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目的:研究沉默MAD2基因对结肠癌细胞的影响。方法 :采用RT-PCR和Western印迹法筛选MAD2基因在7种结肠癌细胞中的高表达细胞株,并用siRNA瞬时转染以降低MAD2表达,随后采用CCK-8法检测结肠癌细胞增殖能力。流式细胞仪检测细胞凋亡和周期,transwell实验评估转染后结肠癌细胞迁移能力的改变。结果:SW1116细胞在mRNA和蛋白水平均呈现高表达MAD2,转染靶向MAD2的siRNA可显著降低MAD2在SW1116细胞中的表达。下调MAD2后,SW1116细胞的增殖能力较对照组有明显下降(P<0.05)。干扰siRNA组细胞凋亡比例较对照组增加,而细胞周期未见统计学差异。transwell实验中,siRNA干扰组穿膜细胞数为(45±2)个,显著低于阴性对照组[(185±5)个]和空白对照组[(195±5)个](P<0.01)。结论:SW1116高表达MAD2基因,沉默MAD2基因显著降低肠癌细胞增殖,诱导凋亡,并抑制细胞迁移。MAD2基因很可能成为新的预防结肠癌转移的目的基因。
Objective: To study the effect of silencing MAD2 gene on colon cancer cells. Methods: MAD2 gene was screened by RT-PCR and Western blotting in 7 colon cancer cell lines. Transient transfection with siRNA to reduce the expression of MAD2, and then using CCK-8 method to detect the proliferation of colon cancer cells. Flow cytometry was used to detect apoptosis and cycle. The transwell assay was used to evaluate the migration of colon cancer cells after transfection. Results: SW1116 cells showed high expression of MAD2 at mRNA and protein levels. Transfection of siRNA targeting MAD2 significantly reduced the expression of MAD2 in SW1116 cells. After MAD2 was down-regulated, the proliferation of SW1116 cells was significantly lower than that of the control group (P <0.05). The percentage of apoptosis in siRNA group was higher than that in control group, but there was no significant difference in cell cycle. Transwell experiments showed that the number of transmembrane cells in the siRNA interference group was (45 ± 2), significantly lower than that in the negative control group (185 ± 5) and the blank control group (195 ± 5) (P <0.01) . Conclusion: MAD2 gene is highly expressed in SW1116 and MAD2 gene silencing significantly reduces the proliferation, induces apoptosis and inhibits cell migration in colorectal cancer cells. MAD2 gene is likely to become a new target of prevention of colon cancer metastasis.