bcl-2+sirna对结肠癌细胞ht-29凋亡的作用机制研究 - 中华医学会.docVIP

bcl-2+siRNA对结肠癌细胞HT-29凋亡的作用机制研究 邱 进 河南省濮阳市油田总医院 普外二科 河南省濮阳市 457001 【作者信息】邱 进（1980.11.20），山东省阳谷县，男，本科，主治医师，主要研究方向是胃肠疾病的诊治（普外—胃肠外科） 【通讯作者】河南省濮阳市大庆路124号 河南省濮阳市油田总医院普外二科 457001 【摘要】目的：研究bcl-2+siRNA对结肠癌细胞HT-29的凋亡作用机制。方法：把HT-29细胞分成三组：实验组、空白对照组和阴性对照组。然后对实验组进行bcl-2+siRNA转染处理，对空白对照组不进行任何处理，对阴性对照组采用阴性对照siRNA转染处理。然后对三组细胞分别采用MTT比色法、流式细胞术、Western blot和分光光度法检测HT-29细胞的活力、细胞凋亡、Bcl-2和Bax蛋白的表达以及天冬氨酸和半胱氨酸蛋白酶中Caspase3和Caspase9的活化程度，把三组数据进行比照分析。结果：bcl-2+siRNA实验组细胞死亡率为57%，Caspase3和Caspase9的活化程度分别为98%和97%，远远高于空白对照组和阴性对照组。实验组Bcl-2和Bax蛋白的表达和细胞活力明显低于其它两组，P0.05具有统计学意义。但是空白对照组和阴性对照组比较差异不大。结论：bcl-2+siRNA通过促进促凋亡蛋白的表达和增强Caspase3、Caspase9的活化程度从而促使癌细胞凋亡，效果显著，可对结肠癌患者大规模推广。 【关键词】bcl-2+siRNA； 结肠癌HT-29细胞； 作用机制 Mechanism of bcl-2 + siRNA on HT-29 apoptosis in colon cancer cells [Abstract]Objective: To discuss the mechanism of bcl-2 + siRNA on HT-29 apoptosis in colon cancer cells.Method: The HT-29 human colon cancer cells were divided into three groups: experimental group,blank control group and negative control group.The experimental group was given bcl-2 + siRNA transfection process,the blank control group was not given treatment,the negative control group was given negative control siRNA transfection process.Three groups of cells were evaluated by MTT assay, flow cytometry, Western blot and spectrophotometry to detect HT-29 cell viability, apoptosis, Bcl-2 and Bax protein expression and activation of Caspase9 and Caspase3 in aspartic acid and cysteine proteases,and the three sets of data were analyzed.Results: Mortality rate of bcl-2 + siRNA experimental group was 57%,activation of Caspase3 and Caspase9 were 98% and 97%,and they were much higher than the blank control group and negative control group.Bcl-2 and Bax protein expression and cell viability of experimental group were significantly lower than the other two groups(P0.05), they were statistically significant.But differences between blank control group and negative control group were not significant. Conclusion:bcl-2 + siRNA promote expre