重组p53腺病毒对人肺腺癌H1299细胞体内外的抑制作用  

Inhibitory effects of recombinant adenovirus-p53 on human lung adenocarcinoma H1299 cells in vitro and in vivo

作　　者：周小娟[1] 司小敏[1] 王云梅 吕建建[1] 

机构地区：[1]西安交通大学医学院第一附属医院肿瘤内科,陕西西安710061 [2]陕西省肿瘤医院内科,陕西西安710061

出　　处：《中国肿瘤生物治疗杂志》2013年第4期409-413,共5页Chinese Journal of Cancer Biotherapy

基　　金：陕西省科技计划资助项目(No.2010k14-02-01)~~

摘　　要：目的:研究重组p53腺病毒(recombinant adenovirus-p53,rAd-p53)在体内、外对肺腺癌H1299细胞(野生型p53基因缺失)生长的抑制作用,观察rAd-p53尾静脉注射治疗肺腺癌的可行性。方法:MTT法检测rAd-p53对H1299细胞增殖的抑制作用。rAd-p53以感染复数(multiplicity of infection,MOI)=500感染H1299细胞,24 h后RT-PCR检测H1299细胞中p53mRNA表达,72 h后Western blotting检测P53蛋白的表达、流式细胞术检测H1299细胞凋亡。H1299细胞皮下接种BALB/c裸鼠,建立裸鼠肺腺癌模型,尾静脉注射rAd-p53,观察肿瘤生长情况,绘制肿瘤生长曲线。结果:rAd-p53以MOI=500感染H1299细胞,24 h后有野生型p53 mRNA转录,72 h后有P53蛋白表达;且rAd-p53感染可明显抑制H1299细胞增殖,72 h时,rAd-p53组细胞增殖比显著低于对照组(2.8±0.4 vs 6.1±0.5,P<0.05)。感染rAd-p53后,随时间增加H1299细胞凋亡率呈上升趋势,48 h时rAd-p53组细胞凋亡率显著高于对照组,[(27.6±0.05)%vs(4.9±0.09)%,P<0.01]。成功建立H1299细胞荷瘤裸鼠模型,尾静脉注射rAd-p53 2周后移植瘤体积显著小于对照组[(0.875±0.253)vs(0.479±0.215)cm3,P<0.05]。结论:rAd-p53感染可上调H1299细胞P53蛋白的表达,抑制H1299细胞增殖、促进其凋亡,并且尾静脉注射rAd-p53可明显抑制H1299细胞裸鼠移植瘤的生长。Objective：To investigate the inhibitoty effects of recombinant adenovirus-p53 （rAd-p53） on the growth of lung adenocareinoma H1299 cells （wtP53^-/-） in vitro and in vivo, and observe the treatment feasibility of lung adenocarci- noma with tail intravenous injection of rAd-p53. Methods： MTT assay was performed to detect the inhibitory effect of rAd- p53 on the proliferation of H1299 cells. After transfected by rAd-p53 with multiplicity of infection （MOI） = 500, the ex- pression of p53 mRNA in H1299 cells was detected by RT-PCR at 24 h; the expression of P53 protein in H1299 cells and the apoptosis of H1299 cells were detected at 72 h by Western blotting and flow cytometry, respectively. BALB/c nude mice were injected subcutaneously with H1299 cells to establish a lung adenocarcinoma nude mice model and then the mice were intravenously administrated by rAd-p53; the tumor growth was observed and tumor growth curve was drawn. Results： H1299 cells were infected by rAd-p53 with MOI = 500; after infection for 24 h, wild-type p53 mRNA was ex- pressed in rAd-p53 group, and at 72 h, wt P53 protein was detected in rAd-p53 group, tAd-p53 infection could signifi- cantly inhibit the proliferation of H1299 cells, the cell proliferation ratio of rAd-p53 group was significant lower than that of the control group （2.8 ±0.4 vs 6.1 ±0.5, P 〈0. 05）. The apoptotic rates of H1299 cells in rAd-p53 group were increased with time, which were significantly higher than those in the control group （ [ 27.6 ±0.05 ] % vs [ 4.9 ± 0.09 ] %, P 〈 0.01 ） after infection for 48 h. H1299 tumor-bearing nude mice were successfully established, and the tumor volume of rAd-p53 group was significantly smaller than that of the control group even two weeks after tail intravenous injection （ [ 0. 875 ± 0. 253 ] cm3 vs [ 0. 479 ± 0. 215 ] cm3, P 〈 0.05 ）. Conclusion ： Tail intravenous infection of rAd-p53 could up-regulate the protein expression of P53 in H1299 and significantly inhibit the growth of H1299 cell xe ce

关 键 词：重组人P53腺病毒 肺腺癌 静脉注射 增殖 移植瘤 

分 类 号：R730.54[医药卫生—肿瘤] R734.2[医药卫生—临床医学]