Effect Of DJ-1 On The Proliferation, Apoptosis And Invasion Of Ovarian Carcinoma SKOV3 Cells

Objective:The aim of this study was to explore the effect of DJ-1 on the proliferation,apoptosis and invasion of ovarian cancer SKOV3 cells after DJ-1 silencing by RNA interference.Methods:1. Design and synthesis three pairs of specific si RNA targeting DJ-1, and transiently transfected into SKOV3 cells via liposome transfection method. The study were divided into five groups:normal control group, negative control group,si RNADJ-1-a group,si RNA-DJ-1-b group,si RNA-DJ-1-c group.2. Western blotting was used to detect the expression of DJ-1 and Selecting the best interference sequence.3. The abilities of proliferation,apoptosis and invasion of DJ-1 were assessed by MTT assay,Flow cytometry and transwell assay, respectively.Results:1. Western blotting was used to detect the DJ-1 protein expression after transfection of si RNA. The results showed that the level of DJ-1 protein in the si RNA-DJ-1 groups were significantly lower than normal control group and negative control group(0.07±0.01、0.11±0.02、0.10±0.01 vs 0.51±0.02、0.50±0.02,P<0.05),while the relative DJ-1 protein expression levels of normal control group and negative control group showed no significant difference(0.51±0.02 vs 0.50±0.02,P>0.05).There was a significant difference of si RNA-DJ-1-a group compared to si RNA-DJ-1-b group and si RNA-DJ-1-c group(0.07±0.01 vs 0.11±0.02,0.07±0.01 vs0.10±0.01,P<0.05).2. MTT assay was used to detect cell proliferation. 24 h,48h and 72 h after transfection, the results showed that si RNA-DJ-1-a group cell proliferation rate was lower than normal control group(24h,0.379±0.044 vs 0.492±0.035;48h,0.486±0.059 vs 1.127±0.078;72h,0.738±0.046 vs 1.373±0.055, P<0.05) and negative control group(24h,0.379±0.044 vs 0.460±0.057;48h,0.486±0.059 vs 1.043±0.080;72h,0.738±0.046 vs 1.350±0.067, P<0.05).3. Flow cytometry was used to detect cell apoptosis. The results showed that the apoptosis rate of si RNA-DJ-1-a group was significantly higher than normal control group( 27.97±9.86% vs 1.77±0.71%, P<0.05) and negative control group(27.97±9.86% vs 2.83±0.55%, P<0.05), while the apoptosis rate of normal control group and negative control group showed no significant difference(1.77±0.71% vs2.83±0.55%,P>0.05).4. transwell assay was used to detect cell invasion. The results showed that the cell invasion force of si RNA-DJ-1-a group was significantly lower than normal control group( 27.67±1.86 vs 60.83±2.04, P<0.05) and negative control group(27.67±1.86 vs 58.83±1.47, P<0.05), while the cell invasion force of normal control group and negative control group showed no significant difference(60.83±2.04 vs58.83±1.47,P>0.05).Conclusion:Knocking down DJ-1 by si RNA could promoted cell apoptosis and inhibited cell proliferation and invasion.