靶向沉默维生素D受体基因对前列腺癌细胞迁移及侵袭性的影响

Effect of silencing the VDR gene on the migration and invasion of prostate cancer cells

目的:探讨小干扰RNA靶向沉默维生素D受体(VDR)对前列腺癌PC-3细胞生物学行为的影响。方法:构建VDR-shRNA慢病毒载体,用RT-PCR和Western印迹法分别检测VDR的mRNA和蛋白表达水平;通过细胞划痕愈合实验和Transwell小室检测VDR被沉默后的PC-3细胞迁移能力和侵袭性的变化。结果:VDR-shRNA质粒显著干扰VDR表达,并成功筛选VDR-shRNA干扰稳定的细胞株;细胞划痕实验结果显示划痕愈合率VDR干扰组为59%明显低于空白对照组73.6%和LV3阴性对照组的77.8%,组间差异有显著性(P<0.05),空白对照组和LV3阴性对照组组间差异无显著性(P>0.05)。Transwell小室实验显示VDR干扰组透膜细胞数量明显低于空白对照组和LV3阴性对照组约为50%,细胞组间差异有显著性(P<0.05),空白对照组和LV3阴性对照组组间差异无显著性(P>0.05)。VDR干扰组细胞的迁移和侵袭能力明显低于对照组细胞。结论:VDR基因表达水平能影响前列腺癌细胞的迁移及侵袭能力,VDR低表达可致前列腺癌细胞迁移及侵袭能力下降。

Objective ： To investigate the effect of small interfering RNA silencing the vitamin D receptor （VDR） on the biologi- cal behavior of prostate cancer PC-3 cells. Methods ： We constructed the VDR-shRNA lentiviral vector and determined the mRNA and protein expressions of VDR by RT-PCR and Western blot. Using scratch wound healing and Transwell chamber assays, we detected the changes in the migration and invasiveness of the PC-3 cells after silencing VDR. Results ： The VDR-shRNA plasmid significantly interfered the VDR expression and successfully screened the cell lines with stable VDR-shRNA interference. The rate of scratch wound healing was markedly lower in the VDR interference group than in the blank control and LV3 negative control groups （59% vs 73.6% and 77.8% , P 〈0.05） , but with no statistically significant difference between the latter two （P 〉0.05） , and so was the count of permeable cells （ P 〈 0.05 ）, but with no significant difference between the latter two groups, either （ P 〉 0.05 ）. The migration abili- ty and invasiveness of the VDR-treated cells were remarkably decreased as compared with those of the control cells. Conclusion： Down-regulated expression of the VDR gene may reduce the migration and invasiveness of prostate cancer cells.