微小RNA-375对胃癌细胞的功能研究  

Study on the effects of microRNA-375 on gastric cancer cell function

作　　者：张莘悦[1] 张伟[1] 陈卫昌[1] 

机构地区：[1]苏州大学附属第一医院消化科,江苏省215006

出　　处：《中华消化杂志》2013年第8期518-522,共5页Chinese Journal of Digestion

摘　　要：目的探讨微小RNA（miRNA）375对胃癌细胞的功能研究，并讨论其可能的作用机制。方法将miRNA-375的真核表达载体稳定转染人人胃癌细胞株HGC-27，筛选并建立高表达miRNA-375的胃癌细胞（PC3．1-375）组。另设空白质粒转染组（PC3．1组）和空白对照组。通过细胞计数试剂盒8（cCK-8）法和流式细胞检测法检测3组细胞的增殖能力和凋亡率。Western印迹法检测3组细胞中星形胶质细胞上调基因-l（AEG-1）蛋白表达。建立胃癌裸鼠皮下种植瘤模型，种植30d后处死裸鼠取瘤测量。免疫组织化学法检测移植瘤组织内AEG-1的蛋白表达。统计学方法采用t检验和方差分析。结果PC3．1-375组细胞的miRNA-375的表达量较空白对照组和PC3．1组明显上升（￡-2．82、2．01，P均d0．05），同时增殖能力显著降低，实验第3天时差异最大（t。d=3．56、2．79，P均d0．05），凋亡率显著升高（t=2．05、2．96，P均d0．05）。Western印迹结果示，AEG-1蛋白在PC3 1-375组中的表达较空白对照组和PC3．1组下降（t=3．78、2．26，P均d0．05）。至实验终点时，空白对照组裸鼠皮下种植瘤的体积和质量分别为（3．12±0．77）cm。和（3．86±0．56）g，PC3．1组为（3．76±0．65）cm。和（4．01±0．52）g，均显著大于PC3．1-375组的（0．72±0．21）cm。和（0．85±0．15）g（体积：t=5．67、6．92，质量：t=4．56、5．01，P均〈0．05）。AEG-1蛋白在PC3．1-375组种植瘤中的表达较空白对照和PC3．1组明显下调（F=35．000，P〈O．05）。结论miRNA375可有效抑制胃癌细胞HGC-27的增殖，促进细胞凋亡，miRNA375在胃癌中是-种抑瘤因子。AEG-1可能是miRNA-375作用的靶基因。Objective To investigate the effects of microRNA（miRNA）-375 on gastric cancer cell function and the possible mechanism. Methods miRNA 375 eukaryotic expression vector was stably transfected into human gastric cancer cell line HGC-27. A miRNA-375 highly expressed gastric cancer cell line PC3. 1-375 was screened and established. Blank plasmid transfection group （PC3. 1group） and blank control group were also established. The cell proliferation ability and apoptosis rate in three groups were detected by cell counting kit-8 （CCK-8） method and flow cytometry. The protein expression of astrocyte elevated gene-1 （AEG-1） in three groups was determined by Western blotting. The nude mouse subcutaneous xenografts model was also established. After 30 days, the nude mice were executed and the xenografts were dissociated and measured. The expression of AEC-I in the tumors at protein level was tested by immunohistochemistry method. The statistical inference includes t-test and analysis of variance. Results After stable transfection and cell screening, compared with that of blank control group, the expression of miRNA-375 in PC3.1-375 group significantly increased （t= 2.82, 2.01, bothP〈0.05）. The cell proliferation of PC3. 1-375 group was significantly lowerthan that of PC3. 1 group and blank control group and the difference reached a maximum in day 3 （t=3.56, 2.79, both P〈0.05）. But with the apoptosis rate of PC3.1-375 group, the opposite was true （t=2.05, 2.96, both P%0.05）. The results of Western blot shows that the expression of AEG- 1 at protein level of PC3.1-375 group decreased compared with that of blank control group and PC3.1 group. The xenografts growth rate, average weight and tumor volume of PC3. 1 375 group were significantly lower than those of blank control group and PC3. 1 group （t=3. 78, 2. 26, both P〈0.05）. By the end of experiment, the volume of nude mouse subcutaneous xenografts in blank control group （（3.12± 0.77） cm3） and PC3.1 group （（3.76 ±0.65） cma�

关 键 词：胃肿瘤 微RNAS 转染 细胞增殖 细胞凋亡 疾病模型 动物 肿瘤种植 

分 类 号：R735.2[医药卫生—肿瘤]