重组质粒pDsRed1-C1-PinX1的建立及其在乳腺癌MCF-7细胞中的表达

Construction of recombinant plasmid pDsRed1-C1-PinX1 and its expression in breast carcinoma MCF-7 cells

目的构建PinX1真核表达载体,观察PinX1基因对乳腺癌MCF-7细胞生长和增殖的影响。方法重组质粒转染MCF-7细胞并筛选稳定表达系;用Real-timePCR检测PinX1基因mRNA的表达;MTT法检测转染前后细胞生长变化;平板克隆形成试验检测PinX1对MCF-7集落形成能力的影响。结果构建了真核表达载体PDsRed1-C1-PinX1的稳定表达系;转染后乳腺癌MCF-7细胞生长明显减缓,集落形成能力降低。结论 PinX1基因可抑制乳腺癌MCF-7细胞的生长和增殖。

Objective To observe the effect of PinX1 gene on the growth and proliferation of MCF-7 breast carcinoma cell line by constructing the eukaryotic expression vector ofPinX1. Methods MCF-7 ceils were transfected by recombinant plasmid and stable expression cells were then screened. PinX1 mRNA expression, growth and colony forming ability of MCF-7 cells were detected by Real-time PCR, MTT and plate clone formation assays, respectively. Results The constructed eukaryotic vector pDsRedl-C1-PinX1 expressed PinX1 stably. The growth and colony-forming ability of MCF-7 cells decreased after the transfection ofPinX1 gene. Conclusion PinX1 can inhibit the growth and proliferation of MCF-7 cells.