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

摘要:

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（χ²=12.000， P<0.05）和淋巴结转移（χ²=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Pax6 mRNA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。

Abstract:

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 (χ²=12.000, P<0.05) and lymph node metastasis (χ²=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-876 could 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.