The Effects Of RNA Inteference To HER-2 Gene On Cell Cycle And Proliferation Of Human Breast Carcinoma Cells In Vitro

Objective:To investigate whether RNA interference(RNAi) induced by plasmid vector could suppress epidermal growth factor receptor 2(HER-2) expression in human breast carcinoma cells SKBr3 and the effect on cell cycle and proliferation.Methods:The small hairpin RNA(shRNA) eukaryotic expression vector targetingHER2gene,namedpshRNA-HER2-1/pshRNA-HER2-2,was constructed and transfected into cultured human breast carcinoma cell of line SKBr3 via lipofectamine2000;The inhibited effects on HER2 gene were determined by semi-quantitative reverse transcription PCR and flow cytometry.Cell cycle analysis was carried out via flow cytometry;The antiproliferative effects of pshRNA-HER2 were assessed by both MTT and colony assay.At the same time,the eukaryotic expression vector targeting GFP gene,named pshRNA-GFP was constructed as the positive control.Results: The successful construction of pshRNA-HER2-1/ pshRNA -HER2-2/ pshRNA-GFP plasmid was identified with sequencing.After the shRNA expression vector was successfully transfected into SKBr3 cells via lipofectamine2000,pshRNA-HER-1 reduced HER2 gene expression by 65.5%,.Compared with the control group, pshRNA-HER-1 reduced the cell numbers by 48.2% and decreased the colonies by 52.5% .Cell cycle analysis showed that pshRNA-HER2 induced accumulation of cells in...