周期蛋白依赖性激酶5调节去势抵抗性前列腺癌细胞侵袭转移的机制研究

Mechanism of cyclin dependent kinase 5 in regulating the invasion and metastasis of castration resistant prostate cancer cells

目的研究周期蛋白依赖性激酶5(CDK5)对去势抵抗性前列腺癌细胞DU145生长及侵袭转移的影响,并初步探究其作用机制。方法采用CDK5 siRNA干扰CDK5在DU145细胞内的表达,CDK5抑制剂20-223抑制CDK5在DU145细胞内的活性;采用噻唑蓝(MTT)检测细胞生长活力,单克隆形成实验测定细胞增值能力;采用膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)和碘化丙啶(PI)双染法测定细胞凋亡;划痕实验和Transwell实验测定细胞的迁移能力;Western Blot测定Myc促癌基因(c-Myc)、SRY-box转录因子4(SOX4)及侵袭转移相关蛋白的表达情况。结果抑制CDK5在DU145细胞内的表达和活性,可抑制肿瘤细胞的恶性增殖和侵袭转移(P<0.05),促进肿瘤细胞凋亡(P<0.05),抑制c-Myc、SOX4、波形蛋白(Vimentin)和锌指E-box结合同源盒1(Zeb1)的表达量(P<0.05),同时促进E-钙黏蛋白(E-cadherin)的表达(P<0.05)。结论CDK5表达和活性的降低可通过c-Myc/SOX4/Zeb1信号通路抑制去势抵抗性前列腺癌细胞DU145的恶性增殖和侵袭转移。

Objective To investigate the effects of cyclin dependent kinase 5(CDK5)on the proliferation and metastasis of castration resistant prostate cancer DU145 cells and the corresponding mechanism.Methods CDK5 siRNA was used to interfere with the expression of CDK5 in DU145 cells,and CDK5 inhibitor 20-223 inhibited the activity of CDK5 in DU145 cells.The growth activity of the cells was detected by MTT,and the proliferation ability of the cells was determined by monoclonal formation assay.Apoptosis was determined by Annexin V-FITC and PI double staining.The migration ability of cells was determined by scratch assay and Transwell assay.The expression of Myc proto-oncogene(c-Myc),SRY-box transcription factor 4(SOX4)and metastasis related proteins were determined by Western Blot.Results Inhibition of CDK5 expression and activity in DU145 cells could inhibit the malignant proliferation,invasion and metastasis of tumor cells(P<0.05)and promote the apoptosis of tumor cells(P<0.05),and inhibit the expression levels of C-Myc,SOX4,Vimentin and Zeb1(P<0.05),and promote the expression of E-cadherin(P<0.05).Conclusions The decrease of CDK5 expression and activity can inhibit the malignant proliferation,invasion and metastasis of castration resistant prostate cancer cell DU145 through c-Myc/SOX4/Zeb1 signaling pathway.