were larger and heavier than those in the SCC15-Chemerin-shRNA group ( P < .05). B, The immunohistochemical results indicated that the expression level of pyroptosis-related molecules (GSDMD and IL-1β) and NLRP3 was higher in the chemerin overexpression group and lower in the chemerin knockdown group. Fig 11 dummy alt text Effect of chemerin on the formation of subcutaneous tumours in nude mice. A, The results of tumour transplantation in nude mice indicated that the tumours in the CAL27-Chemerin group were larger and heavier than those in the Cal27 group ( P < .05). The tumours in the SCC15 group were larger and heavier than those in the SCC15-Chemerin-shRNA group ( P < .05). B, The immunohistochemical results indicated that the expression level of pyroptosis-related molecules (GSDMD and IL-1β) and NLRP3 was higher in the chemerin overexpression group and lower in the chemerin knockdown group.

Paraffin sections of 145 cases of OSCC tissue and 16 cases of normal mucosal tissue were collected from the Affiliated Hospital of Qingdao University from 2010 to 2020. All patients had complete clinicopathological data. The research protocol was approved by the Ethics Committee of the Medical College of Qingdao University (QDU-HEC-2024257). Informed consent was obtained from all patients participating in this study. The antibodies used in this experiment were as follows: chemerin (ab72965, dilution of 1:100), Gasdermin D (GSDMD) (Cat No. 20770-1-AP, dilution of 1:100), NLRP3 (bs-41293R, dilution of 1:150) and IL-1β ( EPR21086 , dilution of 1:100). All sections were assessed independently by 2 senior pathologists. The scoring criteria for IHC were aligned with methodologies previously documented in the literature, 34 and IHC was scored as follows: Score = staining intensity × percentage of positive cells. Staining intensity was scored as follows: 0, absent; 1, weak expression; 2, moderate expression; and 3, strong expression. The percentage of positive cells was scored as follows: 0 (5%-25%), 1 (5%-25%), 2 (25%-50%), 3 (50%-75%) and 4 (>75%). The abovementioned scores ranged from 0 to 12. A total score of 2 or less was considered negative expression; a total score of > 2 ≤ 6 was considered weak expression; a total score of > 6 ≤ 9 was considered moderate expression; and a total score of > 9 ≤ 12 was considered high expression. In the following analysis we divided them into 2 groups. According to the median value (6) of the total scores, samples were divided into a low-expression group (≤6) and a high-expression group (>6), which is consistent with previous studies in non–small cell lung cancer and bladder cancer. 37 , 38 The human oral squamous cell lines SCC9, SCC15 and CAL27 were purchased from the Chinese Academy of Sciences. Cells were cultured in modified Eagle’s medium containing 10% FBS with 1% penicillin–streptomycin in an incubator at 37 °C and 5% CO 2 . In the experiment, logarithmic cells were collected and experimented. Lentivirus was synthesised by Genepharma located in Shanghai, China. The sequences of the Chemerin-shRNA and scramble controls were as follows: Chemerin-shRNA-512: 5′-GCTTCTACTTCCCTGGACAGT-3′, negative control (Scr-shRNA), 5′-ACGUGACACGUUCGGAGAADTDT-3′. Chemerin was overexpressed by using an overexpression vector plasmid (chemerin). Empty plasmids were used as the negative control (NC). All the primers were synthesised by BGI Gene. The sequences of the primers used in this assay are as follows: Chemerin: 5′-AGACAAGCTGCCGGAAGAGG-3′ (upper) and 5′-TGGAGAAGGCGAACTGTCCA-3′ (lower), GAPDH: 5′-CGGAGTCAACGGATTTTTGTTCGTAT-3′ (upper) and 5′-AGCCTTCTCCATGGTGGTGAAGAC-3′ (lower). Protein preparation and Western blot analysis were conducted following a previously established protocol. 39 The antibodies used in this study, along with their corresponding dilution concentrations, were as follows: GAPDH (Cat No. 60004-1-Ig, ProteinTech, dilution: 1:8000), chemerin (ab72965, Abcam, dilution of 1:500), β-actin (Cat No. 81115-1-RR, ProteinTech, dilution of 1:4000), GSDMD (Cat No. 20770-1-AP, ProteinTech, dilution: 1:5000), GSDMD-N (A18281, abclonal, dilution: 1:1000), NLRP3 (bs-41293R, Bioss, dilution: 1:2000), IL-1β ( EPR21086 , Abcam, dilution: 1:1000),Caspase-1(ET1608-69, ZEN-BIOSCIENCE, 1:5000), Ecadherin (ab314063, Abcam, dilution of 1:500), N-cadherin (EPR1791-4, Abcam, dilution of 1:500), vimentin (EPR3776, Abcam, dilution of 1:500), cyclin D1 (A2708, abclonal, dilution: 1:1000), cyclin E1 (A14225, abclonal, dilution: 1:500), fatty acid synthase (FASN, 200194, ZEN-BIOSCIENCE, dilution of 1:500), and sterol regulatory element-binding protein (SREBP, Cat No. 14088-1-AP, ProteinTech, dilution of 1:4000). The NLRP3 inhibitor (MCC950) was purchased from MCE (Item No. CSN18163 ). The supernatant of the cell culture was collected. The level of IL-1β was measured using an ELISA kit (Cat No. KE00021) according to the manufacturer's instructions. Deparaffinised hydration of paraffin sections and antigen repair were performed as in IHC. The tissues were then blocked with 5% goat serum for 30 min. The blocking solution was discarded, and the tissues were incubated with primary antibodies (Chemerin, ab72965, dilution of 1:100; GSDMD Cat No. 20770-1-AP, dilution of 1:100; NLRP3, bs-41293R, dilution of 1:50) at 4 °C overnight. After performing 3 washes with PBS, each lasting 5 minutes, 2 different fluorescent secondary antibodies were added dropwise to the sections, and the sections were incubated at 37 °C for 2 hours in the dark. The slides were sealed with DAPI. Cells from the logarithmic growth phase were used for the experiments. After washing OSCC cells with PBS, the cells were digested using pancreatic enzymes. After centrifugation for 5 minutes, the cells were fixed overnight in prechilled 70% ethanol at 4 °C. RNase A and iodopropidine were added to the centrifuge tubes and incubated for 30 minutes at 37 °C. Subsequently, the cells were analysed using flow cytometry. Cell migration and invasion assays were performed using a 24-well Transwell chamber equipped with an 8-mm polycarbonate membrane. The middle Transwell plate was added with matrix glue for the invasion test, whereas the upper chamber was not added with matrix glue for the migration test. The treated cells were incubated in a serum-free medium for 24 hours. The lower chamber was filled with 500 mL of a high-concentration serum culture medium, and the upper chamber was filled with a serum-free cell suspension. After incubation at 37 °C for 24-48 hours, the experiment was terminated. The cells subjected to treatment were fixed with formaldehyde for a duration of 30 minutes and subsequently stained with a 0.5% solution of crystal violet. When the cells in the 6-well plate reached around 90%, the monolayer cultured cells were gently scratched by using the new 200 microtips. After gently rinsing with PBS, a serum-free medium was added for subsequent experiments. Microscopic images of the culture plates were recorded at 0 and 24 h and analysed using ImageJ. CCK-8 experiments were performed using 96-well plates. After trypsinisation, the cell suspension was counted on a counting plate. Approximately 2,000 cells were added to each well, and CCK8 was determined at various time points (1, 2, 3, 4 and 5 days). Each well had 10 μL of CCK-8 solution added and then incubated for 2 hours. The absorbance was subsequently measured at 450 nm using a fully automated microplate reader. The strains of BALB/C 4-week-old female nude mice were purchased from Beijing HFK Biotechnology. The nude mice were randomly divided into 4 groups: Cal27, Cal27-Chemerin, SCC15 and SCC15-Chemerin-shRNA. Subcutaneous tumours were established by injecting 1×10⁶ cells into the axillary subcutaneous tissue. During the 60-day period after the injection, the tumour size was measured every 5 days. Tumour volume was determined as V = 0.5 × a × b², where a represents the longest tumour diameter and b represents the shortest tumour diameter. The experiment received approval from the Ethics Committee of the Medical School of Qingdao University (QDU-AEC-2024450). The data were subjected to statistical analysis using the SPSS 25.0 software package. The relationship between chemerin expression and GSDMD was analysed using Spearman’s hierarchy correlation. Chemerin expression and the relationship between GSDMD and clinicopathological factors were tested using the Mann–Whitney test. Survival analysis was conducted using the Kaplan–Meier method and Logrank test, and both univariate and multivariate analyses were executed using the Cox regression model. Western blot used ImageJ to analyse greyscale values. P <.05 indicated significant differences. Each experiment was repeated 3 times.

Pyroptosis can be triggered through 3 different signalling pathways, namely the inflammasome, apoptotic caspase and granzyme pathways. In recent years, the correlation between pyroptosis and tumours has attracted increasing attention. Previous studies showed that Gasdermin protein is aberrantly expressed in tumours and is associated with patient prognosis, 40 , 41 such as breast cancer, 42 non–small cell lung cancer 37 and renal clear cell carcinoma. 43 Our study found that pyroptosis exists in OSCC tissues. IHC results showed that GSDMD and IL-1β were more highly expressed in OSCC compared to normal mucosal tissues. Statistical analysis showed that the high expression level of GSDMD was associated with poor differentiation, a later stage, lymph node metastasis and tumour recurrence. These results indicated that pyroptosis of tumour cells was related to adverse clinical outcomes. Gao et al. studied non–small cell lung cancer 37 and showed that GSDMD was significantly increased, and its high expression level was associated with later stage and large tumours. Xie et al. 43 revealed that GSDMD was also strongly expressed in renal clear cell carcinoma 40 through IHC experiments and was significantly correlated with TNM stage, grade and lymph node metastasis. In addition, previous studies revealed that the expression level of GSDMB in breast cancer 42 was significantly higher than that in normal tissues, and its high expression level is associated with high metastasis rate and low survival rate. In the study of hepatocellular carcinoma, 44 GSDME significantly increased, and it was associated with later stages. Kaplan–Meier survival analysis was used to identify patients with high GSDMD expression levels and short OS and DFS time. Cox regression analysis showed that the expression of GSDMD was an independent prognostic indicator for patients with OSCC. This finding indicates that the pyroptosis of OSCC cells was associated with poor prognosis. Similar results have been observed in other tumours. For example, the high expression level of GSDMD in renal clear cell carcinoma 43 promoted the release of IL-1β, further accelerating the proliferation of renal cell cancer cells and shortening the OS and DFS time. In non–small cell lung cancer, 37 the high expression level of GSDMD was associated with low survival rate, and knocking down its expression inhibits the EGFR/Akt pathway and tumour proliferation. Similar result was also found in pancreatic cancer, 45 where overexpression of GSDMC increased the degree of pyroptosis in tumour cells, shortened OS time and was associated with drug resistance to chemotherapy. Their study also showed that pyroptosis can promote cancer cell proliferation and accelerate tumour progression through the pentose phosphate pathway. These studies indicated that pyroptosis of OSCC tissues may also promote tumour progression through certain pathways. The effect of pyroptosis on tumour biological behaviour was further explored through in vitro experiments, including Transwell, scratch healing, CCK8, cell cycle and EMT assays. The results indicated that the migration and invasion abilities of OSCC cells in the group with a high expression level of GSDMD-N were more enhanced than those in the group with a low expression level of GSDMD-N, and vice versa. The Western blot results indicated that the increasement of pyroptosis can promote the EMT of tumour cells. Therefore, after tumour pyroptosis, the adhesion of tumour cells was reduced because of EMT, which promotes the invasion and migration of tumour cells. Previous studies on endometriosis 46 showed that prostaglandin E2 induces the NLRP3/caspase-1 apoptotic pathway in cells to upregulate the expression level of HMGB1, vimentin and E-cadherin, enhancing cell migration. Other studies also revealed that HMGB1 released after pyroptosis can promote the occurrence and development of tumours by participating in the ERK1/2, EMT and Wnt pathways. 47 , 48 A study on bladder cancer revealed that GSDMB expression was elevated in tumours, 49 promoting pyroptosis and interacting with STAT3 to activate phosphorylation, which led to the growth and invasion of tumour cells. Our results also indicated that pyroptosis promoted cell proliferation. The effects of pyroptosis on cell proliferation were further investigated. The Western blot results indicated that the expression of cyclinD1 and cyclinE1 was upregulated after pyroptosis enhancement. In addition, the results of cell cycle flow cytometry indicated that pyroptosis enhancement could increase the proportion of OSCC cells entering the S phase. Previous studies have demonstrated that following pyroptosis, tumour cells release a large number of proinflammatory cytokines (such as IL-1β, IL-18 and TNF-α), chemokines (such as CCL2 and CXCL8) and damage-associated molecular patterns (including ATP and HMGB1) into the tumour microenvironment (TME). 50 These factors act on surviving tumour cells in a paracrine manner via their corresponding cell surface receptors and activate downstream signalling pathways including NF-κB and PI3K/Akt. 51 , 52 These activated pathways directly upregulate the expression of cell cycle–related proteins (e.g. Cyclin D1, CDK4/6 and c-Myc) and inhibit the expression of cell cycle checkpoint proteins (e.g. p21 and p27), thereby accelerating the G1/S phase transition and cell cycle progression in surviving tumour cells. 53 , 54 Furthermore, the inflammatory factors released during pyroptosis will also recruit immunosuppressive cells (M2-type macrophages and regulatory T cells) to the tumour microenvironment (TME), which further suppresses the antitumour immune response and creates a proliferative microenvironment for the surviving tumour cells. 55 , 56 Therefore, the pyroptosis of OSCC cells may promote tumour proliferation by regulating the cell cycle. A previous study on nonalcoholic fatty liver disease 57 found that high-fat diet can increase the number of pyroptosis cells and promote the accumulation of lipoproteins, including FASN, SREBP and fatty acid–binding protein (FABP4), which implies the possible connection between pyroptosis and lipid metabolism. Therefore, we further examined whether pyroptosis in OSCC affects lipid metabolism. The Western blot results indicated that the expression level of the lipid metabolism–related proteins FASN and SREBP was upregulated with a higher degree of pyroptosis. In addition, our study also found that NLRP3 expression facilitated the activation of caspase-1. Studies in hepatocytes further showed that caspase-1 activation induces SREBPs activation mediated by site-1 protease (s1p) and site-2 protease (S2P), which in turn upregulates the expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) and FASN—the target genes of SREBP-2 and SREBP-1c, respectively. 58 , 59 Therefore, we speculate that pyroptosis of OSCC cells may regulate lipid metabolism through caspase-1, which needs further exploration. Previous studies showed that many factors can promote cell pyroptosis, including ubiquitin-specific protease 48 (USP48), 60 17β-estradiol, lipopolysaccharide (LPS) 61 and 3-domain protein 21 (TRIM21). 62 For example, in diabetic cardiomyocytes, 61 LPS activated NLRP3-induced pyroptosis by upregulating ROS. A study also showed that TRIM21 62 can bind to GSDMD through its structural domain (PRY-SPRY) and promote GSDMD-N aggregation, thereby regulating pyroptosis. Moreover, a previous study on diabetes indicated that chemerin acts on its receptors to promote pyroptosis in adipocytes. Our previous study showed that chemerin was highly expressed in OSCC and was associated with poor clinical outcomes. 34 , 39 Therefore, we investigated the effect of chemerin expression on the pyroptosis of OSCC cells. Our IHC results indicated that the expression of GSDMD was positively correlated with chemerin (r = 0.229, P = .006). Chemerin and GSDMD were also colocalised in OSCC cells. In in vitro experiments, Western blot results indicated that chemerin expression promotes the upregulation of GSDMD-N, GSDMD and IL-1β expression. In addition, the high expression level of chemerin and GSDMD was associated with lower differentiation, a later stage, lymph node metastasis and tumour recurrence. Survival analysis results indicated that patients with both high expression levels had shorter OS and DFS time than the other 2 groups. In addition, in nude mouse experiments, the tumours in the chemerin overexpression group were larger and heavier with a higher expression level of GSDMD and IL-β than those in the control group, whereas the results in the chemerin knockdown group were the opposite. These results indicate that chemerin expression in OSCC may promote the pyroptosis of tumour cells and lead to the development of tumours. We further explored the possible molecular pathway by which chemerin expression promotes the pyroptosis of tumour cells. Previous studies showed that inflammasome plays a key role in the process of pyroptosis. The inflammasome is a protein complex composed mainly of receptor proteins, adaptor proteins (ASC) and downstream caspase families. The main function of the inflammasome is to recruit and activate pro-caspase-1, which promotes the maturation of pro-IL-1β and pro-IL-18, thereby triggering pyroptosis. 63 A previous study on OSCC tissues 35 showed that the expression level of NLRP3 was significantly increased and related to poor prognosis. Studies on preeclampsia 36 indicated that chemerin expression can promote the AMPK/TXNIP/NLRP3 pathway and aggravate the severity of preeclampsia. Therefore, we determined whether chemerin may promote pyroptosis of tumour cells through the NLRP3 pathway in OSCC. Our IHC results indicated that chemerin expression was positively correlated with NLRP3 (r = 0.383, P < .001). In addition, chemerin expression can upregulate the expression level of NLRP3 in in vitro experiments. In verifying the NLRP3 pathway, its inhibitor (MCC950) was used, and the aforementioned experiments were repeated. In addition, the results indicated that when MCC950 was added to all groups, the expression of GSDMD-N was lower, and the proliferation, migration and invasion of OSCC cells were weakened. Moreover, fewer OSCC cells entered the S phase, and the expression level of lipid-related proteins was decreased. Therefore, chemerin exerts its effect on the pyroptosis of tumour cells mainly through NLRP3. Overall, our results indicated that the high expression level of GSDMD in OSCC was associated with a poor prognosis for patients. The expression of chemerin can promote the pyroptosis of OSCC cells through NLRP3/GSDMD-N and accelerate tumour development through the regulation of cell cycle, EMT and lipid metabolism. Chemerin and its mediated NLRP3/GSDMD-N pathway provide a new possible target for OSCC’s treatment. Inhibiting the expression of chemerin or blocking the NLRP3/GSDMD-N pathway may effectively restrict OSCC’s progression to improve OSCC patients’ clinical outcomes and prognosis.

In summary, we revealed that the expression of chemerin can regulate the NLRP3/GSDMD-N pathway to promote tumour progression. However, this study did not elaborate on the regulation between pyroptosis and lipid metabolism. Furthermore, the detection of GSDMD-mediated pyroptosis (e.g. IHC staining for GSDMD-N and measurement of inflammatory cytokine secretion) was performed at fixed time points in in vitro cell models and clinical tissue samples, which reflects only a transient state of the dynamic pyroptosis process rather than its continuous and dynamic changes during tumour progression. Pyroptosis is a spatiotemporally regulated cellular process: its initiation, execution and subsequent inflammatory factor release exhibit distinct kinetic characteristics in response to different stimuli (e.g. oxidative stress, immune cell attack) and at different stages of tumour development. Our fixed-time-point detection cannot fully reveal the temporal and spatial dynamics of GSDMD activation, as well as the subsequent paracrine regulation of surviving tumour cells in the tumour microenvironment. In addition, the heterogeneity of tumour cells and the complexity of the tumour microenvironment lead to the asynchronous occurrence of pyroptosis in different tumour cell subpopulations, which makes the snapshot data unable to reflect on the pyroptosis status of the entire tumour tissue comprehensively. In the future, we will continue to investigate the relationship between pyroptosis and lipid metabolism, as well as their potential underlying mechanisms.

Oral squamous cell carcinoma (OSCC) is a malignant tumour that arises in the oral mucosa, including the tongue, gingiva and floor of the mouth. OSCC is a prevalent malignant tumour of the head and neck. 1 Its pathogenesis is complex, 2 which relates to various factors. 3 , 4 Because of OSCC’s high risk of recurrence and metastasis, the affected patients continue to have an undesirable survival rate despite the several treatments that have been established to date. 5 , 6 Therefore, exploring the molecular mechanism underlying the progression and development of OSCC is necessary to improve its poor clinical outcomes. In contrast to necrosis and apoptosis, pyroptosis is a type of planned cell death that is accompanied by inflammatory reactions. 7 This process is characterised by the cleavage of the gasdermin family proteins. 8 These proteins can be cleaved into a C-terminal domain and an active N-terminal domain by upstream signalling molecules, 9 , 10 forming pores in the cell membrane, leading to the formation of osmotic pressure differences between the inside and outside of the cell as well as the release of IL-1β, IL-18, High Mobility Group Box 1 protein (HMGB1), and ATP, among other factors. 11 Gram-negative bacteria were first found to induce caspase-1-dependent apoptosis in infected macrophages, 12 and the pathway was termed ‘pyroptosis’ in 2001. 13 Previous studies on pyroptosis focused on diseases such as diabetes, 14 , 15 sepsis, 16 , 17 , 18 and atherosclerosis. 19 , 20 For example, in diabetic mice, 21 the induction of high-fat diet or streptozocin can increase the expression level of IL-1β and caspase-1, activate the pyroptosis of pancreatic β-cells, 22 and facilitate the onset and progression of diabetes. In recent years, a growing body of research has shown that pyroptosis may play a role in tumour progression, but its specific mechanisms may be complex. For one thing, various inflammatory mediators released during pyroptosis create a suitable tumour microenvironment for the survival of cancer cells. 23 , 24 For example, research in pancreatic cancer 23 showed that the cleavage of the gasdermin protein by caspase-3 induces the transition from apoptosis to pyroptosis of pancreatic tumour cells and releases a variety of cytokines to enhance its survivability. For another, it has the potential to inhibit tumour cells by eliciting an antitumour immune response. 25 , 26 For example, 17β-estradiol (E2) activates inflammation-induced pyroptosis in hepatocellular carcinoma 25 and inhibits tumour progression through the estrogen receptor beta (ERβ)/5′ AMP-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR) signalling pathway. To date, relatively limited studies on pyroptosis in OSCC are available and the existing reports mainly focus on its correlation with chemotherapy drug resistance. 27 , 28 The impact of pyroptosis on the prognosis and behaviour of OSCC has remained ambiguous. Pyroptosis may regulate tumour progression and the prognostic outcome in OSCC patients through multiple potential mechanisms. For example, excessive activation of pyroptosis may induce a chronic inflammatory response in the tumour microenvironment, promoting the invasion and lymph node metastasis of OSCC cells by releasing proinflammatory factors such as IL-1β and HMGB1, and thus lead to a poorer prognosis. In addition, the proinflammatory factors released by pyroptosis may recruit a large number of tumour-associated macrophages (TAM), regulatory T cells (Treg) and other immunosuppressive cells to the tumour tissue. These cells may secrete anti-inflammatory factors, inhibiting the activity of cytotoxic T cells and forming an immunosuppressive microenvironment, thus enabling tumour cells to achieve immune evasion. 29 Previous studies have indicated that chemerin expression can promote pyroptosis in diabetes mellitus, 30 but its specific mechanism remains unclear. Chemerin is a fat-related factor with multiple functions, 31 which plays key roles in immune and inflammatory responses as well as glucose and lipid metabolism. It is also implicated in various types of tumours. A study in ovarian cancer showed that overexpression of chemerin upregulated PD-L1, promoting the proliferation and metastasis of cancer cells. 32 The cancer-associated myofibroblast of oesophageal squamous cell carcinoma (OSC) secreted chemerin, which binds to chemR23 on the surface of OSC cells, activating the downstream protein kinase C (PKC)/mitogen-activated protein kinase (MAPK) signalling pathway, inducing the expression and activation of MMP-1/2/3 and ultimately promoting the migration, invasion and proliferation of OSC cells. 33 Its high expression may promote inflammatory and cancer-related pathways, leading to the invasion and metastasis of cancer cells. Our previous study also demonstrated that the expression level of chemerin in OSCC was higher than that in paracancerous tissues, and this elevation was correlated with a poor clinical prognosis. 34 However, the relationship between pyroptosis and chemerin expression was unclear. Previous studies have shown that inflammasome plays a crucial role in the process of pyroptosis. A study on OSCC 35 found that the expression level of the NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) inflammasome was significantly increased in OSCC. In addition, a study on preeclampsia 36 revealed that elevated chemerin expression level can promote the AMPK/Thioredoxin-interacting protein (TXNIP)/ NLRP3 pathway. Therefore, this study further explored whether the expression of chemerin in OSCC can affect the pyroptosis of tumour cells through NLRP3.

None disclosed.