不同孕期小鼠羊水对小鼠肝癌细胞H22增殖和凋亡的影响  

Effects of mouse amniotic fluid collected on different gestational days on proliferation and apoptosis of H_(22) cells in vitro

作　　者：丁雅明[1] 方廖琼[1] 周兰[1] 张弘[1] 白晋[1] 王智彪[1] 

机构地区：[1]重庆医科大学生物医学工程系,重庆医科大学医学超声工程研究所,重庆400016

出　　处：《第三军医大学学报》2008年第12期1136-1139,共4页Journal of Third Military Medical University

基　　金：国家教育部创新团队发展计划(2005-33);国家人事部留学人员科技活动项目择优资助(2005-85)~~

摘　　要：目的观察不同孕期小鼠羊水对小鼠肝癌细胞H22增殖和凋亡的影响。方法用孕11、13、15 d和17 d的KM小鼠羊水-RPMI1640培养基培养H22细胞(培养液中羊水终浓度为10%),RPMI1640培养基培养的H22细胞作为对照组,采用MTT法检测细胞的增殖能力;Annexin V-FITC法检测细胞凋亡率,透射电镜观察细胞形态;S-P免疫荧光染色法经激光共聚焦显微镜分析H22细胞中增殖细胞核抗原(PCNA)和p53的表达。结果羊水能明显抑制H22细胞的体外增殖和PCNA的表达,随孕期的延长而增高(P<0.05);并能促进p53的表达以及诱导H22细胞凋亡,随孕期的延长而降低(P<0.05)。同时,PCNA和p53的表达强度具有明显的负相关性(r=-0.808,P<0.01)。结论孕11、13、15、17 d的小鼠羊水能抑制H22细胞增殖,诱导其凋亡,可能与PCNA和p53的表达有关,并且孕期越早、干预时间越长,效果越显著。Objective To investigate the effects of mouse amniotic fluid on the proliferation and apoptosis of mouse hepatoma cell line H22. Methods Treated with the mixture of RPMI 1640 medium and 10% mouse amniotic fluid collected respectively on gestational day 11, 13, 15 and 17 or RPMI 1640 medium as control, the proliferation of H22 ceils was detected with MTT and the apoptosis of H22 ceils with Annexin V-FITC; H22 cells were observed morphologically under electron microscope; The expressions of proliferating cell nuclear antigen （PCNA） and p53 in H22 cells were investigated with immunofluorescence staining （ S-P method） and laser confocal microscopy. Results Amniotic fluid significantly inhibited the cell proliferation as well as the PCNA expressions, increased the p53 expressions and induced cell apoptosis. The amniotic fluid collected on gestational day 11 was the most effective, while that of gestational day 17 was the least effective （ P 〈 0. 05 ）, and their effects were both in a time-dependent manner. There was a negative correlation between PCNA and p53 （ r = -0.808, P 〈0. 01 ）. Conclusion Amniotic fluid collected on gestational day 11, 13, 15 and 17 can inhibit the proliferation of H22 ceils and induce their apoptosis. The earlier pregnancy and the longer intervention is, more significant effects of amniotic fluid are.

关 键 词：H22细胞 羊水 凋亡 增殖细胞核抗原 P53 

分 类 号：R73-354[医药卫生—肿瘤] R73-36[医药卫生—临床医学]