miR-15b-5p靶向HDGF抑制胃癌细胞增殖并促进凋亡

目的:研究miR-15b-5p和HDGF对胃癌细胞增殖和凋亡的影响，并探讨其作用机制。方法:采用qRT-PCR法检测胃癌细胞中miR-15b-5p的表达；通过软件预测miR-15b-5p的下游靶基因；将对数生长期AGS细胞随机分为miR-15b-5p组（转染miR-15b-5p模拟物）、miR-NC组(转染阴性对照)、Scramled 组(转染阴性对照)、shHDGF组(转染质粒pcDNA3.1-shHDGF)、Vector+miR-15b-5p组(miR-15b-5p模拟物和pcDNA3.1载体共转染)、HDGF+miR-15b-5p组(miR-15b-5p模拟物和HDGF共转染)，均以脂质体法转染；MTT法和流式细胞术检测各组细胞的增殖和凋亡变化；Western blot法和双荧光素酶报告基因实验分析miR-15b-5p对HDGF的靶向调控作用。结果:与正常胃黏膜上皮细胞GES-1相比，胃癌细胞SGC-7901和AGS中miR-15b-5p低表达，且过表达miR-15b-5p可抑制AGS细胞的增殖并促进凋亡。HDGF是miR-15b-5p的潜在靶标，miR-15b-5p能够抑制HDGF表达。而回补HDGF可逆转miR-15b-5p抑制胃癌细胞增殖与促进凋亡的作用。结论:miR-15b-5p可抑制胃癌细胞的增殖并促进凋亡，其作用机制可能与靶向抑制HDGF有关，提示miR-15b-5p可能成为胃癌诊断与治疗的潜在靶点。

miR-15b-5p inhibits cell proliferation and promotes apoptosis of gastric cancer by targeting HDGF

Objective:To study the effect of miR-15b-5p and HDGF on the proliferation and apoptosis of gastric cancer cells,and explore its mechanism.Methods:The expression of miR-15b-5p in gastric cancer cells was detected by qRT-PCR.The downstream target genes of miR-15b-5p were predicted by software.Logarithmic growth phase AGS cells were randomly divided into miR-15b-5p group (transfected miR-15b-5p),miR-NC group (transfected negative control),Scramled group (transfected negative control),shHDGF group (transfected plasmid pcDNA3.1-shHDGF),Vector+miR-15b-5p group (miR-15b-5p and empty carrier co-transfection),HDGF+miR-15b-5p group (miR-15b-5p and HDGF co-transfection),transfected by liposome method.Cell proliferation and apoptosis of each group cells were detected by MTT assay and flow cytometry.Western blot and dual luciferase reporter gene assay were used to analyze the targeting regulation of miR-15b-5p on HDGF.Results:Compared with GES-1,a normal gastric mucosal epithelial cell,miR-15b-5p was been down-regulated in gastric cancer cells SGC-7901 and AGS,and overexpression of miR-15b-5p decreased cell proliferation and induced apoptosis.HDGF was a underlying target of miR-15b-5p,and miR-15b-5p could decrease the expression of HDGF.Replenishing HDGF could rescue the effect of miR-15b-5p on proliferation and apoptosis of AGS cells.Conclusion:miR-15b-5p could inhibit cell proliferation and induce apoptosis in gastric cancer cells and its mechanism may be related to targeting HDGF.It suggests that miR-15b-5p may be a potential target for the diagnosis and treatment of gastric cancer.