【摘要】 目的:研究miR-548a-3p靶向基质金属蛋白酶-2(MMP-2)抑制胃癌侵袭转移的分子机制。方法:通过生物信息学软件寻找可与MMP-2结合的miRNAs。qRT-PCR检测miR-548a-3p在不同分化和转移的胃癌细胞株中的表达,Western blot检测MMP-2表达;在三个重组细胞株中,qRT-PCR检测miR-548a-3p表达,Western blot检测MMP-2表达;双荧光素酶报告基因系统检测miR-548a-3p是否靶向结合MMP-2的3′-UTR端;Transwell实验检测转染前后胃癌细胞侵袭能力改变。结果:人低分化胃癌细胞株BGC-823中MMP-2高表达,而miR-548a-3p低表达;中、高分化人胃癌细胞株中MMP-2低表达,而miR-548a-3p高表达;双荧光素酶实验结果表明miR-548a-3p能靶向结合MMP-2的3′-UTR端;BGC-823-miR-548a-3p mimics细胞株中miR-548a-3p表达增高,而MMP-2表达下调,其他3个重组细胞株中miR-548a-3p表达水平较低,而MMP-2表达水平较高。BGC-823-miR-217 mimics侵袭细胞计数明显低于其他3组(P<0.05)。结论:miR-548a-3p能靶向结合MMP-2,下调其表达并抑制胃癌细胞的侵袭能力。更多还原

【Abstract】 Objective To study the mechanism of miR-548a-3p targeting MMP-2 inhibits the metastasisand invasion of gastric cancer cells. Methods Bioinformatics were used to search those miRNAs targeting MMP-2.The level of miR-548a-3p in gastric cancer tissues were detected by q RT-PCR and the expression of MMP-2 weredetected by Western blot. The level of miR-548a-3p in three reconstructed gastric cancer cell lines were detectedby qRT-PCR and the expression of MMP-2 were detected by Western blot. miR-548a-3p targeting the 3′-UTR ofMMP-2 were detected by Luciferase reporter assay detected. And the changes of ability of metastasis and invasionwere examined by Transwell experiment before and after transfection. Results There were high expression of miR-217 and low expression of MMP-2 in human poorly differentiated gastric cancer cell line BGC-823. There were lowexpression of miR-548a-3p and high expression of MMP-2 in human middle and high differentiated gastric cancercell line MKN-28 and SGC-7901(P < 0.05). The result of Luciferase reporter assay showed that miR-548a-3p cantargeting the 3`-UTR of MMP-2. There were high expression of miR-548a-3p and low expression of MMP-2 in MKN-28-miR-548a-3p mimics. There were low expression of miR-548a-3p and high expression of MMP-2 in othersreconstructed three cell lines. There were smaller invaded cells in SGC-7901-miR-217 mimics than in others threecell lines(P < 0.05). Conclusions miR-548a-3p can target the 3′-UTR of MMP-2 and down regulate itsexpression and inhibit the ability of invasion of gastric cancer cells.更多还原