慢病毒载体上调GADD45α增强伊马替尼耐药株K562IR细胞的药物敏感性  

Upregulation of GADD45αby Lentivirus vector enhances the drugsensitivity of imatinib-resistant K562IR cells

作　　者：刘雪妮 王超[1] 程艳红 杨鹏[1] Liu Xueni;Wang Chao;Cheng Yanhong(Dept of Blood Transfusion, The First Affiliated Hospital of Anhui Medical University, Hefei 230022)

机构地区：[1]安徽医科大学第一附属医院输血科,合肥230022

出　　处：《安徽医科大学学报》2021年第12期1920-1924,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:1808085MH273)。

摘　　要：目的探讨通过慢病毒载体上调GADD45α基因对伊马替尼(IM)耐药株K562IR细胞的增殖、迁移及凋亡的影响。方法培养K562细胞及伊马替尼耐药细胞K562IR,采用实时荧光定量PCR技术(qRT-PCR)和Western blot法检测K562和K562IR细胞中GADD45αmRNA和蛋白的表达水平;采用过表达GADD45α的慢病毒载体及对照载体转染于K562IR细胞中,分别为Lv-GADD45α和Lv-NC,通过Western blot检测上调后各组细胞GADD45α蛋白表达水平,并进一步用CCK-8法、Tanswell法和Annexin V-FITC/PI双染法检测转染后细胞增殖、迁移和凋亡的变化。结果K562IR细胞中的GADD45α蛋白和mRNA表达水平低于K562细胞,GADD45α基因转染后,Lv-GADD45α组细胞中的GADD45α蛋白表达水平高于Lv-NC组。Lv-GADD45α组细胞在伊马替尼处理后,IC_(50)值低于Lv-NC组,凋亡率高于Lv-NC组,同时细胞迁移能力也下降,差异有统计学意义(P<0.05)。结论通过上调伊马替尼耐药株K562IR细胞中GADD45α基因,可以增强慢性髓系白血病细胞K562IR对于伊马替尼的敏感性,抑制白血病细胞的增殖、促进其凋亡。Objective To investigate the effects of up-regulation of GADD45αby Lentivirus vector on the proliferation,migration and apoptosis of imatinib-resistant K562IR cells.Methods K562 cells and imatinib-resistant K562IR cells were cultured.The expression levels of GADD45αmRNA and protein in K562 and K562IR cells were detected by qRT-PCR and Western blot.The Lentivirus vector that over-expressed GADD45αand control were transfected in K562IR cells,respectively Lv-GADD45αand Lv-NC.The expression level of GADD45αin each group was detected by Western blot,and the changes in cell proliferation,migration and apoptosis after transfection were further detected by CCK-8,Tanswell and Annexin V-FITC/PI double staining.Results The expression levels of GADD45αprotein and mRNA in K562IR cells were lower than those in K562 cells.After GADD45αwas transfected,the expression levels of GADD45αin Lv-GADD45αgroup were higher than those in Lv-NC group.After imatinib treatment,the IC_(50) value of Lv-GADD45αgroup was lower than that of Lv-NC group,the apoptosis rate of cells in Lv-GADD45αgroup was higher than that of the Lv-NC group,and the cell migration ability also decreased,with statistically significant difference(P<0.05).Conclusion By up-regulating GADD45αin imatinib-resistant K562IR cells,the sensitivity of K562IR to imatinib in chronic myeloid leukemia cells is enhanced,leading to proliferation inhibition and the apoptosis of chronic myeloid leukemia cells.

关 键 词：慢性粒细胞白血病 K562 GADD45α 伊马替尼 耐药性 

分 类 号：R733.7[医药卫生—肿瘤]