【摘要】 目的探究白花蛇舌草对宫颈癌细胞增殖、端粒酶活性及Ki-67基因表达的影响。方法分别以10%,20%,40%白花蛇舌草含药血清处理人宫颈癌HeLa细胞并分别作为低、中、高剂量实验组,对照组细胞则以等量生理盐水处理,各组细胞干预后分别继续培养24,48,72 h。采用MTT法检测各组宫颈癌HeLa细胞增殖情况,采用TUNEL法检测各组宫颈癌HeLa细胞凋亡情况,采用流式细胞仪法检测HeLa细胞端粒酶活性,采用RT-PCR法检测宫颈癌HeLa细胞中Ki-67 mRNA表达情况。结果各实验组宫颈癌HeLa细胞增殖抑制率及凋亡率均随白花蛇舌草含药血清作用浓度增大及培养时间的延长呈逐渐升高趋势(P均<0.05);培养48 h后,对照组及低、中、高剂量实验组宫颈癌HeLa细胞端粒酶表达率分别为(30.41±1.96)%、(22.02±1.56)%、(11.31±1.42)%、(7.23±1.06)%,各实验组宫颈癌HeLa细胞端粒酶表达率均显著低于对照组(P均<0.05),且随白花蛇舌草含药血清作用浓度增大呈逐渐降低趋势(P<0.05)。与对照组比较,各实验组培养不同时间宫颈癌HeLa细胞中Ki-67 mRNA相对表达量均显著降低(P均<0.05),且均随白花蛇舌草含药血清作用浓度增大及培养时间的延长呈逐渐降低趋势(P均<0.05)。结论白花蛇舌草含药血清可显著抑制宫颈癌HeLa细胞增殖,诱导其凋亡,且具有显著的时间-剂量依赖性,其作用机制可能与抑制宫颈癌HeLa细胞端粒酶活性及显著下调Ki-67基因表达相关。更多还原

【Abstract】 Objective It is to investigate the effects of Hedyotis diffusa on the proliferation, telomerase activity and Ki-67 gene expression in cervical cancer cells. Methods Human cervical cancer HeLa cells respectively treated with 10%, 20% and 40% Hedyotis diffusa drug-containing serum were as low, medium and high dose of experimental group respectively. The cells were treated with the same amount of normal saline in the control group. All the cells were continuously cultured for 24, 48, and 72 hours after the treatment. The proliferation of HeLa cells of cervical cancer was detected by MTT assay. The apoptosis of HeLa cells of cervical cancer was detected by TUNEL method. The telomerase activity of HeLa cells was detected by flow cytometry. The expression of Ki-67 mRNA in HeLa cells of cervical cancer was detected by RT-PCR. Results The proliferation inhibition rate and apoptosis rate of cervical cancer HeLa cells in each experimental group increased gradually with the increase of the concentration of the serum containing Hedyotis diffusa and the prolongation of culture time(P<0.05). After 48 hours of culture, the telomerase expression rates of HeLa cells in the control group and the low, medium and high dose of experimental groups were(30.41±1.96)%,(22.02±1.56)%,(11.31±1.42)%, and(7.23±1.06)%, respectively. The telomerase expression rate of cervical cancer HeLa cells in each experimental group was significantly lower than that in the control group(P<0.05), and it gradually decreased with the increase of concentration of Herba Hedyotis herb serum(P<0.05). Compared with the control group, the relative expression of Ki-67 mRNA in cervical cancer HeLa cells was significantly decreased in each experimental group(P<0.05), and the expression gradually decreased with the increase of concentration of the drug-containing serum of Hedyotis diffusa and culture time(P<0.05). Conclusion The serum containing Hedyotis diffusa can significantly inhibit the proliferation and induce apoptosis of cervical cancer HeLa cells with significant time-dose-dependence. The mechanism may be related to inhibiting telomerase activity and significantly down-regulating Ki-67 gene expression in cervical cancer HeLa cells.更多还原