TRPV4通过调控MAPK/ERK信号通路促进卵巢癌细胞的增殖和迁移

TRPV4 promotes the proliferation and migration of ovarian cancer cells by regulating MAPK/ERK signaling pathway

目的:探讨TRPV4通过影响MAPK/ERK信号通路调控卵巢癌细胞的增殖与迁移。方法:RT-qPCR和Western Blot方法检测TRPV4在卵巢癌细胞系和正常卵巢上皮细胞中的表达差异;GEPIA在线数据库分析卵巢癌患者TRPV4表达水平和预后关系;敲低TRPV4,通过RT-qPCR和Western Blot检测转染效率;通过细胞计数试剂盒(CCK8)、克隆形成实验以及Transwell迁移实验检测TRPV4在卵巢癌发生发展过程中增殖及迁移作用;通过GSEA软件富集分析信号通路并用Western Blot验证卵巢癌细胞MAPK/ERK信号通路表达量变化。结果:在卵巢癌细胞及临床组织样本中,TRPV4表达量明显增高,TRPV4表达量较高的患者预后更差。siRNA转染卵巢癌HO8910和Caov3细胞后,与Control组相比,TRPV4 mRNA和蛋白表达显著降低;细胞增殖迁移能力显著减弱;WB实验结果显示ERK总量不变,磷酸化水平降低,进一步实验中证实:敲低TRPV4后,ERK信号通路中的关键蛋白(P90RSK、MEK1/2、MSK1)磷酸化水平降低。结论:TRPV4在卵巢癌细胞及组织中高表达,可能通过正向调控MAPK/ERK信号通路,促进卵巢癌细胞增殖迁移等功能。

Objective:To explore the effect of TRPV4 on the proliferation and migration of ovarian cancer cells by affecting MAPK/ERK signal pathway.Methods:RT-qPCR and Western Blot methods were used to detect the difference of TRPV4 expression between ovarian cancer cell lines and normal ovarian epithelial cells.GEPIA online database was used to analyze the relationship between TRPV4 expression and prognosis in ovarian cancer patients.TRPV4 was knocked down,RT-qPCR and Western Blot were used to detect the transfection efficiency.The proliferation and migration of TRPV4 during the occurrence and development of ovarian cancer were detected by cell counting Kit(CCK8),clone formation assay and Transwell migration assay.The signal pathway was enriched and analyzed by GSEA software,and the expression of MAPK/ERK signal pathway in ovarian cancer cells was verified by Western blot.Results:The expression of TRPV4 was significantly increased in ovarian cancer cells and clinical tissue samples,and the prognosis of patients with high expression of TRPV4 was worse.After transfection of siRNA into ovarian cancer cells(HO8910,Caov3),the expression of TRPV4 mRNA and protein decreased significantly compared with the control group.The ability of cell proliferation and migration decreased significantly.The results of WB experiment showed that the total amount of ERK remained unchanged and the phosphorylation level decreased.Further experiments confirmed that,knocking down TRPV4,the phosphorylation level of key proteins(P90RSK,MEK1/2,MSK1)in the ERK signaling pathway decreased.Conclusion:TRPV4 is highly expressed in ovarian cancer cells and tissues,which may promote the proliferation and migration of ovarian cancer cells by positively regulating MAPK/ERK signal pathway.