沉默miR-4320抑制胃癌细胞增殖和迁移的作用机制研究

Mechanism of silencing miR-4320 expression in inhibiting proliferation and migration of gastric cancer cells

目的探讨微小RNA(microRNA)-4320对胃癌MGC803细胞增殖、迁移的影响及机制。方法分别采用实时定量聚合酶链式反应(qRT-PCR)检测miR-4320在4种胃癌细胞株(MGC803、HS-746T、SGC7901、BGC823)中的表达情况。将携带miR-4320干扰片段的重组慢病毒和空白慢病毒感染MGC803细胞,设为si-miR-4320组和NC组。噻唑蓝比色法和Transwell小盒实验分别检测下调miR-4320后MGC803细胞增殖和迁移情况。生物信息学软件RNAhybrid预测miR-4320目的基因。通过双荧光素酶报告基因实验验证miR-4320与目的基因的靶向关系。分别采用qRT-PCR和Western blot检测miR-4320目的基因的表达。计量数据以均数±标准差(±s)表示,组间比较应用t检验或单因素方差分析。结果4种胃癌细胞株miR-4320的表达均显著高于正常胃黏膜上皮细胞(P<0.01)。NC组和si-miR-4320组MGC803细胞中miR-4320表达分别为8.19±1.00和1.09±0.31,miR-4320干扰片段显著降低miR-4320的表达(P<0.01)。si-miR-4320组MGC803细胞的吸光度明显低于NC组(P<0.05),迁移能力明显低于NC组(P<0.01)。细胞因子信号抑制因子(SOCSI)是miR-4320的目的基因(P<0.01)。与NC组相比,si-miR-4320组SOCS1基因表达明显上调(P<0.01)。结论miR-4320在胃癌细胞株中表达升高,下调miR-4320的表达能够通过诱导SOCS1基因表达,抑制胃癌MGC803细胞的增殖及迁移能力。

Objective To explore the effect and mechanism of microRNA(miRNA)-4320 on the proliferation and migration of gastric cancer MGC803 cells.Methods Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-4320 in four gastric cancer cell lines(MGC803,HS-746T,SGC7901,BGC823)MGC803 cells were infected with recombinant lentivirus carrying miR-4320 interference fragments or blank lentivirus,and set as si-miR-4320 group and NC group.Thiazole blue colorimetry and Transwell small box experiment were used to detect the proliferation and migration of MGC803 cells after miR-4320 was down-regulated.The bioinformatics software RNAhybrid was used to predict the target gene of miR-4320.The targeting relationship between miR-4320 and target gene was verified by dual-luciferase reporter gene experiment.qRT-PCR and Western blot were used to detect the expression of miR-4320 target gene.Measurement data were expressed as mean±standard deviation(±s),and t-test or one-way ANOVA was used for comparison between groups.Results The expression of miR-4320 in the four gastric cancer cell lines was significantly higher than that of normal gastric mucosal epithelial cells(P<0.01).The expression of miR-4320 in MGC803 cells in the NC group and the si-miR-4320 group were 8.19±1.00 and 1.09±0.31,respectively.The miR-4320 interference fragment significantly reduced the expression of miR-4320(P<0.01).The absorbance of MGC803 cells in the si-miR-4320 group was significantly lower than that of the NC group(P<0.05),and the migration ability was significantly lower than that of the NC group(P<0.01).Suppressor of cytokine signaling1(SOCSI)is the target gene of miR-4320.Compared with the NC group,the SOCS1 gene expression in the si-miR-4320 group was significantly up-regulated(P<0.01).Conclusions The expression of miR-4320 is increased in gastric cancer cell lines.Down-regulating the expression of miR-4320 can inhibit the proliferation and migration of gastric cancer MGC803 cells by inducing the expression of SOCS1 gene.