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目的探讨抑制垂体瘤转化基因(PTTG)表达后胆管癌细胞体外生长的变化及其对5-FU化疗敏感性的影响。方法将反义PTTG真核表达载体pcDNA3.1-PTTGas和空白载体pcDNA3.1(+)转染入胆管癌细胞株QBC939中,筛选获得稳定表达株tQBC939(PTTG-)和tQBC939(pcDNA3.1),并设立未转染的QBC939细胞对照组,绘制细胞生长曲线,以MTT法检测细胞增殖力的变化,流式细胞仪检测细胞周期的比例变化,并采用5-FU处理细胞后,以MTT检测3组细胞的生长抑制率,计算IC50值,流式细胞仪和Hoechst染色法检测各组细胞凋亡情况。结果 tQBC939(PTTG-)组与tQBC939(pcD-NA3.1)组和QBC939组比较,前者生长较为迅速,S期细胞比例增多,G2/M期细胞比例减少(P<0.05),增殖指数较高(P<0.05);经5-FU处理后,tQBC939(PTTG-)的IC50值明显低于tQBC939(pcD-NA3.1)和QBC939细胞(P<0.05),细胞凋亡增多。结论转染PTTG反义DNA可增强胆管癌细胞的增殖力和对5-FU化疗的敏感性。
Objective To investigate the changes of in vitro growth of cholangiocarcinoma cells after inhibiting the expression of pituitary tumor transforming gene (PTTG) and its effect on the sensitivity of 5-FU chemotherapy. Methods Antisense PTTG eukaryotic expression vector pcDNA3.1-PTTGas and blank vector pcDNA3.1 (+) were transfected into cholangiocarcinoma cell line QBC939, and the stable expression strains tQBC939 (PTTG-) and tQBC939 (pcDNA3.1) , And set up untransfected QBC939 cells control group, plot the cell growth curve to detect cell proliferation by MTT assay changes in flow cytometry cell cycle, and 5-FU treatment of cells by MTT assay The inhibition rate of the growth of the three groups of cells was calculated. The IC50 value was calculated. The apoptosis of the cells in each group was detected by flow cytometry and Hoechst staining. Results Compared with tQBC939 (pcD-NA3.1) group and QBC939 group, the former developed rapidly, the proportion of S phase cells increased, the proportion of cells in G2 / M phase decreased (P <0.05), and the proliferation index was higher (P <0.05). After treatment with 5-FU, the IC50 of tQBC939 (PTTG-) was significantly lower than that of tQBC939 (pcD-NA3.1) and QBC939 cells (P <0.05). Conclusion Transfection of PTTG antisense DNA can enhance the proliferation of cholangiocarcinoma cells and its sensitivity to 5-FU chemotherapy.