HP联合MNNG对人食管上皮细胞周期及增殖影响  

Effects of HP combined with MNNG on cycle and proliferation of human esophageal epithelial cells

作　　者：雷毅 张晨 李思瑶 陈艳[2] LEI Yi;ZHANG Chen;LI Siyao;CHEN Yan(Department of Health Toxicology,College of Public Health,Xinjiang Medical University,Urumqi 830011,China;The Medical School of Jiaxing University,Jiaxing Zhejiang 314000,China)

机构地区：[1]新疆医科大学公共卫生学院卫生毒理学教研室,乌鲁木齐830011 [2]嘉兴学院医学院,浙江嘉兴314000

出　　处：《新疆医科大学学报》2022年第5期492-498,共7页Journal of Xinjiang Medical University

基　　金：浙江省自然科学基金(LY20H260006)。

摘　　要：目的HP联合MNNG处理人食管上皮(HEEC)细胞,观察其对细胞增殖及周期的影响。方法CCK8法测定MNNG的IC_(50)值,确定染毒浓度;HP以感染复数(MOI)=100∶1(细菌∶细胞=100∶1)为实验浓度;分为对照组、HP处理组、MNNG处理组、HP+MNNG联合处理组(联合组),并分别在处理后1、6、12 h测定细胞增殖能力、细胞周期及相关周期蛋白表达的变化。结果MNNG IC_(50)值为40.90μmoL/L,本研究以1/2 IC_(50)值为实验浓度。对照组和MNNG组细胞形态无明显变化,在6 h和12 h后HP处理组和联合组细胞形态有较大改变,主要表现为细胞质紊乱不均匀,出现空泡样变化等。在经过1、6、12 h处理后,各组增殖速度为联合组<HP处理组<MNNG处理组<对照组(F_(1 h趋势)=1089.3,P<0.001;F_(6 h趋势)=322.78,P<0.001;F_(12 h趋势)=50.01,P<0.001)。各处理组随着时间的延长,细胞增殖速度变缓(F_(HP)=370.44,P<0.001;F_(MNNG)=65.01,P<0.001;F_(联合)=98.46,P<0.001)。各组细胞在不同时间处理后G0/G1、S、G2/M细胞周期差异均有统计学意义(P<0.05)。随着染毒时间的延长,HP组、MNNG组和联合组均呈现G2/M期阻滞逐渐增大(F_(HP)=238.76,P<0.001;F_(MNNG)=182.47,P<0.001;F_(联合)=58.43,P<0.001),且同一时间为联合组大于HP和MNNG单独处理组(F_(1 h)=32.73,P<0.001;F_(6 h)=70.37,P<0.001;F_(12 h)=125.41,P<0.001)。染毒12 h后与对照组相比,其余3组细胞Cyclin B1蛋白表达量均显著升高,差异有统计学意义(P均<0.001),以联合组Cyclin B1表达最高,差异有统计学意义(P均<0.001)。结论HP及MNNG均能抑制HEEC细胞的增殖并改变其固有的细胞周期,联合作用表现更为明显,周期蛋白Cyclin B1表达升高,细胞周期G2/M期阻滞增大。Objective To observe the effect of cell proliferation and cell cycle with HP combined with MNNG on human esophageal epithelial(HEEC)cells on cell proliferation and cycle.Methods IC_(50) value of MNNG was determined by CCK8 method,and exposure concentration was determined;HP took multiplicity of infection(MOI)=100∶1(bacteria∶cells=100∶1)as the experimental concentration;divided into normal control group,HP treatment group,MNNG treatment group group,HP+MNNG combined treatment group,and the changes of cell proliferation,cycle and related cyclin expression were measured at 1,6,and 12 hours after the treatment,respectively.Results IC_(50) value of MNNG was 40.90μmoL/L,and 1/2 of IC_(50) value was used as the experimental concentration in the study.The morphology of cells in the control group and the MNNG group did not change significantly.After 6 hours and 12 hours,the morphology of cells in the HP treatment group and the combined treatment group changed greatly,it mainly manifested as cytoplasmic disorder and vacuolar-like changes.After 1,6,and 12 hours of the treatment,the proliferation rate of each group was combination group<HP<MNNG<control group(F_(1 h trend)=1089.3,P<0.001;F_(6 h trend)=322.78,P<0.001;F_(12 h trend)=50.01,P<0.001).With the prolongation of time,cell proliferation rate of the each treatment group was decreased(F_(HP)=370.44,P<0.001;F_(MNNG)=65.01,P<0.001;F_(combined group)=98.46,P<0.001).The G0/G1,S,G2/M phases of cell cycle of the each group of cells after the treatment at different times were statistically significant(P<0.05).With the prolongation of exposure time,G2/M phase arrest of cell cycle was gradually increased in HP group,MNNG group and combined treatment group(F_(HP)=238.76,P<0.001;F_(MNNG)=182.47,P<0.001;F_(combined group)=58.43,P<0.001).The combined group was greater than the HP and the MNNG groups alone(F_(1 h)=32.73,P<0.001;F_(6 h)=70.37,P<0.001;F_(12 h)=125.41,P<0.001).12 hours after exposure,compared with the control group,the expression of Cyclin B1 protein in the treatme

关 键 词：HP MNNG 联合作用 HEEC细胞 细胞周期 

分 类 号：R735.1[医药卫生—肿瘤]