探讨曲马多对宫颈癌细胞增殖与迁移及侵袭的影响及其对Wnt信号通路的调控作用

Tramadol inhibits proliferation,migration and invasion of cervical cancer cells by regulating Wnt signaling pathway

目的曲马多(tramadol)具有抗癌作用,但其对宫颈癌细胞生物学行为的影响尚未阐明,因而本研究探讨曲马多对宫颈癌细胞增殖、迁移及侵袭的影响及其对Wnt信号通路的调控作用。方法采用甲基噻唑基四唑(MTT)实验选择适宜的曲马多浓度并分析其对正常宫颈细胞Ect1/E6E7、宫颈癌Ca Ski细胞增殖的影响;Transwell小室实验检测曲马多曲马多对Ca Ski细胞迁移及侵袭能力的影响;分别加入Wnt信号通路抑制剂DK419与激活剂SKL2001处理Ca Ski细胞,观察细胞增殖、迁移及侵袭能力变化。蛋白免疫印迹法(Western blot)检测Wnt1、细胞周期蛋白1(Cyclin D1)、β-连环素(β-catenin)、上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)的表达水平,进一步证实曲马多对细胞增殖、迁移及侵袭的作用。结果曲马多能显著抑制Ca Ski细胞增殖、迁移及侵袭(P<0.05),抑制神经型钙黏蛋白的表达(P<0.05),促进上皮型钙黏蛋白的表达(P<0.05),且呈剂量依赖效应;曲马多可降低Wnt1、Cyclin D1、β-catenin的表达(P<0.05);抑制Wnt信号通路后,CaSki细胞存活率显著降低(P<0.05),迁移细胞数与侵袭细胞数减少(P<0.05),神经型钙黏蛋白表达下调(P<0.05),上皮型钙黏蛋白表达上调(P<0.05);激活Wnt信号通路能够明显逆转曲马多对Ca Ski细胞增殖、迁移及侵袭的抑制作用(P<0.05)。结论曲马多可抑制宫颈癌细胞增殖、迁移及侵袭,其机制可能是通过抑制Wnt信号通路而发挥作用。

Objective Tramadol had an anticancer effect,but its effect on the biological behavior of cervical cancer cells has not been elucidated.Therefore,this study explored the effect of tramadol on the proliferation,migration and invasion of cervical cancer cells and its regulation on Wnt signaling pathway.Method The appropriate tramatol concentration was selected by MTT assay and its effect on the proliferation of normal cervical cells Ect1/E6 E7 and cervical cancer CaSki cells was analyzed.The Transwell chamber assay was used to detect the effect of tramadol on the migration and invasion of CaSki cells.CaSki cells were treated with Wnt signaling pathway inhibitor DK419 and activator SKL2001,respectively,to observe changes in cell proliferation,migration and invasion.The expression levels of Wnt1,Cyclin D1,β-catenin,E-cadherin and N-cadherin were detected by Western blot,further confirmed the effect of tramadol on cell proliferation,migration and invasion.Results Tramadol significantly inhibited the proliferation,migration and invasion of CaSki cells(P<0.05),inhibited the expression of N-cadherin(P<0.05),and promoted the expression of E-cadherin(P<0.05)in a dose-dependent manner.Tramadol reduced the expression of Wnt1,Cyclin D1,andβ-catenin(P<0.05).After inhibiting Wnt signaling pathway,the survival rate of CaSki cells was significantly decreased(P<0.05),the number of migrated cells and the number of invasive cells decreased(P<0.05),the expression of N-cadherin was down-regulated(P<0.05),and the expression of E-cadherin was up-regulated(P<0.05).Activation of Wnt signaling pathway significantly reversed the inhibitory effect of tramadol on proliferation,migration and invasion of CaSki cells(P<0.05).Conclusion Tramadol inhibits the proliferation,migration and invasion of cervical cancer cells,and its mechanism may be through inhibition of Wnt signaling pathway.