摘要: |
目的 通过实验研究miRNA-101在胃癌中的表达水平,以及对胃癌细胞增殖、迁移、侵袭力和皮下移植瘤的影响,以探讨将miRNA-101用于胃癌生物靶向治疗的可能性。方法 分子克隆技术构建miRNA-101重组腺病毒表达载体,MTT法检测细胞增殖能力,MilliporeTranswell小室法检测细胞迁移能力,Matirgel法检测细胞侵袭能力,q-PCR定量分析各基因表达水平,使用BALB/c免疫缺陷裸鼠建立胃癌皮下移植瘤模型。结果miRNA-101在胃癌组织中的表达量为(0.661±0.396),低于所对应的癌旁组织(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,Ad-miRNA-101组肿瘤体积为(333.56±46.71)mm3,小于Ad-EGFP组(806.41±51.83)mm3,差异有统计学意义(t=21.431,P<0.01)。结论在胃组织中,miRNA-101是一个抑制性miRNA,miRNA-101表达水平下调参与胃癌的发生和发展。miRNA-101将有可能成为胃癌生物靶向治疗的新靶点。 |
关键词: microRNA 胃癌 miRNA-101 |
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基金项目:MicroRNA Gastric cancer miRNA-101 |
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Effect of MicroRNA-101 on cell proliferation,migration and invasion of human gastric cancer cell lines |
WEI Furong,RUAN Hongjun,WANG Huiju,HE Xujun,DU Jing,MA Jie
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Zhejiang provincial People's Hospital
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Abstract: |
Objective To investigate the effect of miRNA-101 on cell proliferation, migration and invasion of human gastric cancer cells in vitro and in vivo. Methods Recombinant adenovirus encoding miRNA-101 was constructed, MTT assay was to detect vitality and proliferation ability of cells, transmembrane motility assay was for observing cell migration and invasion, The target gene expression was analyzed using real time PCR. BALA/c nude mice were injected to develop the gastric xenograft cancer model. Results miRNA-101 expression was significantly lower in gastric cancer tissue (0.661 ±0.396) than that in matched normal tissues (1.128±0.697, P<0.05); and lower in BGC-823, SGC-7901, MKN-45, AGS gastric cancer cells than that in normal gastric epithelial GES-1 cells. Reestablishment of miRNA-101 was able to significantly inhibit the proliferative abili- ty of MKN-45 cells, and induce the reduction of cellular proliferation, migration and invasion in BGC-823, SGC-7901, MKN-45 and AGS cellls in vitro. The xenograft model further confirmed that miRNA-101 massively suppresses the tumor growth in vivo (333.56±46.71 mm3 vs 806.41±51.83mm3). Conclusion Down-regulation of miRNA-101 may be involved in tumorigenesis of the stomach, and it might bea potential target for a novo therapeutic approach of gastric cancer. |
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