miR-32通过调控PI3K/Akt信号通路影响骨肉瘤细胞的增殖

摘要

目的 探讨 microRNA-32(miR-32)在骨肉瘤细胞增殖中的作用及机制.方法 应用逆转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)技术检测骨肉瘤组织与正常组织及不同细胞株中miR-32的表达情况.采用CCK-8方法检测抑制 miR-32表达对人成骨肉瘤 MG-63细胞株增殖速率的影响.用荧光素酶实验检测 miR-32与人第10号染色体缺失的磷酸酶及张力蛋白同源基因(phosphatase and tensin homolog deleted on chromosome ten,PTEN)的3'-UTR特异性结合,采用 RT-PCR和蛋白质免疫印迹技术分别从 RNA 及蛋白质水平上验证 miR-32与PTEN的关系.结果 miR-32在骨肉瘤组织及细胞中通过与 PTEN 的3'-UTR靶向结合后激活了磷脂酰肌醇-3-激酶/丝氛酸-苏氨酸蛋白激酶(phosphatidylinositol-3-kinase/serine-threonine kinase,PI3K/Akt)信号通路,进而影响人成骨肉瘤 MG-63细胞的增殖.结论 miR-32通过 PI3K/Akt信号通路调控骨肉瘤细胞的增殖.%Objective To find out the function and mechanism of microRNA-32(miR-32)in osteosarcoma cell proliferation.Methods Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of miR-32 in osteosarcoma tissues,normal tissues and different cell lines.The effect of inhibiting miR-32 expression on the proliferation rate of human osteosarcoma MG-63 cell line was detected by CCK-8.Luciferase assay showed that miR-32 could bind specifically to the 3'-UTR of phosphatase and tensin homolog deleted on chromosome ten (PTEN)gene.At the same time,RT-PCR and western blotting were used to verify the relationship between miR-32 and PTEN from RNA and protein level respectively.Results miR-32 in osteosarcoma tissues and cells activate PI3K/Akt signaling pathway after binding to PTEN's 3'-UTR,which further influences the proliferation of MG-63 cells.Conclusion miR-32 regulates the proliferation of osteosarcoma cells through PI3K/Akt signaling pathway.