miR-452经Hedgehog信号通路抑制胃癌HGC-27细胞增殖和迁移的研究

miR-452 in inhibition on proliferation and migration of gastric cancer HGC-27 cells through Hedgehog signaling pathway

目的研究微小核糖核酸452(miR-452)表达对胃癌HGC-27细胞增殖和迁移的影响及机制。方法收集2017年7月至2018年12月收治的胃癌患者手术切除的胃癌组织及对应的癌旁组织各50例,Real-timePCR方法测定胃癌患者胃癌、癌旁组织和外购的胃癌、正常胃黏膜细胞中miR-452表达变化。在胃癌HGC-27细胞中转染miR-452mimics;噻唑蓝(MTT)测定细胞增殖;Transwell小室测定细胞侵袭和迁移;Westernblot测定Hedgehog信号通路关键蛋白[音猬因子(Shh)、胶质瘤相关癌基因(Gli)1]和肿瘤侵袭转移相关蛋白[基质金属蛋白酶(MMP)-2和MMP-9]的表达。观察Hedgehog信号通路激活剂PM处理后胃癌HGC-27细胞miR-452表达水平的变化及其对细胞增殖、侵袭和迁移能力的影响。结果胃癌患者胃癌组织中miR-452表达水平低于癌旁组织(P<0.05),外购的胃癌细胞中miR-452表达水平低于正常胃黏膜细胞(P<0.05)。转染miR-452mimics后的胃癌HGC-27细胞中miR-452表达水平升高,细胞增殖、侵袭和迁移能力均降低,细胞中Shh、Gli1、MMP-2和MMP-9蛋白表达水平均下降(P均<0.05)。Hedgehog信号通路激活剂PM处理后胃癌HGC-27细胞miR-452表达上调,其增殖、侵袭和迁移能力均升高(P均<0.05)。结论miR-452在胃癌中低表达,而上调其表达可调控Hedgehog信号通路,降低胃癌细胞增殖和迁移能力。

Objective To investigate the effect and mechanism of microRNA-452(miR-452) expression on the proliferation and migration of gastric cancer cell HGC-27.Methods A total of 50 gastric patients who received operation at Chengdu Seventh People′s Hospital from July 2017 to December 2018 were selected,and their resected gastric cancer tissues and corresponding adjacent tissues were collected.Real-time PCR was used to detect the expression of miR-452 in gastric cancer,adjacent tissues,purchased gastric cancer and normal gastric mucosa cells.miR-452 mimics were transfected into HGC-27 cells;Cell proliferation was measured by MTT;invasion and migration were measured by Transwell chamber.Western blot was used to detect the expression of key proteins in Hedgehog signaling pathway[sonic hedgehog(Shh),glioma-related oncogene(Gli)1] and tumor invasion and metastasis-related proteins[matrix metalloproteinase(MMP)-2 and MMP-9].After deal with the Hedgehog signaling pathway activator PM,the expression changes of miR-452 in gastric cancer HGC-27 cells and its effect on cell proliferation,invasion and migration were observed.Results The expression level of miR-452 in gastric cancer tissues from patients was lower than that in adjacent tissues(P<0.05),and the expression level of miR-452 in parchased gastric cancer cells was lower than that in normal gastric mucosa cells(P<0.05).After transfection of miR-452 mimics,the expression level of miR-452 in HGC-27 cells was increased,the proliferation,invasion and migration ability of cells were decreased,and the protein expression levels of Shh,Gli1,MMP-2 and MMP-9 were decreased(all P<0.05).The expression of miR-452 was up-regulated in HGC-27 cells treated with Hedgehog signaling pathway activator PM,and its proliferation,invasion and migration abilities were increased(all P<0.05).Conclusion The expression of miR-452 is low in gastric cancer,while the up-regulation of miR-452 expression can regulate Hedgehog signaling pathway and reduce the proliferation and migration of gastric cancer cells.