RNA干扰PTTG基因对泌乳素型垂体瘤细胞凋亡的机制研究  

Mechanism study of PTTG on the apoptosis of human prolactin pituitary adenoma

作　　者：张静[1] 王秋月[1] 刘峥嵘[2] 姜雅秋[1] 刘洁[3] 崔泽实[3] 

机构地区：[1]中国医科大学附属第一医院内分泌科,辽宁沈阳110001 [2]辽宁省人民医院普外科,辽宁沈阳110016 [3]中国医科大学实验技术中心,辽宁沈阳110001

出　　处：《中国现代医学杂志》2010年第14期2104-2108,2114,共6页China Journal of Modern Medicine

基　　金：辽宁省博士启动基金(No:20051039)

摘　　要：目的研究RNA干扰垂体瘤转化基因(PTTG)对泌乳素型垂体瘤细胞(HPA)凋亡的影响并探讨其作用机制。方法设计并合成靶定PTTG的特异性siRNA片段,转入体外培养HPA细胞,48h后验证siRNA的干扰效应;Annexin V/PI染色法观察PTTG基因沉默对细胞凋亡的影响;RT-PCR和Western blot检测caspase-3在mRNA和蛋白水平表达;caspase活性检测试剂盒检测caspase活性。结果设计合成的PTTGsiRNA转染后,能够有效抑制体外培养HPA细胞内PTTG表达实现其基因沉默效应;经浓度梯度CDDP诱导24h,转染和未转染细胞均产生细胞凋亡,caspase-3表达增强,并呈现出浓度依赖性;RNA干扰组细胞凋亡率低于未转染组,转染组caspase-3活化受抑,其caspase-3基因表达、蛋白活性均低于未转染组(P<0.05)。结论 PTTG在人泌乳素型垂体瘤中过表达,能通过激活caspase-3促进HPA细胞凋亡,可作为该肿瘤临床诊断和基因治疗的新靶点。【Objective】To investigate the effect of siRNA targeting pituitary tumor transforming gene（PTTG） on cell apoptosis of cultured human prolactin pituitary adenoma in vitro and it＇s mechanism.【Methods】 siRNA targeting PTTG was designed and composed,transfected into the cultured HPA cells,cell apoptosis was detected by Annexin V/PI staining kit.RT-PCR and western blot technique were used to test caspase-3 expression.The activity of caspase-3 was determined by a caspases colorimetric assay kit.【Results】The PTTG siRNA designed could interfere the PTTG expression of HPA.After CDDP induction for 24hours,HPA appeared cell apoptosis and caspase-3 increased.The apoptotic rate and caspase-3 activation level decreased in siRNA transfection group.【Conclusions】PTTG gene may induce cell apoptosis in human prolactin pituitary adenoma by up-regulating the expression of caspase-3.It could be regarded as a novel target for clinical diagnosis and gene therapy.

关 键 词：垂体瘤转化基因 siRNA 人泌乳素型垂体瘤 细胞凋亡 CASPASE-3 

分 类 号：R739.41[医药卫生—肿瘤]