腺病毒介导的ING4基因对SMMC-7721人肝癌细胞生长的抑制效应  

The Growth Inhibitory Effect of Ad-ING4 on Human Hepatocarcinoma Cell Line SMMC-7721

作　　者：杨苾雯[1] 曹婧媛[1] 徐俊[1] 方悦之[1] 王玲慧[1] 姚雷[1] 杨吉成[1] 

机构地区：[1]苏州大学医学部细胞与分子生物学系,江苏苏州215123

出　　处：《苏州大学学报（医学版）》2010年第5期926-930,F0003,共6页Suzhou University Journal of Medical Science

基　　金：2009年苏州大学国家大学生创新性实验计划项目(57315733)

摘　　要：目的研究腺病毒介导的ING4(Ad-ING4)基因对体外SMMC-7721人肝癌细胞的生长抑制作用。方法将腺病毒空载体Ad-GFP及重组腺病毒Ad-ING4分别感染SMMC-7721细胞,采用RT-PCR法及间接免疫荧光检测ING4基因的转录和表达;荧光显微镜观察Ad-ING4对SMMC-7721细胞的细胞毒作用;MTT比色法绘制细胞生长曲线,计算生长抑制率;流式细胞术(FCM)检测Ad-ING4对细胞凋亡的影响;DAPI核荧光染色检测细胞凋亡的核形态学变化。结果 RT-PCR及间接免疫荧光显示Ad-ING4能在SMMC-7721细胞内稳定表达;Ad-ING4感染72h和96h后对细胞的生长抑制率达33.82%和50.70%;FCM检测有明显的凋亡峰,凋亡率可达46.7%;DAPI染色结果显示SMMC-7721细胞胞核呈现核深染、固缩、断裂等细胞核凋亡的形态改变。结论 Ad-ING4能够明显抑制SMMC-7721细胞的增殖,并诱导其凋亡。Objective To investigate the effect of adenovirus-mediated ING4 gene （ Ad-ING4） on human hepatocarcinoma cell line SMMC-7721 in vitro. Methods After the Ad-ING4 was transfected into SMMC-7721 cells,we used RT-PCR and indirect immunofluorescence to confirm that ING4 gene transcripted and expressed in SMMC-7721 cells. Fluorescent microscopy was used to observe the cytotoxicity of Ad-ING4 to SMMC-7721 cells. The proliferation curve of SMMC-7721 cells was drawn based on MTT assay,and the inhibitory rate was calculated. Cellular Apoptosis of SMMC-7721 cells induced by Ad-ING4 was detected by FCM. DAPI staining was used to observe the morphological changes of the nuclei. Results The transcription and expression of ING4 gene in SMMC-7721 cells was detected by RT-PCR and indirect immunofluorescence. Seventy-two hours and ninety-six hours after infecting with Ad-ING4,the growth inhibitory rate was 33. 82% and 50. 70% respectively. The apoptosis peak was found by FCM and the rate could reach 46. 7% . The classical morphological changes of the nuclei were shown by DAPI staining such as anachromasis,karyopycnosis,breakage and so on. Conclusion Ad-ING4 could significantly inhibit the proliferation of SMMC-7721 cells,and induce the cells to apoptosis.

关 键 词：AD-ING4 SMMC-7721细胞 生长抑制 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤] R730.59[医药卫生—临床医学]