has-miR-150对Jurkat细胞增殖和凋亡的影响及其机制  

Effect of has-miR-150 on Proliferation and Apoptosis of Jurkat Cells and Its Mechanism

作　　者：王莹[1] 桑威[1] 孙财[1] 曾令宇[1] 徐开林[1] 

机构地区：[1]徐州医学院附属医院血液科,江苏徐州221002

出　　处：《中国实验血液学杂志》2015年第1期94-98,共5页Journal of Experimental Hematology

基　　金：江苏省自然科学基金(BK2012572);江苏省卫生厅科研基金(H201216)

摘　　要：目的:本研究旨在探讨has-miR-150对人急性T淋巴细胞白血病(T-ALL)细胞株Jurkat细胞增殖、凋亡的影响及其作用机制。方法:通过构建慢病毒载体过表达miR-150并感染Jurkat细胞,另设空白组和阴性对照组,应用实时定量PCR法检测miR-150的表达;应用CCK-8法和流式细胞术分别检测细胞增殖及凋亡情况,Western blot法检测细胞PI3K/Akt信号通路相关信号分子的蛋白表达。结果:构建的慢病毒载体过表达miR-150后感染Jurkat细胞,has-miR-150表达升高约2.06倍(P<0.05);与阴性对照相比,过表达miR-150后Jurkat细胞增殖能力明显下降(P<0.01),凋亡数明显增多(P<0.01),p-Akt及p-p65蛋白的表达明显下调,Akt蛋白的表达无明显变化。结论:过表达has-miR-150可以有效抑制Jurkat细胞的增殖并诱导其凋亡,对T-ALL可能具有潜在的抑癌作用,其机制可能与负调控PI3K/Akt/NF-κB信号通路有关。Objective： This study was to investigate the effect of has-miR-150 on the proliferation and apoptosis in human acute T lymphocytic leukemia （T-ALL） cell line Jurkat, and explore its mechanism. Methods： Lentivirus-has- miR-150 was constructed and transfected to Jurkat cells. The expression of miR-150 was detected by real time PCR; the cell proliferation and apoptosis were detected by CCK-8 method and Annexin V/7-AAD labeling, respectively; the cellrelated protein expressions of phosphatidylinositol-3-kinase （PI3K）/ serine! threonine kinase （Akt） signaling pathway were detected by Western blot. Results： The expression of miR-150 in infected Jurkat ceils was significantly upregulated by constructing lentivirus-has-miR-150. Compared to negative control （transfected with empty-vector lentivirus）, the cell proliferation after LV-miR-150 transfection was significantly inhibited and cell apoptosis was remarkably induced. Phosphorylation levels of P13K/Akt/NF-kB signaling pathway protein p-AM and p-p65 decreased, whereas no obvious change was found in the expression of Akt. Conclusion： miR-150 may be a putative oncoprotein in T-ALL cells. Overexpression of miR-150 has noticeable effects on the proliferation inhibition and apoptosis induction of Jurkat cells, which may be mediated by the negative regulation of PI3K/Akt/NF-KB signaling pathway.

关 键 词：has-miR-150 JURKAT细胞 细胞增殖 细胞凋亡 PI3K/Akt/NF-κB信号通路 

分 类 号：R733.71[医药卫生—肿瘤]