Influence of clusterin gene silencing on proliferation and invasion of human ovarian carcinoma SKOV3 cell line / 第二军医大学学报

ABSTRACT

Objective: To investigate the influence of secretory clusterin(sCLU) gene silencing by RNA interference on the proliferation, apoptosis and invasion of human ovarian carcinoma SKOV3 cell line. Methods: The human ovarian carcinoma SKOV3 cells were transfected with sCLU-targeting siRNA by Lipofectamine 2000. The study included four groups the experimental group (sCLU-siRNA+Lipofectamine 2000), negative group I (negative control siRNA+Lipofectamine 2000), negative group II (sCLU-siRNA) and blank control group. The expressions of mRNA and protein were examined by RT-PCR and Western blotting analysis. The proliferation, apoptosis and invasion ability of SKOV3 cells were evaluated by MTT assay, flow cytometry and Transwell assay. Results: The sequence-specific siRNA of sCLU effectively and specifically down-regulated the expression of sCLU at both mRNA and protein levels. The MTT results showed that the proliferation of cells in the experimental group was significantly inhibited compared with those in the other groups (P < 0.05). Flow cytometry demonstrated that the apoptotic rate of cells in the experimental group([15.84±1.53]%) increased by 9% compared with those in the rest groups (P < 0.01). We also found that the matrigel penetrating capacity of cells in the experimental group ([26.52±6.22] cells/field) was significantly inhibited compared with those in the rest groups(P < 0.01). Conclusion: Silencing of sCLU can effectively inhibit the proliferation and invasion of human ovarian carcinoma SKOV3 cells, and it can also greatly promote their apoptosis, making sCLU gene a potential new target for ovarian carcinoma treatment.