微RNA-101调控人胃癌细胞系增殖、迁移、侵袭的体内外研究  

In vitro and in vivo study on the regulation of microRNA-101 in human gastric cancer proliferation, migration and invasion

作　　者：杜静[1] 黄伟[1] 陈有为[1] 阮洪军[1] 

机构地区：[1]浙江省人民医院消化内科,杭州310014

出　　处：《中华消化杂志》2015年第8期510-515,共6页Chinese Journal of Digestion

摘　　要：目的了解miRNA-i01在胃癌组织和细胞中的表达水平，以及对胃癌细胞增殖、迁移、侵袭力和皮下移植瘤的影响。方法采用实时定量PCR分析28份胃癌组织与人胃癌细胞系BGC-823、SGC-7901、MKN-45、ACTS，以及人胃黏膜上皮细胞系GES-1中miRNA-101的表达水平。构建miRNA-101重组腺病毒表达载体。应用MTT法检测miRNA—101对胃癌细胞增殖能力的影响。应用Millipore Transwell小室法检测胃癌细胞的迁移和侵袭能力。使用BALB／c裸鼠建立胃癌皮下移植瘤模型，并比较肿瘤体积。统计学分析采用t检验。结果胃癌组织中miRNA一101的表达量为0．661±0．396，低于所对应的癌旁组织中的1．128±0．697，差异有统计学意义（t=10．091，P〈0．01）。胃正常上皮细胞GES-1中的miRNA-101表达量高于胃癌细胞系BGC-823、SGC-7901、MKN-45和AGS。miRNA—101对MKN-45细胞的增殖能力呈现明显的抑制作用，对胃癌细胞BGC-823、SGC-7901、MKN-45、AGS的迁移和侵袭均有抑制作用。选取MKN-45细胞系建立裸鼠皮下移植瘤模型后5周，miRNA-101重组腺病毒（Ad-miRNA-101）组肿瘤体积为（333．56±46．71）mm^3，小于空腺病毒一增强绿色荧光蛋白（Ad—EGFP）组的（806．41±51．83）mm^3，差异有统计学意义（t=21．431，P〈0．01）。结论在胃组织和细胞中，miRNA-101是一个抑制性mIRNA，其表达水平下调参与了胃癌的发生和发展，有可能成为胃癌生物靶向治疗的新靶点。Objective To investigate the expression of microRNA-101 （miRNA-101） in human gastric cancer, and to explore its effects on proliferation, migration and invasion of gastric cancer cell. Methods The expression of miRNA-101 in 28 human gastric cancer tissues, human gastric cell lines BGC-823, SGC-7901, MKN-45, AGS and human normal gastric epithelial cell line GES-1 were determined by real time polymerase chain reaction （PCR）. Recombinant miRNA-101 adenovirus vector was constructed. The effects of miRNA-101 on gastric cancer proliferation was detected with cell proliferation assay. The ability of gastric cancer cell migration and invasion was assessed with Transwell assay. Gastric xenograft cancer model was established in BALB/c nude mice and the tumor size was compared. The t test was used for the statistical analysis. Results The expression of miRNA-101 in gastric cancer tissues was 0. 661 ±0. 396, which was lower than that of corresponding para carcinoma tissues （1. 128 ±0. 697）, and the difference was statistically significant （t = 10. 091, P〈0.01）. The expression of miRNA-101 in normal gastric epithelial cell line GES-1 was higher than those of gastric cancer cell lines BGC-823, SGC-7901, MKN-45 and AGS. There was significant suppression role of miRNA-101 on MKN-45 cells proliferation, and which also had inhibition role on cell migration and invasion of gastric cancer cell lines BGC-823, SGC-7901, MKN-45 and AGS. At five weeks after MKN-45 gastric xenograft cancer nude mice model established, the tumor size of Ad-miRNA-101 group （（333.56±46, 71） mm^3） was smaller than that of Ad-enhanced green fluorescent protein （EGFP） group （806.41± 51.83） mm^3, and the difference was statistically significant （t = 21. 431, P〈 0. 01）. Conclusion In gastric tissues and cells, miRNA-101 is a tumor suppressive miRNA and its downregulated expression involved in the genesis and development of gastric cancer, which may be a new target of biological target therapy in gastric cancer.

关 键 词：微RNAS 胃肿瘤 miRNA-101 

分 类 号：R735.2[医药卫生—肿瘤]