中国肿瘤生物治疗杂志2017，Vol.24Issue(10)：1063-1069,7.DOI:10.3872/j.issn.1007-385x.2017.10.004
沉默pim-3基因对黑色素瘤细胞B16的抑制作用
Inhibitory effect of target silencing Pim-3 gene on melanoma B16 cell line
摘要
Abstract
Objective:To observe the inhibitory effect of target silencing pim-3 on mouse melanoma B16 cell line.Methods:Pim-3-shRNA vector andpim-3 over-expressing vector (pim-3-MIGR1) were respectively transfected into B16 melanoma cells by liposomc,and the changes in mRNA and protein expression ofpim-3 was measured by qPCR and Western blotting assay,respectively;the apoptosis of B16 cells was detected by AnnexinV/PI staining and TUNEL staining;the expressions of apoptosis-related factors,such as p-Bad,Bcl-2,Bcl-xl,Bax,caspase-3,were evaluated by qPCR and Western blotting;MTT assay was applied to confirm the proliferation of B16 cells;Flow cytometry was used to detect cell cycle;RTCAxCELLigence system was performed to confirm cell migration;and Transwell assay was applied to examine cell invasion.Results:pim-3-shRNA significantly reduced while pim-3-MIGR1 significantly enhanced pim-3 expression in B 16 cells at both mRNA and protein levels (all P＜0.05).Pim-3-shRNA also effectively induced the apoptosis of B16 cells;however,co-transfection with pim-3-MIGR1 obviously reversed the apoptosis induced by Pim-3 silencing (P＜0.05).Further studies found pim-3-shRNA significantly downregulated the expressions of apoptosis-related molecules (p-Bad,Bcl-2 and Bcl-xl),but remarkably up-regulated Bax expression,and finally resulted in increased caspase-3 activity (all P＜0.05).The proliferation and migration of B 16 cells were significantly inhibited by pim-3 silencing (all P＜0.05);however,there was no obvious change in cell cycle.Conclusion:Pim-3-specific gene silencing effectively promoted the apoptosis and inhibited both the proliferation and migration of B 16 cells,suggesting that pim-3 may serve as a potential effective gene therapy target for melanoma.关键词
黑色素瘤/pim-3基因/B16细胞/凋亡/增殖/侵袭Key words
melanoma/pim-3 gene/B 16 cell/apoptosis/proliferation/invasion分类
医药卫生引用本文复制引用
刘静,于馨,胡渊,张彩..沉默pim-3基因对黑色素瘤细胞B16的抑制作用[J].中国肿瘤生物治疗杂志,2017,24(10):1063-1069,7.基金项目
国家自然科学基金资助项目(No.81472646,No.81273220,No.91442114).Project supported by the National Natural Science Foundation of China(No.81472646,No.81273220,No.91442114) （No.81472646,No.81273220,No.91442114）
