下调HEATR3表达对结直肠癌HCT116细胞增殖及Akt信号通路的影响

摘要

目的 观察下调HEATR3表达对结直肠癌HCT116细胞增殖以及Akt信号通路的影响,探讨HEATR3在结直肠癌中的作用.方法 将HCT116细胞随机分为观察组和对照组,分别转染HEATR3的干扰RNA及无义序列;采用MTT法检测细胞增殖的光密度(OD)值,采用实时荧光定量PCR法检测HEATR3 mRNA,Western blot法检测HEATR3、Akt、p-Akt.结果 两组转染前细胞增殖无差异,与对照组同时点比较,观察组转染后24、36、48 h时细胞增殖的OD值均降低(P均<0.05);转染48 h,观察组HEATR3 mRNA及蛋白表达量分别为0.49±0.04、0.03±0.01,对照组分别为2.08±0.45、0.47±0.01,P均<0.05.观察组Akt、p-Akt蛋白表达量分别为0.49±0.01、0.05±0.01,对照组分别为0.96±0.01、0.92±0.01,P均<0.05;结论 下调HEATR3表达可通过抑制Akt信号通路降低结直肠癌细胞的增殖能力,HEATR3可能是结肠癌发生、发展中的一个重要癌基因.%Objective To observe the effects of down-regulating HEATR3 on cell proliferation and Akt signaling pathway in colorectal cancer cell line HCT116, and to determine the role of HEATR3 in the colorectal cancer.Methods HCT116 cells were randomly divided into the observation group and control group which were separately transfected with the siRNA of HEATR3 and nonsense sequence.The optical density ( OD) value of cell proliferation was detected by MTT as-say, the expression level of HEATR3 mRNA was detected by real-time fluorescent quantificative PCR, and the protein lev-els of HEATR3, Akt and p-Akt were detected by Western blotting.Results There was no difference in the cell prolifera-tion between the two group before transfection.The OD values of cell proliferation in the observation group were lower than those of the control group at 24 h, 36 h and 48 h after transfection (all P<0.05);the HEATR3 mRNA and protein levels in the observation group were 0.49 ±0.04, 0.03 ±0.01, and those were 2.08 ±0.45 and 0.47 ±0.01 in the control group at 48 h after transfection (all P<0.05).Akt and p-Akt protein levels in the observation group were 0.49 ±0.01 and 0.05 ±0.01, and those were 0.96 ±0.01 and 0.92 ±0.01 in the control group after transfection (all P<0.05). Conclusion The down-regulation of HEATR3 can decrease the cell proliferation of colorectal cancer cells through inhibi-ting Akt signaling pathway, and HEATR3 may act as an important oncogene in the occurrence and development of colorectal cancer.