miR-876靶向Pax6抑制胃癌细胞增殖、侵袭和迁移

miR-876 targeting Pax6 inhibits the proliferation,invasion and migration of gastric cancer cells

目的探讨miR-876靶向Pax6对胃癌细胞增殖、侵袭和迁移的影响。方法采用RT-qPCR检测胃癌组织与癌旁组织中miR-876和Pax6的表达水平,分析miR-876表达与胃癌患者临床指标的关系。检测胃上皮细胞GES-1以及胃癌细胞MGC-803、BGC-823、SGC-7901中miR-876的表达水平。将MGC-803细胞分为对照组、miR-876 mimic组、mimic-NC组和miR-876 mimic+pcDNA-3.1-Pax6组,采用RT-qPCR和Western blotting检测各组细胞中miR-876以及Pax6的mRNA和蛋白表达水平;MTT检测各组细胞增殖情况;Transwell检测各组细胞迁移和侵袭情况;双荧光素酶实验验证miR-876与Pax6的靶向关系。结果与癌旁组织相比,miR-876在胃癌组织中低表达(t=13.962,P<0.05),而Pax6 mRNA和蛋白在胃癌组织中均呈高表达(t=23.368,13.757;P<0.05),且两者呈负相关(r=-0.434,P<0.05)。miR-876低表达与胃癌患者TNM分期(χ^(2)=12.000,P<0.05)和淋巴结转移(χ^(2)=15.705,P<0.05)有关。细胞实验发现,miR-876在胃癌细胞系MGC-803、BGC-823、SGC-7901中均低表达(P<0.05);转染miR-876 mimic后,miR-876表达水平显著升高,细胞增殖能力减弱,迁移和侵袭能力均受到抑制(P<0.05)。双荧光素酶实验结果表明:miR-876能够靶向结合Pax6。与mimic-NC组比,miR-876 mimic组Pax6mRNA和蛋白表达水平显著下降(P<0.05);与miR-876 mimic组比,miR-876 mimic+pcDNA-3.1-Pax6组细胞增殖、迁移和侵袭能力均显著提高(P<0.05)。结论miR-876靶向Pax6可抑制胃癌细胞MGC-803的增殖、侵袭和迁移。

Objective To investigate the effects of miR-876 targeting Pax6 on the proliferation,invasion and migration of gastric cancer cells.Methods RT-qPCR was used to detect the expressions of miR-876 and Pax6 in gastric cancer tissues and para-carcinoma tissues,and the relationship was analyzed between miR-876 expression and clinical indicators of gastric cancer patients.The expression of miR-876 in gastric epithelial cells GES-1 and gastric cancer cells MGC-803,BGC-823 and SGC-7901 were also detected.MGC-803 cells were divided into control group,miR-876 mimic group,mimic NC group and miR-876 mimic+pcDNA-3.1-Pax6 group.The expressions of miR-876 as well as Pax6 mRNA and protein in each group were detected by RT-qPCR and Western blotting.The cell proliferation in each group was detected by MTT,and the cell migration and invasion by Transwell.The targeting relationship between miR-876 and Pax6 was verified by double luciferase assay.Results Compared with para-carcinoma tissues,the miR-876 was lowly expressed in gastric cancer tissues(t=13.962,P<0.05),while Pax6 mRNA and protein were highly expressed in gastric cancer tissues(t=23.368,13.757,all P<0.05),and there was a negative correlation between them(r=-0.434,P<0.05).The low expression of miR-876 was associated with TNM stage(χ^(2)=12.000,P<0.05)and lymph node metastasis(χ^(2)=15.705,P<0.05).Cell experiment found that miR-876 was also lowly expressed in gastric cancer cell line MGC-803,BGC-823,SGC-7901(P<0.05).After miR-876 mimic transfection,miR-876 expression levels increased significantly,and the cell proliferation was weakened,both migration and invasion were inhibited(P<0.05).The results of double luciferase assay showed that miR-876could target Pax6.Compared with the mimic-NC group,Pax6 mRNA and protein expression levels decreased significantly in the miR-876 mimic group(P<0.05).Compared with the miR-876 mimic group,the proliferation,migration and invasion ability of miR-876 mimic+pcDNA-3.1-Pax6 group were significantly improved(P<0.05).Conclusion miR-876 targeting Pax6 inhibits the proliferation,invasion and migration MGC-803 gastric cancer cells.