小干扰RNA介导的蛋白激酶Cε基因沉默对肾透明细胞癌769P细胞增殖及迁移侵袭的影响

Effects of small interfering RNA against protein kinase CE gene on proliferation, migration and in- vasion of human renal clear cell carcinoma cell line 769P

目的观察下调蛋白激酶Cε（PKCε）基因对。肾透明细胞癌细胞株769P细胞的生长、侵袭及迁移能力的影响。方法用脂质体将PKCε小干扰RNA（siRNA）转染至769P细胞中，逆转录-聚合酶链反应（RT-PCR）及Westernblot分析PKCε基因及蛋白的表达；噻唑蓝（MTT）及单细胞克隆试验测定细胞的生长；划痕及侵袭试验检测细胞的侵袭和迁移能力。结果通过siRNA下调769P细胞中PKCε的表达后，细胞的增殖能力下降明显（P〈0．05）；同时，细胞的侵袭和迁移能力较正常细胞降低，差异有统计学意义（P〈0．05）。结论PKCε可影响人肾透明细胞癌细胞株769P细胞的生长及细胞侵袭、迁移能力。

Objective To evaluate the effects of small interfering RNA （siRNA） against protein kinase C~ （PKCa） gene on the proliferation, migration and invasion of human clear cell RCC cell line 769P cells. Methods FKCε-siRNA was transfected into the 769P cells. The mRNA and protein expres- sion of PKCε was detected by using reverse transcription polymerase chain reaction （RT-PCR） and West- ern blotting, respectively. The proliferation of 769P cells was measured by methyl thiazol tetrazolium （MTT） assay and colony formation assay. Cell migration and invasion was assessed by using wound-healing assay and CHEMICON cell invasion assay, respectively. Results PKCε-siRNA efficiently down-regulated the PKCε expression, which resulted in inhibition of the proliferation, migration, and invasion of clear cell RCC cell line 769P. Conclusion PKCais may be related with proliferation, migration and invasion of 769P cells, and may be a potential therapeutic target for clear cell RCC.