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目的研究miR-101对结肠癌细胞增殖能力的影响及与分子学机制。方法人工干预miR-101在结肠癌细胞株HCT116中的表达,分别转染阴性对照物(对照组)、miR-101抑制物(实验Ⅰ组)、miR-101模拟物(实验Ⅱ组)48 h后,用细胞增殖法分析其对人结肠癌HCT116细胞增殖能力的影响。用免疫印迹法测定miR-101在调节Notch1表达中的作用。结果实验Ⅰ组、实验Ⅱ组及对照组的miR-101表达量分别为0.467,1.767,0.967。与对照组相比,mimic-miR-101使HCT116细胞中miR-101的表达水平明显提高,差异有统计学意义(P<0.05)。实验Ⅱ组细胞miR-101的表达水平较对照组显著降低,差异有统计学意义(P<0.05)。在72 h,实验Ⅰ组、实验Ⅱ组及对照组OD值分别为1.10,0.76,0.91。在96 h,实验Ⅰ组、实验Ⅱ组及对照组OD值分别为1.57,0.92,1.20。与对照组相比,实验Ⅱ组的细胞增殖能力明显被抑制,差异均有统计学意义(均P<0.05)。miR-101调节结肠癌细胞HCT116增殖的影响是通过靶向调节Notch1的表达来实现的。结论在结肠癌发生及进展过程中,miR-101发挥抑癌基因作用,可靶向干扰Notch1的表达,进而负性调节结肠癌细胞的增殖。
Objective To study the effect of miR-101 on the proliferation of colon cancer cells and its molecular mechanism. Methods The expression of miR-101 in human colon cancer cell line HCT116 was induced by artificial intervention and transfected into the negative control (control group), miR-101 inhibitor (experimental group I) and miR-101 mimics (experimental group II) After that, the cell proliferation assay was used to analyze the effect on proliferation of human colon cancer HCT116 cells. The role of miR-101 in the regulation of Notch1 expression was determined by Western blotting. Results The expression of miR-101 in experimental group I, experimental group II and control group were 0.467, 1.767 and 0.967, respectively. Compared with the control group, mimic-miR-101 significantly increased the expression level of miR-101 in HCT116 cells, with statistical significance (P <0.05). The expression level of miR-101 in group Ⅱ was significantly lower than that in control group (P <0.05). At 72 h, the OD of experimental group I, experimental group II and control group were 1.10, 0.76, 0.91 respectively. At 96 h, the OD values of experimental group I, experimental group II and control group were 1.57, 0.92 and 1.20 respectively. Compared with the control group, the cell proliferation ability of group Ⅱ was significantly inhibited (all P <0.05). The effect of miR-101 in regulating the proliferation of colon cancer cell HCT116 is achieved by targeting the expression of Notch1. Conclusion In the process of carcinogenesis and progression of colon cancer, miR-101 plays a role of tumor suppressor gene and can specifically interfere with the expression of Notch1, thereby negatively regulating the proliferation of colon cancer cells.