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作 者:刘永年[1] 王索安[2] 赵明[1] 张伟[1] 赵海龙[1] 李菊英[1]
机构地区:[1]青海大学医学院 [2]浙江医学高等专科学校病理教研室
出 处:《青海医学院学报》2008年第1期1-6,共6页Journal of Qinghai Medical College
摘 要:目的研究氟尿嘧啶(5-Fluorouracial,5-FU)体外影响人大肠癌Lovo细胞增殖抑制及诱导凋亡的分子机制。方法采用四甲基偶氮唑盐(MTT)显色法,检测不同浓度5-FU作用不同时间对Lovo细胞生长所产生的不同影响;光镜观察凋亡细胞的形态特点;流式细胞术(FCM)检测凋亡细胞百分率;琼脂凝胶电泳、免疫组化SP法检测Lovo细胞增殖核抗原Ki-67及凋亡相关基因蛋白p53、Fas表达水平。结果5-FU能抑制Lovo细胞生长,在一定剂量和时间范围内,通过光镜观察,见凋亡细胞明显增多,经5-FU诱导后,用FCM分析Lovo细胞,发现凋亡百分率增加并显示剂量和时间效应。实验组细胞Ki-67表达明显低于对照组,Fas表达明显升高,而P53在诱导前后均无表达。结论5-FU能诱导Lovo细胞凋亡并抑制其增殖及侵袭性,Fas基因激活可能是触发Lovo细胞凋亡的主要机制。Objective To study the molecular mechanism for 5-Fluorouracial(5-FU) and clarify whether it inhibit proliferation and induce apoptosis of colon cancer Lovo cells.Methods MTT Chromatography is adopted to test effects of 5-FU of different concentration and at different time of reaction over growth of Lovo cells,photomechanical method used to observe configuration and characteristics of apoptosis cells,FCM employed to test the percentage of apoptosis cells,electrophoresis and immunized grouping SP applied to test antigen Ki-67 of Lovo cells proliferation and expression of P53 and Fas genetic proteins in relation to apoptosis.Results 5-FU inhibits growth of Lovo cells,it is found that within a limited dosage and period of time,there has been an obvious increase in apoptosis cells under the photomechanical scope,the analysis of Lovo cells by FCM showed that after the induction of 5-FU,the percentage of apoptosis cells has increased,and FCM also displays dosage and time effects.Electrophoresis was obviously in a stripe shape of trapeziform.In the project group,Ki-67 expression in cells is clearly lower than that in the control group,and Fas expression in cells apparently tends to rise,whereas P53 has no expression before and after the induction.Conclusion 5-FU can induce apoptosis of Lovo cells and inhibit its proliferation and attack,activation of Fas gene might be the primary mechanism of triggering Lovo cells to apoptosis.
关 键 词:氟脲嘧啶 大肠癌 细胞凋亡 流式细胞术 琼脂凝胶电泳
分 类 号:R329.28[医药卫生—人体解剖和组织胚胎学] R735.34[医药卫生—基础医学]
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