重组survivin腺病毒对喉癌细胞抗凋亡作用的体外实验  

Influence of Anti-apoptosis Induced by Recombinant survivin Adenovirus in Laryngeal Cacinoma Cells

作　　者：吕元景[1] 苗素生[1] 贾深汕[1] 项丞[1] 何洪江[1] 刘伟松[2] 何国庆[1] 

机构地区：[1]哈尔滨医科大学附属第三医院头颈外科,哈尔滨150081 [2]海南省农垦总局医院口腔颌面外科

出　　处：《肿瘤防治研究》2011年第6期615-619,共5页Cancer Research on Prevention and Treatment

基　　金：黑龙江省青年科技基金资助项目(QC06C066);哈尔滨医科大学附属肿瘤医院科研启动基金资助项目(JJ2006-06)

摘　　要：目的观察survinvin基因对人喉癌细胞系HEP2的凋亡抑制作用及相关凋亡蛋白的表达变化。方法重组survivin腺病毒感染喉癌细胞系后分别用MTT法、FACS、Western法检测转染细胞生长情况,并应用表面增强激光解析离子化-飞行时间质谱仪(surface enhanced laser desorption/ionizationtime-of-flight mass spectrometry,SELDI-TOF-MS)分析蛋白质谱的变化。结果重组survivin腺病毒感染人喉癌Hep2细胞48h后,显微镜下可见明显的细胞生长旺盛;FACS检测见G1/S和G2/M期的细胞明显增多;Western法见凋亡相关蛋白CASPASE-3、CASPASE-10和CASPASE-11在病毒感染后表达减少;SELDI检测可见质荷比(m/z)分别为M4924_02、M8518_09和M2454_31的3个蛋白质峰在腺病毒感染组的峰度明显低于对照组。结论重组survivin腺病毒在体外能够有效地抑制人喉癌Hep2细胞凋亡,并且其作用与Caspase-3、Caspase-10和Caspase-11有相关性。Objective To observe apoptosis inhibition of survivin gene on laryngeal carcinoma cells Hep2 as well as to study protein profile changes by SELDI-TOF-MS（surface enhanced laser desorption/ionization time-of-flight mass spectrometry）in vitro.Methods Recombinant adenovirus vector of survivin was collected to infect laryngeal carcinoma cells Hep2.Its inhibiting effect on apoptosis was detected by MTT,FACS and Western.Infected cells were collected and cellular protein profile was analyzed by SELDI-TOF-MS.Results Two days after transfection,survivin adenovirus infected cells grew faster than those infected with blank adenovirus.FACS analysis showed that G1/S and G2/M phase cells remarkably increased with percentage of 26% of G2/M in survivin adenovirus infection.The expressions of apoptosis related proteins CASPASE-3,CASPASE-10 and CASPASE-11 decreased in the infected tumor cells.SELDI technique was used to detect cellular protein profile changes after infection.The peaks of M4924_02,M8518_09 and M2454_31 protein in SELDI decreased remarkably in the infection group.Conclusion survivin adenovirus could inhibit apoptosis in the human laryngeal carcinoma cell line Hep2,and there was a correlation with proteins Caspase-3,Caspase-10 and Caspase-11.

关 键 词：生存素 细胞凋亡 喉癌Hep2细胞 Caspase蛋白 SELDI-TOF-MS 

分 类 号：R739.65[医药卫生—肿瘤]