波形蛋白促进激素非依赖性前列腺癌细胞侵袭与转移的机制探讨

Studies on the mechanism underlying androgen-independent prostate cancer invasion and metastasis promoted by vimentin

目的:探讨波形蛋白促进激素非依赖性前列腺癌细胞侵袭与转移的分子机制。方法:应用波形蛋白特异性扩增引物,借助RT-PCR获得波形蛋白正义全长和反义全长cDNA片段,分别插入pcDNA3.1(+)和pcDNA3.1(-)表达载体中,并将重组质粒导入激素非依赖性前列腺癌配对细胞系中,G418稳定筛选;应用体外运动和侵袭试验比较细胞侵袭能力的改变;用蛋白免疫印迹法检测相关蛋白的表达。结果:波形蛋白通过C-src激酶调节E-钙黏蛋白(E-cadherin)/β-连环蛋白(β-cate-nin)复合物,促进激素非依赖性前列腺癌细胞侵袭与转移。结论:波形蛋白可以作为判断激素非依赖性前列腺癌细胞侵袭与转移的一个重要指标。

Objective ：To study the molecular mechanism underlying androgen-independent prostate cancer invasion and metastasis promoted by vimentin. Methods： With specific amplified primer of vimentin,full-length vimentin and full-length antisense-vimentin cDNA fragments were synthesized by reverse transcriptase polymerase chain reaction （RT-PCR）. RT-PCR products were cloned into plasmid pcDNA3.1 （ ＋ ） and pcDNA3.1 （ - ）. The recombinant vimentin plasmids were transfected into androgen-independent prostate cancer PC-3M-2B4 and PC-3M-1 E8 cells. The positive clones were screened by G418. The transwell chamber migration assay was used to study cell invasion, Western blotting was used to determine specific protein expression in the cells. Results： Vimentin promoted androgen-independent prostate cancer cell invasion and metastasis by regulating the E-cadherin/β-catenin complex via C-src kinase. Conclusion： Vimentin could be used as an important prognostic factor for androgen-independent prostate cancer cell invasion and metastasis.