携带凋亡素基因的重组腺病毒对人肺鳞癌细胞SK-MES-1、人肺腺癌细胞NCI-H1299凋亡的影响  

Effects of Recombinant Adenovirus Carrying Apoptin Gene on the Apoptosis of Human Lung Squamous Carcinoma SK-MES-1 Cell Lines and Human Lung Adenocarcinoma NCI-H1299 Cell Lines

作　　者：阮军[1] 吴友茹[1] 缪李丽[2] 

机构地区：[1]绵阳市中心医院呼吸内科,四川绵阳621000 [2]重庆医科大学附属永川医院呼吸内科,重庆402160

出　　处：《中国药房》2015年第22期3066-3069,共4页China Pharmacy

基　　金：重庆市医学科研计划项目(No.2011-1-084)

摘　　要：目的:研究携带凋亡素基因VP3的重组腺病毒载体Adxsi-GFP-VP3对人肺鳞癌细胞SK-MES-1、人肺腺癌细胞NCI-H1299的凋亡的影响。方法:取对数生长期SK-MES-1、NCI-H1299细胞,各分为重组腺病毒(Adxsi-GFP-VP3)组、空病毒(Adxsi-GFP)组和细胞对照(磷酸盐缓冲液)组,依次记为A、B、C组。采用逆转录-聚合酶链反应和Western blot法检测A、B组细胞转染48、72 h的VP3 mRNA和Apoptin表达情况,透射电镜观察A组细胞转染72 h的超微结构变化。采用MTT法检测3组细胞转染24、48、72、96 h的细胞增殖活性;流式细胞术检测细胞转染后24、48、72 h的凋亡率及细胞周期变化。结果:与B组比较,A组SK-MES-1、NCI-H1299细胞转染48 h后有VP3 mRNA表达,转染72 h后有Apoptin表达。A组转染72 h后SK-MES-1、NCI-H1299细胞均出现细胞凋亡的超微结构改变。与B、C组比较,A组SK-MES-1、NCI-H1299细胞增殖活性降低、细胞凋亡率增加,且与转染时间呈正相关;转染72 h S期的SK-MES-1、NCI-H1299细胞比例降低、G2/M期细胞比例增加。以上差异均具有统计学意义(P<0.05)。结论:Adxsi-GFP-VP3能有效诱导SK-MES-1、NCI-H1299细胞凋亡。OBJECTIVE：To explore the effects of recombinant adenovirus vector Adxsi-GFP-VP3 carrying apoptin gene VP3 on the apoptosis of human lung squamous carcinoma SK-MES-1 cell lines and human lung adenocarcinoma NCI-H1299 cell lines.METHODS：The exponential phase SK-MES-1 and NCI-H1299 cell lines were respectively divided into a recombinant adenovirus（Adxsi-GFP-VP3）group,a empty virus（Adxsi-GFP）group and a cell control（phosphate buffer）group,which were marked as group A,B and C respectively.Reverse transcription polymerase chain reaction and Western blot method were used to detect the expressions of VP3 mRNA and Apoptin in the cells of groups A and B 48 and 72 h after transfection.The change in the ultrastructure of the cells in group A was observed under transmission electron microscope 72 h thereafter.MTT method was adopted to detect the cell proliferation activities of three groups 24,48,72 and 96 h thereafter and flow cytometry to determine the apoptosis rates and cell cycle changes 24,48 and 72 h thereafter.RESULTS：Compared to group B,group A demonstrated the expression of VP3 mRNA in SK-MES-1 and NCI-H1299 cell lines 48 h after transfection,and Apoptin expression and ultrastructure change for apoptosis of SK-MES-1 and NCI-H1299 cell lines 72 h thereafter.Compared to groups B and C,group A showed lower proliferation activities and higher apoptosis rates of SK-MES-1 and NCI-H1299 cell lines,which had a positive correlation with transfection time;and in the group A,there was a decrease in the proportion of the SK-MES-1 and NCI-H1299 cell lines in S phase and an increase in the proportion of those in G2/M phase,72 h after transfection.There was statistically difference（P〈0.05）.CONCLUSIONS：Adxsi-GFP-VP3 can effectively induce the apoptosis of SK-MES-1 and NCI-H1299 cell lines.

关 键 词：凋亡素基因VP3 重组腺病毒载体Adxsi-GFP-VP3 人肺鳞癌细胞SK-MES-1 人肺腺癌细胞NCI-H1299 蛋白表达 凋亡 细胞周期 

分 类 号：R965[医药卫生—药理学]