LRP16 Gene On Human Endometrial Carcinoma HEC-1-B Cells

Objective:To study the LRP16 gene carrying the eukaryotic expression plasmid EX-Y2069-M29,by cationic liposome transfection HEC-1-B cells in transfection efficiency, to understand before and after transfection the expression of LRP16 gene,to observe LRP16 on the HEC-1-B cell proliferation. Methods:The eukaryotic expression vector LRP16 EX-Y2069-M29 and EX-NEG-M29 empty plasmids were transfected by liposome cultured HEC-1-B cells transfected with empty vector EX-NEG-M29's HEC-1-B as an empty vector control, to HEC-1-B cells as negative control, was observed under a fluorescence microscope, fluorescent effect of transfection using RT-PCR detected before and after transfection LRP16 gene expression, depicted before and after transfection cells growth curve, with the WST-1 method to detect cell proliferation. Results:1.LRP16 group, empty vector group, some cells in the hair of green fluorescence under fluorescence microscope, and the negative control group, no green fluorescent cells.2. Blank control group, empty vector group and the transfected HEC-1-B cells LRP16mRNA the relative expression level was 0.1231±0.0366,0.1486±0.0287 and 0.5060±0.0198, plasmid transfection group compared with the other two groups,the difference was statistically significant, (F= 161.547, P= 0.0003).3. Observe HEC-1-B cell growth curve, not transfected HEC-1-B cell population doubling time of 0.8 days logarithmic phase in between 2-6 days. Transfected with LRP16 the HEC-1-B cell population doubling time of 2 days, the logarithmic growth phase in between 4-6 days, significantly lower than non-transfected cells.4.LRP16 group, empty vector group and negative control HEC-1-B cells WES-T colorimetry, LRP16 transfected HEC-1-B cells continued to proliferate in vitro, but with the empty vector group and negative control compared, HEC-1-B cell proliferation is decreased; empty vector group and negative control group HEC-1-B cell proliferation capacity was not changed.Conclusion:1. EX-Y2069-M29 plasmid can be successfully transferred by liposome HEC-1-B cells, and access to effective and stable expression; 2. Transfected HEC-1-B cells LRP16 gene expression was significantly increased; 3. Transfected HEC-1-B cell growth rate decreased.