PinX1基因真核表达载体的构建及其对乳腺癌MCF-7细胞增殖的抑制作用

探讨了PinX1 基因在乳腺癌MCF-7 细胞生长和细胞周期中的作用, 初步探讨了该基因用于乳腺癌临床治疗的可行性. 采用RT-PCR 技术从293-T 细胞中扩增PinX1 基因, 将其克隆入真核表达载体pEGFP-C1 中, 再将重组质粒转染MCF-7 细胞. 通过real-time PCR 检测PinX1 基因的mRNA 表达, 用MTT 法检测转染前后细胞生长曲线的变化, 用流式细胞仪检测转染目的基因后细胞生长周期的改变. 检测结果表明, PinX1 基因已经在转染后MCF-7 细胞的细胞核内稳定表达, 乳腺癌细胞生长明显减缓(P <0.05), 增殖变慢(P <0.05), 细胞生长阻滞于G0/G1 期, 说明PinX1 基因可抑制乳腺癌MCF-7 细胞的生长和增殖.

Construction of Recombinant Eukaryotic Expression Vector for PinX1 and Its Roles in Inhibiting Proliferation of Human Breast Cancer MCF-7 Cell Line

This paper explores the effects of PinX1 gene on the growth and the cell cycles of breast carcinoma cell line MCF-7 and the potential in treatment of breast cancer. The PinX1 mRNA was amplified from 293-T cell with RT-PCR and inserted into pEGFP-C1 vector. The recombined eukaryotic expression vector of PinX1 was confirmed by enzyme digestion, and transfected into the breast carcinoma cell line MCF-7 using a liposome method. The expressed mRNA was detected by real-time PCR, and the cell growth and cell cycles by MTT and flow cytometry, respectively. Results showed that PinX1 transfection into the breast cancer MCF-7 cell line was stable and the transfected gene was integrated into nucleolus DNA of transfected cells. Transfection of PinX1 gene could  significantly inhibit the growth and proliferation of breast cancer cells (P < 0.05) and the cell growth was arrested at G0/G1 stage. The conclusion is that PinX1 gene could inhibit growth and proliferation of breast cancer cells.