胡桃醌诱导人宫颈鳞癌SiHa细胞凋亡机制  

Mechanism of apoptosis induced by juglone in human cervical cancer SiHa cells

作　　者：张巍[1] 朱文赫[1] 李妍[1] 罗军[1] 赵行宇[1] 徐俊杰[1] 姜艳霞[1] 吕士杰 

机构地区：[1]吉林医药学院,吉林吉林132013

出　　处：《中国药理学与毒理学杂志》2015年第5期831-835,共5页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金项目(21102055);吉林省科技厅科技发展项目(20130101157JC);吉林省教育厅十二五教育技术研究项目(2012-314);吉林省教育厅十二五教育技术研究项目(2014-371)~~

摘　　要：目的探讨胡桃醌对宫颈鳞癌Si Ha细胞促凋亡作用的机制,并研究其相关的信号转导通路。方法胡桃醌20μmol·L-1处理Si Ha细胞24,48和72 h后,倒置显微镜观察细胞形态学变化,MTT法检测细胞存活。胡桃醌20μmol·L-1处理Si Ha细胞24 h,流式细胞仪测定Si Ha细胞凋亡,Western蛋白印迹法检测凋亡相关蛋白BCL-2,BAX和激活型胱天蛋白酶3的表达,抑癌基因蛋白PTEN以及PI3K/AKT通路中AKT和p AKT的表达。结果与正常细胞对照组相比,胡桃醌组在24,48和72 h Si Ha细胞稀疏,脱壁,变圆;细胞存活抑制率分别为43.3%,63.0%和73.1%(P<0.05,P<0.01);胡桃醌20μmol·L-1处理细胞24 h后,细胞早期凋亡率增高至(6.47±1.79)%(P<0.05);BCL-2蛋白表达降低53.0%,BAX和激活型胱天蛋白酶3蛋白表达分别上升85.5%及183.3%,抑癌基因蛋白PTEN表达增加75.0%,p AKT表达降低45.8%(P<0.05)。结论胡桃醌可能通过上调PTEN的表达抑制PI3K/ATK通路,进而促进细胞凋亡。OBJECTIVE To explore the pro-apoptotic mechanism of juglone in Si Ha cells and to investigate its associated signal transduction pathways. METHODS Si Ha cells were treated with juglone 20 μmol·L-1for 24,48 and 72 h. Cellular morphology was detected by inverted microscopy.The cell viability was detected by methyl thiazolyl tetrazolium（MTT） assay. After 24 h treatment with juglone 20 μmol·L-1,the cell apoptosis was detected by flow cytometry while the expressions of apoptosis-related protein BCL-2,BAX and cleave-caspase-3,PI3K/AKt pathway-related protein PTEN,AKT and p AKT were detected by Western blotting. RESULTS After treatment with juglone for 24, 48 and72 h,the growth of Si Ha cells was significantly inhibited. Compared with cell control group,cells in juglone treated gruop were sparse,slipped off the wall,became round and the cell proliferation inhibitory rate was 43.3%,63.0% and 73.1%（P〈0.05,P〈0.01）,respectively. Twenty-four hours post treatment,the early apoptosis rate of juglone treated gruop cells was increased by（6.47±1.79）%（P〈0.01）compared with cell control group. Western blotting results showed that the expression of BCL- 2 decreased by53.0% while the expression of BAX and caspase-3 increased by 85.5% and 183.3%,respectively. The expression of PTEN was increased by 75.0% but the p AKt was decreased by 45.8%（P〈0.01）.CONCLUSION Juglone can upregulate the expression of PTEN, thus inhibiting PI3K/AKt pathway and promoting apoptosis of Si Ha cells.

关 键 词：胡桃醌 磷脂酰肌醇-3-激酶/丝氨酸/苏氨酶蛋白激酶 细胞凋亡 SIHA细胞 

分 类 号：R285[医药卫生—中药学]