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作 者:杨屹立[1] 何学元[1] 潘新民[1] 马建勋[1] YANG Yi-li;HE Xue-yuan;PAN Xin-min;MA Jian-xun(Department of General Surgery, Gansu Provincial People's Hospital, Lanzhou 730000, China)
出 处:《基础医学与临床》2020年第8期1068-1075,共8页Basic and Clinical Medicine
基 金:甘肃省科技计划(18JR3RA330);甘肃省人民医院院内科研基金(18GSSY4-44)。
摘 要:目的探讨miR-205-5p靶向RAS致癌家族基因2B(RAP2B)对胃癌细胞增殖、迁移和侵袭的影响。方法RT-qPCR和Western blot检测胃癌细胞AGS、MGC803、MKN-28、SGC-7901和正常胃黏膜细胞GES-1中miR-205-5p和RAS致癌家族基因2B的表达。分别构建过表达miR-205-5p和抑制RAP2B表达的AGS细胞株,采用MTT法检测细胞活力;Transwell小室法检测细胞的迁移及侵袭能力;Western blot检测RAP2B、cyclin D1、MMP-2、MMP-9、GSK-3β和β-catenin蛋白的表达。采用双荧光素酶报告基因实验验证miR-205-5p对RAP2B的靶向作用。结果与正常胃黏膜细胞相比,4种胃癌细胞中miR-205-5p的表达显著降低,RAP2B的表达显著升高(P<0.05)。过表达miR-205-5p或抑制RAP2B表达均可显著抑制AGS细胞的增殖、迁移和侵袭,抑制cyclin D1、MMP-2和MMP-9蛋白的表达(P<0.05)。miR-205-5p可负性调控RAP2B的表达。结论miR-205-5p通过靶向RAP2B抑制胃癌细胞的增殖、迁移和侵袭。Objective To investigate the effect of miR-205-5p on the proliferation,migration and invasion of gastric cancer cells by targeting at RAS oncogene family(RAP2B).Methods RT-qPCR and Western blot were used to detect the expression of miR-205-5p and RAP2B in gastric cancer cell lines AGS,MGC803,MKN-28,SGC-7901 and normal gastric mucosal cell line GES-1.Over expressing miR-205-5p,inhibition of RAP2B,cell viability was detected by MTT assay;cell migration and invasion ability were detected by Transwell method;The expression of RAP2B,cyclin D1,MMP-2,MMP-9 GSK-3βandβ-catenin proteins were detected by Western blot.The dual luciferase reporter gene assay was used to verify the targeting of miR-205-5p to RAP2B.ResultsCompared with normal gastric mucosal cells,the expression of miR-205-5p was significantly decreased in four gastric cancer cell lines,and the expression of RAP2B was significantly increased(P<0.05).The over-expression of miR-205-5p or inhibition of RAP2B inhibited the proliferation,migration and invasion of AGS cells and inhibited the expression of cyclin D1,MMP-2 and MMP-9 proteins(P<0.05).miR-205-5pnegatively regulatedthe expression of RAP2B.Conclusions miR-205-5p inhibits the proliferation,migration and invasion of gastric cancer cells in vitro by targeting at RAP2B.
关 键 词:miR-205-5p RAP2B 胃癌 增殖 迁移
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