大豆苷元靶向MMP12发挥抗口腔鳞状细胞癌的作用

Daidzein targeting MMP12 against oral squamous cell carcinoma

目的分析大豆苷元靶向MMP12发挥抗口腔鳞状细胞癌的作用。方法数据库分析大豆苷元靶点及其与口腔鳞状细胞癌病理特征的关系。将CAL-27和Tca8113细胞分为对照组、大豆苷元组、大豆苷元+MMP12组。MTT实验检测细胞存活率,CCK-8实验检测细胞增殖活性,克隆形成实验检测细胞生长情况,Transwell实验检测细胞迁移和侵袭情况,Western blot检测MMP12蛋白表达。裸鼠皮下注射人口腔鳞状细胞癌CAL-27细胞株构建皮下移植瘤模型,daidzein组每隔1 d注射大豆苷元1次,control组注射等量生理盐水。计算肿瘤体积和质量,免疫组化法检测移植瘤组织中Ki67蛋白表达。结果GEO2R分析显示,MMP12在口腔鳞状细胞癌中表达(P<0.05),且MMP12表达与淋巴结转移相关(P<0.05)。GSEA分析表明,与细胞增殖、迁移和侵袭相关的基因集均富集在MMP12高表达组。大豆苷元浓度越高,作用时间越长,口腔鳞状细胞癌细胞活性、克隆细胞数、细胞侵袭数和迁移数明显越低/少(P<0.05)。daidzein组移植瘤体积、质量、Ki67蛋白表达水平均明显小/低于control组(P<0.05)。随着大豆苷元作用时间延长或浓度升高,MMP12表达水平明显下降(P<0.05)。与对照组相比,大豆苷元组口腔鳞状细胞癌细胞MMP12表达水平、细胞活性、克隆细胞数、细胞迁移和侵袭数明显降低/减少(P<0.05);与大豆苷元组比较,大豆苷元+MMP12组口腔鳞状细胞癌细胞活性、克隆细胞数、细胞迁移和侵袭数明显升高/增加(P<0.05)。结论大豆苷元可通过靶向MMP12下调MMP12表达,进一步抑制口腔鳞状细胞癌细胞增殖、迁移和侵袭,从而发挥抗肿瘤作用。

Objective To analyze the prevention effect of daidzein on oral squamous cell carcinoma by targeting MMP12.Methods The targets of daidzein and their relationship with pathological characteristics of oral squamous cell carcinoma were analyzed by databases.CAL-27 and Tca8113 cells were divided into the control group,the daidzein group and the daidzein+MMP12 group.The cell survival rate was determined by the MTT assay,the cell proliferative activity was determined by the CCK-8 assay,the cell growth was determined by the clone formation assay,the cell migration and invasion were determined by the Transwell assay,and the MMP12 protein expression was determined by Western blot.The nude mice were given subcutaneous injection of human oral squamous cell carcinoma CAL-27 cell line to construct the subcutaneous xenograft tumors models.The mice in the daidzein group were injected with daidzein once every 1 day,while the mice in the control group was injected with the same volume of normal saline.The tumor volume and mass were calculated,and the expression of Ki67 protein in the xenografts tissues was detected by immunohistochemistry.Results GEO2R analysis showed that MMP12 was expressed in oral squamous cell carcinoma(P<0.05),and the expression of MMP12 was related to lymph node metastasis(P<0.05).GSEA analysis showed that the gene sets related to cell proliferation,migration,and invasion were all enriched in the MMP12 high expression group.The higher the concentration of daidzein,the longer the action time,the lower/fewer the cell activity,the number of cloned cells,the number of cell invasion and migration of the oral squamous cell carcinoma(P<0.05).The volume,mass and Ki67 protein expression level of xenografts in the daidzein group were significantly smaller/lower than those in the control group(P<0.05).With the prolongation of daidzein action time or the increase of daidzein concentration,the expression level of MMP12 was significantly decreased(P<0.05).Compared with the control group,the expression level of MMP12,the cell activity,the number of cloned cells,the number of cell invasion and migration of oral squamous cell carcinoma cell in the daidzin group were significantly reduced(P<0.05).Compared with the daidzein group,the cell activity,the number of cloned cells,the number of cell migration and invasion of oral squamous cell carcinoma cell in the daidzein+MMP12 group were significantly increased(P<0.05).Conclusion Daidzein can further inhibit the proliferation,migration and invasion of oral squamous cell carcinoma by targeting MMP12 to down-regulate the expression of MMP12,thereby exerting the anti-tumor effect.