miR-20b对卵巢癌细胞迁移和侵袭的影响及其机制

Effects of miR-20b on migration and invasion in ovarian carcinoma cells

目的:研究miR-20b在卵巢癌细胞系中的表达及其对迁移和侵袭的影响。方法:采用qRT-PCR检测卵巢癌细胞系HO8910,A2780,SKOV3与正常卵巢细胞系Hose中miR-20b的表达水平。将A2780细胞系分成两组,miR-20b模拟物组和阴性对照组,分别转染miR-20b mimics和阴性对照质粒;细胞划痕实验和Transwel l实验分别测定两组细胞迁移和侵袭能力,qRT-PCR和Western印迹分别测定两组血管内皮细胞生长因子(vascular endothelial cell growth factor,VEGF)mRNA和蛋白表达水平。结果:miR-20b在卵巢癌细胞系HO8910,A2780,SKOV3中低表达,分别为0.30±0.05,0.23±0.03及0.10±0.02,显著低于Hose细胞系的1.00±0.03(P<0.001)。miR-20b模拟物组划痕愈合率为23.20%±3.50%,阴性对照组为65.70%±8.30%,miR-20b模拟物组划痕愈合率显著低于阴性对照组(P<0.01)。miR-20b模拟物组侵袭细胞数为(21.3±4.5)个,阴性对照组为(73.5±6.7)个,miR-20b模拟物组侵袭细胞数显著少于阴性对照组(P<0.01)。miR-20b模拟物组VEGF mRNA相对表达量为0.30±0.02,而阴性对照组为1.00±0.05,miR-20b模拟物组VEGF mRNA表达量显著低于阴性对照组(P<0.001)。miR-20b模拟物组VEGF蛋白表达量为118.00±8.00,显著低于阴性对照组的180.00±5.90(P<0.05)。结论:miR-20b在卵巢癌细胞系中低表达,过表达miR-20b抑制卵巢癌细胞转移与侵袭,可能通过下调VEGF发挥抑癌作用。

Objective： To analyze the expression of miR-20b in ovarian carcinoma cell line and its effect on migration and invasion of ovarian carcinoma cell line. Methods： The expressions level of miR-20b in HO8910, A2780, SKOV3 and Hose was measured by qRT-PCR. The A2780 cell line was divided into two groups, miR-20b mimic group transfect with miR-20b mimics and negative control group transfect with miR-20b scramble by Lipofectamine 2000. The cell wound scratch assay was used to detect the ovarian cell migration ability. The cell invasion ability was detected by Transwell assay. The expression level of vascular endothelial cell growth factor （VEGF） mRNA and protein was measured by qRT-PCR and Western blot, respectively. Results： The expression of miR-20b in three ovarian carcinoma cell lines HO8910, A2780 and SKOV3 was significantly lower than normal cell line, Hose （0.30±0.05, 0.23±0.03, 0.10±0.02 vs 1.00±0.03）, the difference was statistically significant （P〈0.001）. The wound healing rate of miR-20b mimics group was 23.20%±3.50%, while 65.70%±8.30% in negative control group, the wound healing rate was significantly lower than negative control group （P〈0.001）. The invasive cell number in miR-20b mimics group was 21.30±4.50, while 73.50±6.70 in negative control group, the invasive cell number in miR-20b mimics group was significantly less than that in negative control group （P〈0.01）. The expression level of VEGF mRNA in miR-20b mimics group was 0.30±0.02, which was significantly less than 1.00±0.05 in negative control group （P〈0.001）. The expression level of VEGF protein in miR-20b mimics group was 118.00±8.00, which was significantly less than 180.00±5.90 in negative control group （P〈0.05）. Conclusion： MiR-20b has a lower expression in ovarian carcinoma cell lines. MiR-20b inhibits migration and invasion of ovarian carcinoma cells, which may through inhibiting the expression of VEGF.