EDNRB基因真核表达载体的构建及其对MCF-7细胞增殖的影响

Construction of EDNRB Gene Eukaryotic Expression Vector and Its Effect on Proliferation of MCF-7 Cells

目的构建pCI2-EDNRB真核表达载体,探讨EDNRB过表达对MCF-7细胞增殖的影响。方法设计EDNRB基因特异性引物,应用PCR法扩增其编码序列,构建pCI2-EDNRB真核表达载体并转染MCF-7细胞,RT-PCR和Western blot分别检测EDNRB的mRNA和蛋白表达;MTT法检测EDNRB过表达后MCF-7细胞增殖能力。结果pCI2-EDNRB真核表达载体成功构建,MCF-7细胞中EDNRB的mRNA和蛋白水平均较对照组明显升高,过表达EDNRB抑制了MCF-7细胞的增殖。结论该研究成功构建了pCI2-EDNRB真核表达载体,过表达EDNRB可抑制MCF-7细胞增殖,这为深入研究EDNRB在乳腺癌中的功能及分子机制打下良好基础。

Objective To construct the eukaryotic expression vector pCI2-EDNRB and investigate the effect of EDNRB overexpression on the proliferation of MCF-7 cells.Methods EDNRB gene-specific primers were designed and amplified by PCR.The eukaryotic expression vector pCI2-EDNRB was constructed and transfected into MCF-7 cells.The mRNA and protein expression of EDNRB were detected by RT-PCR and Western blot,respectively.MTT assay The proliferation ability of MCF-7 cells after over-expression of EDNRB was examined.Results The eukaryotic expression vector of pCI2-EDNRB was successfully constructed.The mRNA and protein levels of EDNRB in MCF-7 cells were significantly higher than those in the control group.Overexpression of EDNRB inhibited the proliferation of MCF-7 cells.Conclusion The eukaryotic expression vector pCI2-EDNRB was successfully constructed in this study.Overexpression of EDNRB can inhibit the proliferation of MCF-7 cells,which lays a good foundation for further study of the function and molecular mechanism of EDNRB in breast cancer.