The Effect Of The Overexpression Of DLC-3 On The Growth,Migration And Apoptosis Of MCF-7 Cells

Objective: To explore the effect of DLC-3 over-expression on the cell biological behaviour such as growth,migration and apoptosis of human breast cancer cell MCF-7.To observe the potential role of DLC-3 in the process of breast cancer.Methods:(1)Human breast cancer cell MCF-7 was subcultured,which was 70-90% confluent at transfection with plasmid DNA.The cell was incubated for 48 h after transfection and the efficiency of the transfection was identified by inverted fluorescence microscope.The plasmid with DLC-3 was tansfected into experimental groups to up-regulate the expression of the gene.(2)Collect different groups of cell after transfecion.The m RNA expression and relative quantification of DLC-3 gene in different groups was detected by Real-time PCR.(3)The ability of cell proliferation and migration was observed after the transfection by using CCK-8 assay and wound healing in vitro;Flow cytometry was used to analysis the ratio of apoptotic cell in different groups.Results:(1)The efficiency of transfection is about 30-40% and Real-time PCR showed that the expression of DLC-3 was greatly increased compared with the control groups after transfection.(p<0.01).(2)CCK-8 assay showed that the over-expression of DLC-3 reduced the ability of cell proliferation in MCF-7.The absorptance of MCF-7 cell after plasmid with DLC-3 transfection was at a evidently lower level after cultured for 12 h,24h and 48 h compared with the control and negative control groups(p<0.01).(3)The healing area rate in experimental groups at 12 h was significantly less than the control groups(p<0.01)and negative control groups(p=0.09);the rate in experimental groups at 24 h was still significantly less than other groups(p<0.01).(4)Flow cytometry showed that the ratio of apoptotic cell in MCF-7 with DLC-3 expession was 15.893±1.597%,while the control group was 11.973±1.564% and the negative control group was 10.277±1.947.The ratio in experimental groups was significantly increased compared with other groups(p<0.05).Conclusion: In this study we showed direct evidence to the inhibition fuction of DLC-3 over-expression in the proliferation and migration process of MCF-7 cells.There was a higher ratio of apoptotic cell in the MCF-7 cells after up-regulated the expression of DLC-3 gene.