miR-181b-5p靶向PTEN介导PI3K/Akt通路对弥漫大B细胞淋巴瘤增殖和侵袭的影响

Effect of miR-181b-5p Targeting PTEN Mediating PI3K/Akt Pathway on Proliferation and Invasion of Diffuse Large B-cell Lymphoma

目的探讨微小RNA(miR)-181b-5p靶向调控10号染色体上缺失的磷酸酶及张力蛋白同源基因(PTEN)对弥漫大B细胞淋巴瘤细胞增殖和侵袭的影响及其机制。方法第3代对数期SU-DHL-4细胞随机分为control组、mimic NC组、mimic组、inhibitor NC组和inhibitor组,qRT-PCR法检测各组miR-181b-5p和PTEN基因表达量,CCK-8法检测细胞增殖率,Transwell实验检测迁移和侵袭细胞数,双荧光素酶报告基因检测miR-181b-5p和PTEN之间的靶向关系,蛋白印迹法检测PTEN、磷酸化磷脂酰肌醇3-激酶(p-PI3K)和磷酸化蛋白激酶B(p-Akt)蛋白表达量。结果与mimic NC组比较,mimic组miR-181b-5p基因表达量以及p-PI3K和p-Akt蛋白表达量升高,PTEN基因和蛋白表达量降低,细胞增殖率及迁移和侵袭率升高(P<0.05);与inhibitor NC组比较,inhibitor组miR-181b-5p基因表达量以及p-PI3K和p-Akt蛋白表达量降低,PTEN基因和蛋白表达量升高,细胞增殖率及迁移和侵袭率降低(P<0.05)。从机制上看,miR-181b-5p靶向调控PTEN。结论下调miR-181b-5p可抑制弥漫大B细胞淋巴瘤细胞增殖、迁移和侵袭,其可能是通过靶向调控PTEN激活PI3K/Akt信号通路发挥作用。

Objective To investigate the effects of miR-181b-5p on the proliferation and invasion of diffuse large B-cell lymphoma cells by targeting phosphatase andtensin homologue deleted on chromosome 10(PTEN)and its mechanism.Methods The third generation of logarithmic SU-DHL-4 cells were randomly divided into the control group,mimic NC group,mimic group,inhibitor NC group and inhibitor group.The expression levels of miR-181b-5p and PTEN genes in each group were detected by qRT-PCR,and the cell proliferation rate was detected by CCK-8.The number of migrating and invasive cells was detected by Transwell assay,the targeting relationship between miR-181b-5p and PTEN was detected by dual luciferase reporter gene,and the protein expression levels of PTEN,phosphorylphosphatidylinositol 3-kinase(p-PI3K)and phosphorylated protein kinase B(p-Akt)were detected by western blot.Results Compared with mimic NC group,miR-181b-5p gene expression,cell proliferation rate,and p-PI3K and p-AKT protein expression were increased in mimic group,while PTEN gene and protein expression were decreased,and the number of migrating and invading cells were increased(P<0.05).Compared with inhibitor NC group,miR-181b-5p gene expression,cell proliferation rate,P-PI3K and P-Akt protein expression were decreased,PTEN gene and protein expression were increased,and the number of migrating and invading cells were decreased in inhibitor group(P<0.05).From the perspective of mechanism,miR-181b-5p targets to regulate PTEN.Conclusion Down-regulation of miR-181b-5p can inhibit proliferation,migration and invasion of diffuse large B-cell lymphoma cells,possibly through targeted regulation of PTEN to activate PI3K/Akt signaling pathway.