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机构地区:[1]牡丹江医学院生化教研室,黑龙江牡丹江157011
出 处:《牡丹江医学院学报》2007年第6期12-14,共3页Journal of Mudanjiang Medical University
基 金:黑龙江省卫生厅指导项目(2006-400);牡丹江医学院指导项目(C2006-09)
摘 要:目的:从基因水平探讨白花蛇舌草多糖对体外培养的人肝癌Bel7402细胞凋亡诱导的作用及机制。方法:水提醇沉法提取白花蛇舌草多糖,人肝癌Bel7402细胞常规体外细胞培养,设定空白对照组、5-氟尿嘧啶阳性对照组、白花蛇舌草多糖组。HE染色法光镜下观察细胞形态改变,计算凋亡指数,采用MTT法检测白花蛇舌草多糖对癌细胞的生长抑制作用,RT-PCR法检测原癌基因bc l-xl、抑癌基因p53基因mRNA表达的变化。结果:白花蛇舌草多糖体外抑制人肝癌Bel7402细胞生长、促进细胞凋亡,凋亡指数达16.97(,以MTT显色法检测细胞抑制率达到61.5(,以RT-PCR半定量法检测,上调p53基因mRNA表达,降低bc l-xl基因mRNA表达,以上各指标与空白对照组相比均有显著差异。结论:白花蛇舌草多糖抑制人肝癌Bel7402细胞增长,诱导细胞凋亡,其分子机制可能与激活抑癌基因p53基因、抑制原癌基因bc l-xl基因表达有关。Objective:To study the inducing apoptosis of Hedyotis diffusa extract (HDP) on Human Hepatocellular carcinoma cell line Bel7402 and the molecular mechanism. Methods: Human Hepatocellular carcinoma cell line Bel7402 was used in vitro cell culture,there were 3 groups:control group, HDP group and 5-FU group, the Bel7402 cells were dealed with them respectively. The apoptosis morphologic changes of Human Hepatoeellular carcinoma Cells were observed by Invert microscope, the apoptosis rate were detected with hematoxylin stain by light microscope.. Inhibition of Be17402 cell proliferation was measured with MTF assay. The expression of oncogene bcl - xl and anti - oncogene p53 of Be17402 cells was observed with RT - PCR assay. Results: Compared with the control group, the condensation of nuclear,vacuolar degeneration of mitochondria could be found through light microscope. The apopto- sis rate of HDP group was remarkably increased with it in control group (p 〈0.01 ), The Inhibition of HDP group was remarkably increase with it in control group ( p 〈 0.01 ). The expression of the gene p53 raised and bcl - xl decreased by RT-PCR. Conclusion.HDP is able to lead to tumor cels apoptosis,the activation of p53 and the suppression of bcl-xl may contribute to the apoptosis mechanism.
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