miR-6751-3p在胃癌细胞增殖和迁移中的作用机制研究

Study on the mechanism of miR-6751-3p affecting the proliferation and migration of gastric cancer cells

目的:探讨微小RNA(microRNA,miRNA)-6751-3p表达对胃癌细胞增殖和迁移的影响及其分子机制。方法:采用实时定量聚合酶链式反应(qRT-PCR)检测胃癌细胞株(MGC803、BGC823、SGC7901、HS-746T)和正常胃黏膜上皮细胞(GES-1)中miR-6751-3p的表达水平。将miR-6751-3p表达水平最低的胃癌细胞株分为对照组和实验组,分别转染miR-NC和miR-6751-3p模拟物。qRT-PCR检测两组细胞miR-6751-3p的表达水平。分别采用CCK-8法和划痕愈合实验检测miR-6751-3p过表达细胞的增殖和迁移情况。通过Deepbase v2.0和microRNA.org在线网站预测miR-6751-3p的潜在靶基因,采用双荧光素酶报告基因实验进行验证。qRT-PCR和Western blot检测miR-6751-3p过表达细胞靶基因的表达。结果:与正常胃黏膜上皮细胞相比,miR-6751-3p在胃癌细胞株中表达明显下调(P<0.05),表达水平最低的细胞株是MGC803细胞(P<0.01)。与对照组相比,miR-6751-3p过表达可抑制MGC803细胞的增殖能力(P<0.05)。对照组和实验组MGC803细胞划痕愈合率分别为(65.14±5.65)%和(23.40±6.78)%。与对照组相比,实验组MGC803细胞划痕愈合率显著降低(t=4.73,P<0.001)。在线网站预测miR-6751-3p的靶基因可能是脂肪酸结合蛋白5(FABP5),miR-6751-3p可互补结合FABP5信使RNA(mRNA)(P<0.01)。与对照组相比,miR-6751-3p过表达可抑制MGC803细胞中FABP5基因的表达(t=4.01,P<0.01)。结论:胃癌细胞株中miR-6751-3p呈低表达,miR-6751-3p过表达可通过下调FABP5基因能抑制胃癌MGC803细胞增殖和迁移能力。

Objective:To explore the influence of microRNA(miRNA)-6751-3p expression on the proliferation and migration of gastric cancer cells and its molecular mechanism.Methods:Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression level of miR-6751-3p in gastric cancer cell lines(MGC803,BGC823,SGC7901,HS-746T)and normal gastric mucosal epithelial cells(GES-1).The gastric cancer cell lines with the lowest expression level of miR-6751-3p were divided into control group and experimental group,and were transfected with miR-NC and miR-6751-3p mimics respectively.qRT-PCR detected the expression level of miR-6751-3p in the two groups of cells.CCK-8 method and scratch healing test were used to detect the proliferation and migration of miR-6751-3p overexpressing cells.The potential target genes of miR-6751-3p were predicted through Deepbase v2.0 and microRNA.org online websites,and the dual luciferase reporter gene experiment was used to verify.qRT-PCR and Western blot were used to detect the expression of target genes in miR-6751-3p overexpression cells.Results:Compared with normal gastric mucosal epithelial cells,the expression of miR-6751-3p was significantly down-regulated in gastric cancer cell lines(P<0.05),and the cell line with the lowest expression level was MGC803 cells(P<0.01).Compared with the control group,overexpression of miR-6751-3p can inhibit the proliferation ability(P<0.05).The scratch healing rate of MGC803 cells in the control group and the experimental group were(65.14±5.65)%and(23.40±6.78)%,respectively.Compared with the control group,the scratch healing rate of MGC803 cells in the experimental group was significantly lower(t=4.73,P<0.01).The online website predicts that the target gene of miR-6751-3p may be fatty acid binding protein 5(FABP5),and miR-6751-3p can complementally bind FABP5 messenger RNA(mRNA)(t=4.01,P<0.01).Compared with the control group,overexpression of miR-6751-3p can inhibit the expression of FABP5 gene in MGC803 cells(P<0.01).Conclusion:The expression of miR-6751-3p in gastric cancer cell lines is low,and the overexpression of miR-6751-3p can inhibit the proliferation and migration of gastric cancer MGC803 cells by down-regulating the FABP5 gene.