青藤碱干预肿瘤细胞凋亡的erkbcl2信号转导机制研究-erk bcl 2 signal transduction mechanism of sinomenine intervening tumor cell apoptosis.docxVIP

中文摘要 目的：研究青藤碱干预肿瘤细胞凋亡的信号转导机制。观测不 同剂量青藤碱干预肿瘤细胞凋亡的效果。方法：体外培养人宫颈癌 Hela 细胞，分高、中、低 3 个剂量组，各剂量组加入含不同浓度青 藤碱的 RPMI1640 培养液，空白对照组加入不含青藤碱的 RPMI1640 培养液。培养 72 小时后，提取细胞总蛋白，用 Western blot 方法 检测细胞 COX-2、p-ERK、Bcl-2 蛋白表达情况。初步研究探讨青藤 碱通过何种信号转导通路干预肿瘤细胞增殖。给药后观察细胞一般 情况，用 MTT 方法检测细胞生长情况，了解青藤碱对人宫颈癌 Hela 细胞增殖的影响。用流式细胞仪检测细胞凋亡情况。结果表明：1. 中、高剂量实验组细胞生长减缓。与空白组比较，中、高剂量实验 组肿瘤细胞 MTT 的 OD 值减小（P＜0.05）；2.中高剂量组 COX-2 蛋白 表达下降。与空白组比较，中、高实验组 COX-2 蛋白表达下降，差 别具有统计学意义（P＜0.05）；3. 高剂量实验组 p-ERK 蛋白表达下 降。与空白组比较，差别具有统计学意义（P＜0.05）；4. 中高剂量 实验组 Bcl-2 蛋白表达下降。与空白组比较，差别具有统计学意义（P＜0.05）。5.与空白组比较，实验组细胞凋亡增加（P＜0.05）， 说明青藤碱作用后，细胞凋亡率升高。结论：1.青藤碱可抑制 Hela 细胞增殖。 2.青藤碱可诱导 Hela 细胞凋亡，可能是通过减少 Bcl-2 蛋白表达实现这个作用。3.青藤碱可能抑制 COX-2、ERK-Bcl-2 信号 转导通路的功能。 关键词：青藤碱 Hela 细胞 COX-2 Bcl-2 p-ERK 凋亡 ABSTRACTObjective: To research SIN intervention the signal transduction of tumour cell. Study different dose SIN induce apoptosis of tumour cell. Methods: Hela cell were incubeded in vitro.This cell were divided into 3 experimental groups and 1 control group. The experimental group were given RPMI1640 with different dose SIN. The control group were given RPMI1640 without SIN. After incubated 72 hours, The total protein ofHela cell was extracted. Detection COX-2、p-ERK、Bcl-2 by Western blot.To approach which signal transduction were affect in SIN interfere cell proliferation. To observe the general state of Hela cell and the Flow Cytometer were used to detect apoptosis of Hela cell. Results: 1. The grow speed of middle and high dose experimental groups was limted. Thevalue OD of these groups were lower than this of control group（P＜0.05);2. The expression of COX-2 in middle and high dose experimental groups were decrease, compare with control group(P ＜ 0.05);3.Theexpression of p-ERK in high dose experimental groups was decrease, compare with control group(P＜0.05).But not decrease in low dose group. 4.The expression of Bcl-2 in middle and high dose experimental groups were decrease, compare with control group(P＜0.05) 5. The apoptosis rate we