下调fascin抑制卵巢癌细胞从G0/G1期向S期转变的实验研究

目的:研究下调聚束蛋白（fascin）对卵巢癌细胞周期、增殖和凋亡的影响。方法:卵巢癌细胞SKOV3感染阴性对照慢病毒载体、shRNA fascin 1慢病毒载体、shRNA fascin 2慢病毒载体，用Real-time PCR和Western blot检测干扰效果。MTT检测增殖活性，克隆形成实验检测细胞克隆形成能力，流式细胞术检测细胞周期和细胞凋亡，Western blot检测细胞中活化型Caspase-3（Cleaved Caspase-3）、细胞周期依赖性蛋白激酶4(cyclin-dependent kinase 4，Cdk4)、活化型Caspase-9（Cleaved Caspase-9）和p21蛋白水平。结果:shRNA fascin 1慢病毒载体、shRNA fascin 2慢病毒载体感染后的SKOV3细胞中fascin mRNA和蛋白表达水平均降低，并且shRNA fascin 1慢病毒载体感染后细胞中fascin mRNA和蛋白表达水平下降更多。下调fascin后的卵巢癌细胞增殖能力降低，细胞克隆形成能力也降低，细胞G0/G1期比例升高，细胞凋亡率升高，细胞中活化型Caspase-3、活化型Caspase-9蛋白水平升高，p21蛋白水平也升高，Cdk4蛋白水平降低，与感染阴性对照慢病毒载体和未经感染的细胞比较，差异有统计学意义（P<0.05）。结论:下调fascin可降低卵巢癌细胞增殖能力、抑制卵巢癌细胞从G0/G1期向S期转变并诱导卵巢癌细胞凋亡。

Down-regulation of fascin inhibits ovarian cancer cell transition from G0/G1 to S phase

Objective:To study the effect of down-regulation of fascin on cell cycle，proliferation and apoptosis of ovarian cancer.Methods:Ovarian cancer cells SKOV3 were infected with negative control lentiviral vector，shRNA fascin 1 lentiviral vector and shRNA fascin 2 lentiviral vector.The interference effects were detected by Real-time PCR and Western blot.MTT detected proliferation activity.Clone formation test was used to detect the ability of clone formation.Cell cycle and apoptosis were detected by flow cytometry.Western blot was used to detect the levels of Cleaved Caspase-3，Cleaved Caspase-9，cyclin-dependent kinase 4 (Cdk4) and p21 protein in cells.Results:Fascin mRNA and protein expression levels in SKOV3 cells infected with shRNA fascin 1 lentiviral vector and shRNA fascin 2 lentiviral vector decreased，moreover，the expression of fascin mRNA and protein decreased more after shRNA fascin 1 lentivirus vector infection.The proliferation ability of ovarian cancer cells was decreased after fascin down-regulation.The ability of cell cloning to form also was decreased.The proportion of G0/G1 phase was increased.The rate of apoptosis was increased.The level of Cleaved Caspase-3 and Cleaved Caspase-9 protein was increased in cells.The level of p21 protein also was increased.The level of Cdk4 protein was reduced.Compared with infected negative control lentivirus vector and uninfected cells，the difference was statistically significant (P<0.05).Conclusion:Down-regulation of fascin can reduce the proliferation of ovarian cancer cells，inhibit ovarian cancer cell transition from G0/G1 to S phase and induce apoptosis of ovarian cancer cells.