MicroRNA-17-92 簇对 K562 细胞生物学特性的影响及机制初探简 |
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引用本文: | 杨爽,孙慧燕,肖凤君,李庆芳,许俊,郭子宽,王恒湘,王立生.MicroRNA-17-92 簇对 K562 细胞生物学特性的影响及机制初探简[J].中国实验血液学杂志,2014(1):20-24. |
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作者姓名: | 杨爽 孙慧燕 肖凤君 李庆芳 许俊 郭子宽 王恒湘 王立生 |
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作者单位: | [1]大连医科大学血液科,辽宁大连116044 [2]军事医学科学院放射与辐射医学研究所实验血液学研究室,北京100850 [3]空军总医院血液科,北京100142 [4]中国人民解放军总医院肿瘤中心,北京100853 |
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基金项目: | 国家自然科学基金(81170494; 81170460) |
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摘 要: | 本研究旨在探讨microRNA-17-92对K562细胞生物学特性的影响.以K562细胞系为模型,采用lipofectin2000转染miR-17-92 mimic,应用Q-PCR方法鉴定基因表达水平,CCK-8细胞增殖检测试剂盒检测K562细胞增殖活性,Annexin V-FITC/PI标记检测K562细胞的凋亡;细胞经过乙醇固定后利用流式细胞仪检测K562细胞周期变化,Western blotting检测相关蛋白Crk的表达.结果显示:K562细胞系转染miR-17-92 mimic后miR-17-92表达增高;miR-17-92高表达后K562细胞的增殖能力增强、在细胞周期方面由G1期向S期细胞增加;同时,miR-17-92 mimic能够抑制乏血清诱导的K562细胞凋亡;Western blot检测显示miR-17-92 mimic促进Crk蛋白的表达.结论:miR-17-92可以促进K562细胞增殖,抑制其凋亡并调控细胞周期变化.
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关 键 词: | miR-17-92簇 慢性髓性白血病 K562细胞系 |
Effect of MicroRNA-17-92 Cluster on the Biological Characteristics of K562 Cells and Its Mechanisms |
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Institution: | YANG Shuang;SUN Hui-Yan;XIAO Feng-Jun;LI Qing-Fang;XU Jun;GUO Zi-Kuan;WANG Heng-Xiang(Department of Hematology, Dalian Medical University, Dalian 116044, Liaoning Province, China;Department of Hematology,General Hospital of Chinese Air Force, Beijing 1000142, China;Department of Experimental Hematology, Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China;Cancer Center, Chinese PLA General Hospital, Beijing 100853, China) |
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Abstract: | The objective of this study was to explore the effects of microRNA-17-92 on the biological characteristics of K562 cells.The expression of miR-17-92 in K562 cells transfected with miRNA-17-92 mimic was detected by real time PCR.The effect of microRNA-17-92 on K562 cell proliferation was detected by CCK-8 method.Apoptosis of K562 cells was detected by Annexin V-PI labeling.Cell cycle distribution was determined by using flow cytometry.Western blot was performed to determine the protein levels of Crk.The results indicated that the transfection with miR-17-92 mimic increased expression of mature miR-17-92 in K562 cells.Compared with control group,cell proliferation and cell amount in S-phase of miR-17-92 mimic transfected group significantly increased,cell apoptosis decreased.The expression of signal connector protein Crk was greatly up-regulated in miR-17-92-mimic-transfected K562 cells.It is concluded that miR-17-92 can promote proliferation,inhibit apoptosis and regulate the cell cycle of K562 cells. |
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Keywords: | miR-17-92 cluster chronic myeloid leukemia K562 cell line |
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