摘要
目的:通过沉默人多发性骨髓瘤细胞株RPM18226中miRNA-181a的表达,观察RPM18226细胞的增殖、迁移和细胞周期的变化,并探讨其可能的作用机制。方法:实时荧光定量PCR检测多发性骨髓瘤患者和健康者血清标本中miRNA-181a的表达水平;转染miRNA-181a inhibitor后,CCK-8法与集落形成实验检测细胞的增殖能力,划痕实验检测细胞的迁移能力,流式细胞术检测细胞的周期变化,Western blot法检测cyclin D1、p-PI3K和pAkt蛋白水平的变化。结果:多发性骨髓瘤患者血清中miRNA-181a呈高表达,显著高于正常人;转染miRNA-181a inhibitor后,RPM18226细胞的存活率和集落形成能力下降,迁移能力降低,G_0/G_1期细胞比例明显减少,S期细胞比例增多,cyclin D1蛋白表达显著低于正常对照组,PI3K和Akt的磷酸化水平明显低于正常对照组。结论:沉默miRNA-181a的表达能够抑制RPM18226细胞的增殖,并降低细胞的迁移能力,可能与细胞周期及PI3K/Akt信号通路有关。
AIM：To investigate the effect of miRNA-181 a inhibition on the proliferation,migration and cell cycle of the human multiple myeloma cell line RPMI8226.METHODS：Real-time PCR was used to detect miRNA481 a expression in serum samples from multiple myeloma or healthy subjects.After transfection with miRNA-181 a inhibitor,the cell viability was examined by CCK-8 assay and colony formation assay.The cell migration ability was analyzed by wound healing assay.The cell cycle was detected by flow cytometry.Moreover,the protein level of cyclin D1 and the phosphorylation of PI3 K and Akt were determined by Western blot.RESULTS：The expression of miRNA-181 a was significantly increased in the serum from multiple myeloma patients as compared with healthy group.Inhibition of miRNA-181 a expression by transfection with miRNA-181 a inhibitor remarkably decreased the cell viability,migratory ability,the population of G0/G1 phase and cyclin Dl protein expression in the RPMI8226 cells.However,the population of S phase and the phosphorylation of PI3 K and Akt were reduced.CONCLUSION：Down-regulation of miRNA-181 a inhibits the viability and migratory ability in the RPMI8226 cells via inhibition of cell cycle and PI3K/Akt signaling pathway.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2017年第1期33-37,共5页
Chinese Journal of Pathophysiology
基金
宁波市自然科学基金资助项目(No.2012A610202)
