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

作　　者：姚莎莎 尚文雯 徐睿 陈献 王芳[1] YAO Sha-sha;SHANG Wen-wen;XU Rui;CHEN Xian;WANG Fang(National Key Speciality of Clinical Laboratory,Department of Laboratory Medicine,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029;Department of Laboratory Medicine,the Affiliated Jiangning Hospital of Nanjing Medical University,Nanjing 211100,China)

机构地区：[1]南京医科大学第一附属医院检验学部国家临床检验重点专科,江苏南京210029 [2]南京医科大学附属江宁医院检验科,江苏南京211100

出　　处：《基础医学与临床》2019年第10期1381-1387,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81772779);江苏省六大高峰人才项目(2015-WSN-034);江苏省重点人才基金(ZDRCA2016003);江苏省“科教强卫工程”实验诊断学重点实验室(ZDXKB2016005)

摘　　要：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.

关 键 词：丙酮酸脱氢酶激酶1 卵巢癌 生物学行为 肿瘤代谢 

分 类 号：R33[医药卫生—人体生理学]