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—timePCR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.
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.