沉默PDK1基因抑制卵巢癌细胞系SKOV3的增殖及侵袭

Silencing PDK1 gene inhibits proliferation and invasion of ovarian cancer cell line SKOV3

目的研究小干扰RNA(siRNA)沉默丙酮酸脱氢酶激酶1(PDK1)基因对卵巢癌细胞系SKOV3增殖、迁移、侵袭以及克隆形成能力的影响,并初步探讨其机制。方法采用PDK1 siRNA及control siRNA分别瞬时转染干扰实验组和阴性对照组SKOV3细胞。运用Western blot及real-time PCR验证转染效率后,CCK8法、平板克隆形成实验检测细胞增殖及克隆能力;Transwell小室法及划痕实验检测细胞侵袭及迁移能力;流式细胞计量术探索细胞周期及凋亡情况。结果实验组细胞增殖及克隆(P<0.01),侵袭及迁移能力(P<0.05)显著低于对照组;SKOV3细胞内PDK1基因沉默后缩短了细胞复制期并提高了其早期凋亡能力(P<0.01)。结论沉默PDK1基因可显著抑制SKOV3细胞的增殖、迁移、侵袭以及克隆形成能力,并可缩短细胞S期,增加细胞凋亡率。

Objective To investigate the effects of small interfering RNA mediated pyruvate dehydrogenase kinase 1 silencing on the proliferation, migration, invasion and clonogenic ability of SKOV3 cells and preliminarily discuss its mechanism. Methods Transient transfection interfered with SKOV3 cells in both the experimental group and the control group. Western blot and real-time PCR were used to verify the transfection efficiency, CCK8 and cloning formation assay were used to detect cell proliferation and cloning ability, Transwell invasion and scratch assay were used to detect cell invasion and migration ability, and flow cytometry was used to explore cell cycle and apoptosis. Results Cell proliferation and cloning ability in the experimental group were significantly lower than those in the control group(P<0.01);cell invasion and migration ability in the experimental group were significantly lower than that in the control group(P<0.05);PDK1 gene silencing in SKOV3 cells shortened the cell replication periodand increased its early apoptotic ability(P<0.01). Conclusions PDK1 gene silencing significantly inhibits the proliferation, migration, invasion and cloning ability of SKOV3 cells, shorten the S phase of cells and increase the apoptotic rate.