人乳头状瘤病毒转染对甲基硝基亚硝基胍致人食管上皮Het-1A细胞增殖及凋亡的影响  

Effect of human papillomavirus transfection on MNNG induced cell proliferation and apoptosis of human esophageal epithelial cell line Het-1A

作　　者：李仪楠 马月[1] 郑雨虹 刘冉[1] 浦跃朴[1] 尹立红[1] LI H-nan MA Yue ZHENG Yu-hong LIU Ran PU Yue-pu YIN Li- hong(Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, Jiangsu 210009, China）.)

机构地区：[1]东南大学公共卫生学院环境医学工程教育部重点实验室,江苏南京210009

出　　处：《环境与职业医学》2017年第6期490-495,共6页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金项目(编号:81573108;81573191)

摘　　要：[目的]研究转染人乳头状瘤病毒18型(HPV18)后,甲基硝基亚硝基胍(MNNG)暴露对人食管上皮Het-1A细胞周期和增殖、凋亡功能的影响。[方法]通过慢病毒介导稳定转染HPV18 E6、E7基因到人食管上皮Het-1A细胞中,荧光定量PCR检测E6和E7相对表达量后得到稳定表达株。应用MTT法观察MNNG(0、1、5、10μmol/L)染毒处理24 h后对转染组和对照组细胞增殖功能的影响。应用流式细胞术检测转染组和对照组细胞染毒后细胞周期和凋亡的变化。[结果]HPV18转染组与对照组相比,细胞增殖能力增强,G1期延长,S期缩短(P<0.05),且凋亡率下降(P<0.05)。非转染细胞中MNNG染毒各组与未染毒组相比,细胞增殖活性受到抑制,G1期延长,细胞凋亡率增加(P<0.05)。HPV18转染后MNNG暴露,可导致G2期延长;MTT检测细胞增殖结果显示HPV18转染组细胞在MNNG作用下,细胞存活率均高于同浓度MNNG单独暴露的非转染组细胞;中、低浓度MNNG暴露后HPV18转染组较未转染组相比细胞凋亡率降低(P<0.05)。[结论]Het-1A细胞转染HPV18后暴露MNNG,细胞凋亡受到一定程度抑制,促进细胞增殖。由此可能导致细胞不断积累MNNG引起的基因突变和DNA损伤。[ Objective ] To study N-methyl-N＇-nitro-N-nitroso-guanidine （MNNG） induced cell proliferation and apoptosis of human esophageal epithelial cell line Het-1A with human papillomavirus （HPV） 18 transfection. [ Methods ] After transfecting lentiviral vectors of HPV18 E6 and E7 genes in Het-lA cells, the expressions of E6 and E7 were identified by fluorescence quantitative PCR. Both transfected group and control group were exposed to MNNG （0, 1, 5, and 10 lamol/L） for 24 h. MTT assay was applied to detect cell growth inhibition. Flow cytometry was processed to investigate the changes of cell cycle and apoptosis induced by MNNG （0, 1, 5, and 10 pmol/L） in HPV18 transfected group and control group. [ Results ] In the HPV18 transfected group, cell proliferation was promoted, G1 phase was extended, S phase was shortened （P〈 0.05）, and the apoptosis rate was reduced （P 〈 0.05） compared with the control group. In the control group, MNNG lengthened ceils in G1 phase, and cell apoptosis was increased （P〈0.05）. In the transfected group with MNNG exposure, G2 phase was elongated compared with exclusive exposure to MNNG. Combined effects of HPV18 and MNNG increased the cell viability rate compared withthe effect of exclusive exposure to MNNG at the same concentration. Combined effect of HPV18 and low or medium concentrations of MNNG reduced the apoptosis rate compared with exclusive exposure to MNNG（P 〈 0.05）. [ Conclusion ] Het-lA cells with HPV18 transfection and exposure to MNNG result in cell apoptosis inhibition, cell proliferation increase, gene mutation, and DNA damage.

关 键 词：人乳头状瘤病毒18 甲基硝基亚硝基胍 人食管上皮Het-1A细胞 

分 类 号：R114[医药卫生—卫生毒理学]