【摘要】 目的探讨S-腺苷甲硫氨酸脱羧酶(SAMDC)基因沉默对乳腺癌MCF-7细胞增殖及侵袭能力的抑制作用。方法用特异性沉默SAMDC基因表达的RNA干扰真核表达载体SAMDCshRNA表达载体(pGPU6-SAMDC)转染乳腺癌MCF-7细胞,采用RT-PCR和Western-blot技术分别检测SAMDC mRNA和蛋白质表达水平;细胞增殖实验(CCK-8法)和细胞周期分析评价SAMDC基因沉默对乳腺癌MCF-7细胞增殖的影响,肿瘤侵袭实验观察SAMDC基因沉默对乳腺癌细胞MCF-7侵袭活性的影响。结果SAMDCsh RNA表达载体转染后,MCF-7细胞SAMDC mRNA水平下调43.8%,蛋白表达水平下降66.5%;MCF-7细胞的增殖活性和侵袭能力均受到明显抑制,细胞增殖活性下降37%,细胞周期分析G1期细胞增加。结论SAMDC基因沉默可通过延长细胞周期的G1期而抑制乳腺癌MCF-7细胞的增殖,并能降低癌细胞侵袭活性,提示SAMDC可作为乳腺癌基因治疗的靶分子。更多还原

【Abstract】 Objective To investigate the inhibitory effect of the S-adenosylmethionine decarboylase(SAMDC)gene silencing on proliferation and invasion of breast cancer MCF-7 cells.Methods Breast cancer MCF-7 cells were transfected with a short hairpin RNA eukaryotic expression vector against SAMDC(pGPU6-SAMDC).The effect of suppressing proliferation of MCF-7 cells was detected by cell proliferation assay(CCK-8)and cell cycle analysis.Semi-quantitative RT-PCR analysis of mRNA and western blot analysis of proteins were used to determine expression of the SAMDC gene.The invasive ability of breast cancer MCF-7 cells was assessed by invasion assay in vitro.Results After the shRNA expression vector was transfected into breast cancer MCF-7 cells,the growth and invasive ability of MCF-7cells were significantly inhibited,and the proliferation of cells was decreased by 37%.Gene expression of SAMDC in MCF-7 cells was also significantly inhibited,of mRNA was inhibited by 43.8% and of protein was inhibited by 66.5%.Moreover,the proportion of MCF-7 cells in the G0/G1 phase was increased.Conclusion The shRNA of the SAMDC gene can effectively inhibit the proliferation and invasion of MCF-7 cells and is a potential target in gene therapy of breast cancer.更多还原