SHP-1基因对人乳腺癌MDA-MB-231细胞增殖活性的影响

Inhibitory effect of SHP-1 gene transfer on the proliferation of breast cancer cell line MDA-MB-231

目的初步观察转染SHP-1基因前后MDA-MB-231细胞SHP-1蛋白表达情况及转染SHP-1基因对MDA-MB-231细胞增殖能力的影响。方法构建真核表达载体pEGFP-C3-SHP-1,利用脂质体转染法转染乳腺癌MDA-MB-231细胞,经G418筛选阳性克隆。用免疫细胞化学和Westernblot检测SHP-1基因在MDA-MB-231细胞中的稳定表达情况,并通过MTT法观察细胞生长曲线。结果蛋白水平证实,转染细胞内有SHP-1基因的高表达,转染后细胞增殖能力明显受抑制(P<0.05)。结论向无SHP-1基因表达的MDA-MB-231细胞内转染SHP-1基因并使其稳定表达,可以使MDA-MB-231细胞的增殖受到抑制,为以后进一步探讨SHP-1基因的功能打下了基础。

Objective To observe SHP-1 protein expression in breast cancer cell line MDA-MB-231 before and after SHP-1 gene transfer and its effect on the proliferation of MDA-MB-231 cells. Methods The eukaryotic expression vector pEGFP-C3-SHP-1 was constructed and transfected into breast cancer cell line MDA-MB-231 via LipofectamineTM2000, and the positive clones were selected using G418. SHP-1 expression in MDA-MB-231 cells was detected with immunocytochemistry and Western blotting, and the cell growth curve was observed using MTT assay. Results SHP-1 was highly expressed in transfected MDA-MB-231 cells, whose proliferation was significantly inhibited （P0.05）. Conclusion SHP-1 gene transfer into MDA-MB-231 cells results in inhibition of the cell proliferation.