摘要
目的:探讨miR-139过表达对急性髓系白血病肿瘤坏死因子相关凋亡诱导配体(TNF-related apoptosis inducing ligand,TRAIL)耐受性的影响。方法:采用浓度为100、200、300和400 ng/ml TRAIL处理人急性髓系白血病K562、HL-60细胞及相对应的多药耐药K562/A02和HL-60/ADM细胞,采用CCK8法计算IC_(50)值,以IC_(50)法评价急性髓系白血病细胞对TRAIL的敏感性。RT-PCR检测miR-139在TRAIL敏感细胞和耐药细胞中的表达。采用脂质体2000将miR-139 mimics转染至TRAIL耐药细胞株,RT-PCR检测转染效果,CCK8法检测细胞活力及IC_(50)值,流式细胞仪检测细胞凋亡率。结果:在K562、K562/A02、HL-60和HL-60/ADM细胞中,HL-60/ADM细胞对TRAIL最为敏感,而K562细胞对TRAIL的耐药性最强。与敏感细胞株HL-60/ADM相比,miR-139在耐药细胞株K562中表达显著下降。转染miR-139 mimics后,K562细胞中miR-139表达和细胞凋亡率明显升高,而细胞存活率及IC_(50)值明显降低。结论:miR-139在TRAIL耐药细胞株K562中低表达,过表达miR-139可能通过促进TRAIL诱导的细胞凋亡降低K562细胞对TRAIL的耐受性。
Objective:To investigate the effect of overexpression of miR-139 on TNF-related apoptosis inducing ligand(TRAIL)tolerance in acute myeloid leukemia.Methods:K562,HL-60 cells and corresponding multidrug resistant K562/A02 and HL-60/ADM cells in human acute myeloid leukemia were treated with concentration of 100,200,300 and 400 ng/ml TRAIL.IC 50 value was calculated by CCK8 method.The sensitivity of tolerance in acute myeloid leukemia cells to TRAIL was evaluated by IC 50 method.The expression of miR-139 in TRAIL sensitive cells and drug-resistant cells was detected by RT-PCR.miR-139 mimics was transfected into TRAIL resistant strain by liposome 2000.The transfection effect was tested by RT-PCR.Cell viability and IC 50 value were detected by CCK8.The rate of apoptosis was detected by flow cytometry.Results:In K562,K562/A02,HL-60 and HL-60/ADM cells,HL-60/ADM cells were the most sensitive,while K562 cells had the strongest resistance to TRAIL.Compared with the sensitive cell line HL-60/ADM,miR-139 expression in the drug resistant cell line K562 was decreased significantly.After transfection of miR-139 mimics,the expression of miR-139 and apoptosis rate in K562 cells were increased significantly,while cell viability and IC 50 were decreased significantly.Conclusion:miR-139 is low in TRAIL resistant cell line K562.Overexpression of miR-139 may reduce the tolerance of K562 cells to TRAIL by promoting TRAIL induced apoptosis.
作者
姚金晓
杨如玉
马海龙
李超
魏旭东
Yao Jinxiao;Yang Ruyu;Ma Hailong;Li Chao;Wei Xudong(Department of Hematology,Nanyang City Center Hospital Affiliated to Zhengzhou University,Henan Nanyang 473000,China;;Department of Hematology,Cancer Hospital Affiliated to Zhengzhou University,Henan Zhengzhou 450008,China)
出处
《现代肿瘤医学》
CAS
2019年第1期9-12,共4页
Journal of Modern Oncology
基金
国家自然科学基金面上项目(编号:81170520)
