雷丸蛋白pPeOp诱导胃癌细胞SGC-7901凋亡机制研究  

Apoptosis mechanism of gastric cancer cell SGC-7901 induced by Omphalia lapidescens protein pPeOp

作　　者：赵肖涯 陆仲夏 杜丽君[1] 梁浩威 陈宜涛[1] 

机构地区：[1]浙江中医药大学生命科学学院,浙江杭州310053

出　　处：《中国药理学通报》2017年第9期1271-1277,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81374023);浙江省自然科学基金资助项目(No Y207765);浙江省大学生科技创新活动计划暨新苗人才计划项目(No 2016R410039)

摘　　要：目的探究雷丸蛋白pPeOp诱导人胃癌细胞SGC-7901的凋亡机制。方法采用CCK-8法和流式细胞术检测不同浓度的pPeOp(30、60、90 mg·L^(-1))对人胃癌细胞SGC-7901的活性抑制和凋亡诱导作用;q RT-PCR和Western blot检测死亡受体通路和线粒体通路TNF-R1、Fas/Fas L、Bcl-2、caspase-3、caspase-8的m RNA和蛋白表达情况。结果CCK-8结果显示,与Control组相比,阴性对照聚乙烯吡咯烷酮(PVP)组细胞存活率差异无显著性;阳性对照5-氟尿嘧啶(5-Fu)组细胞存活率为(53.71±7.34)%,存活率明显下降(P<0.05);30、60、90 mg·L^(-1)的pPeOp组细胞存活率分别为(80.95±6.25)%、(53.48±5.70)%、(44.61±6.50)%,存活率降低且与浓度呈负相关(r=0.984,P=0.016)。流式细胞术结果显示,PVP组与Control组相比差异无显著性,5-Fu组细胞凋亡率为(39.30±3.34)%(P<0.05),30、60、90mg·L^(-1)pPeOp组细胞凋亡率分别为(10.90±1.25)%、(28.80±2.70)%、(32.00±3.50)%,差异存在显著性(P<0.05)。TNF-R1、Fas/Fas L、caspase-3及caspase-8的m RNA和蛋白表达水平均上调,差异存在显著性(P<0.05);Bcl-2的m RNA和蛋白表达水平下调,差异存在显著性(P<0.05)。结论 pPeOp能明显抑制人胃癌细胞SGC-7901的增殖并诱导其凋亡,死亡受体通路和线粒体通路与pPeOp诱导人胃癌细胞SGC-7901凋亡有关。Aim To investigate the apoptosis mechanism of human gastric cancer cell SGC-7901 induced by Omphalia lapidescens protein pPeOp. Methods CCK-8 and flow cytometry were used to detect the inhibitory effect of different concentrations of pPeOp（ 30,60,90 mg·L^-1） on SGC-7901. The m RNA and protein expression of TNF-R1,Fas/Fas L,Bcl-2,caspase-3 and caspase-8 were detected by qRT-PCR and Western blot. Results SGC-7901 cells were treated with different concentrations of pPeOp（ 30,60,90 mg ·L^-1） for 24 h. CCK-8 test showed that there was no significant difference between PVP group and the control group. The survival rate of the 5-Fu group was（ 53. 71 ± 7. 34） %（ P〈0. 05）. The survival rates of pPeOp group（ 30,60,90 mg · L^-1） were（ 80. 95 ±6. 25） %,（ 53. 48 ± 5. 70） % and（ 44. 61 ± 6. 50） %（ r = 0. 984,P = 0. 016）,respectively. Flow cytometry showed that the apoptosis rate of PVP group had no significant difference with control group,and the apoptosis rate of 5-Fu group was about（ 39. 30 ± 3. 34） %（ P〈0. 05）. The apoptotic rates of pPeOp group（ 30,60,90 mg·L^-1） were（ 10. 90 ± 1. 25） %,（ 28. 80 ±2. 70） % and（ 32. 00 ± 3. 50） %,respectively（ P〈0. 05）. The m RNA and protein expression levels of Bcl-2 were down-regulated,whereas the expression of TNF-R1, Fas/Fas L, caspase-3 and caspase-8 were significantly up-regulated（ P〈0. 05）. Conclusions pPeOp can significantly inhibit the proliferation of gastric cancer cell line SGC-7901 and induce apoptosis in a dose-dependent manner. Death receptor pathway and mitochondrial pathway may be related to pPeOp-induced apoptosis of gastric cancer SGC-7901.

关 键 词：凋亡 CASPASES 胃癌 SGC-7901 线粒体通路 死亡受体通路 雷丸 

分 类 号：R282.71[医药卫生—中药学] R329.25[医药卫生—中医学]