STAT3介导耐药白血病细胞抗凋亡和免疫逃逸机制的研究  

Mechanism of Anti Apoptosis and Immune Evasion in Drug-Resistant Leukemia Cells Mediated by STAT3

作　　者：贾祝霞 卢绪章 何金媛 蔡晓辉 秦伟 韩文敏 周民 徐卫[3] JIA Zhu-Xia;LU Xu-Zhang;HE Jin-Yuan;CAI Xiao-Hui;QIN Wei;HAN Wen-Min;ZHOU Min;XU Wei(Department of Hematology,The Affiliated Hospital of Nanjing Medical University,Changzhou No.2 People's Hospital,Changzhou 213000,China;Changzhou No.3 People's Hospital,Changzhou 213000,Jiangsu Province,China;Department of Hematology,The First Affiliated Hospital of Nanjing Medical University(Jiangsu Province Hospital),Nanjing 210029,China)

机构地区：[1]南京医科大学附属常州第二人民医院血液科,江苏常州213000 [2]常州第三人民医院,江苏常州213000 [3]南京医科大学第一附属医院(江苏省人民医院)血液科,江苏南京210029

出　　处：《中国实验血液学杂志》2020年第6期1796-1803,共8页Journal of Experimental Hematology

基　　金：常州市卫生计生委青年人才科技项目(QN201715),常州市高层次卫生人才培养工程(2016CZLJ027)。

摘　　要：目的:研究STAT3在耐药白血病细胞凋亡及免疫逃逸中的作用,并探讨白血病微小残留病引起的白血病复发的可能机制。方法:用pcDNA3.1-STAT3质粒转染K562细胞,建立耐药白血病细胞株K562/STAT3细胞。RQ-PCR和(或)Western blot检测空载质粒转染的对照细胞株K562/-细胞和K562/STAT3细胞中STAT3、BAX以及NKG2D配体的表达。流式细胞术检测K562/-细胞和K562/STAT3细胞的凋亡情况及NK细胞对K562/-细胞和K562/STAT3细胞的杀伤作用。结果:K562/STAT3细胞中总STAT3和STAT3磷酸化水平均明显升高,而且其耐药蛋白P-gp mRNA的表达明显升高(P<0.005)。与K562/-细胞相比,K562/STAT3细胞促凋亡基因BAX mRNA的表达水平明显下降(P=0.005),且多柔比星引起的K562/STAT3细胞凋亡明显减少(P=0.002);用STAT3抑制剂(SH-4-54)处理K562/STAT3细胞后,促凋亡基因BAX mRNA的表达水平明显升高(P=0.017),同时细胞凋亡也明显增加(P=0.005)。与K562/-细胞相比,K562/STAT3细胞NKG2D配体(MICA和ULBP1)mRNA的表达水平明显下降(P<0.0001),MICA和ULBP1蛋白的表达水平亦明显下降(MICA:P=0.001,ULBP1:P=0.022);用STAT3抑制剂(SH-4-54)处理K562/STAT3细胞后,MICA mRNA和蛋白表达水平升高(mRNA:P=0.001,蛋白:P=0.002),但ULBP1 mRNA和蛋白无明显改变(mRNA:P=0.137,蛋白:P=0.1905)。NK细胞对耐药K562/STAT3细胞的杀伤能力较K562/-细胞下降(P=0.002),但是STAT3抑制剂恢复了K562/STAT3细胞对NK细胞的杀伤敏感性(P=0.006)。结论:STAT3磷酸化介导了耐药白血病细胞抗凋亡,降低其对NK细胞的杀伤敏感性。STAT3抑制剂可以促进耐药白血病细胞凋亡并增加其对NK细胞的杀伤敏感性。Objective:To investigate the mechanisms of anti-apoptosis and immune evasion in drug-resistant leukemia cells mediated by STAT3,further to explore the possible mechanism of leukemia relapse caused by minimal residual.Methods:Drug-resistance leukemia cell line was established by transfecting pcDNA3.1-STAT3 into K562 cells(K562/STAT3).The expression of STAT3,BAX and NKG2D ligands(MICA and ULBP1)in K562/-cells,K562/STAT3 were detected by Western blot and/or RQ-PCR.Cells apoptosis and the killing effect of NK cells on leukemia cells were detected by flow cytometry.Results:The expression of the total STAT3,STAT3 phosphorylation in K562/STAT3 was significantly increased,and P-gp mRNA expression was increased also significantly(P<0.005).In K562/STAT3 cells,the expression of pro-apoptotic BAX(P=0.005)was significantly lower,and the number of apoptotic cells(P=0.002)induced by adriamycin was significantly decreased as compared with those in K562/-cells.After K562/STAT3 cells were treated by STAT3 inhibitor(SH-4-54),the expression of BAX mRNA(P=0.017)was significantly higher and the number of apoptotic cells(P=0.005)was significantly increased.The MICA and ULBP1 mRNA expression in K562/STAT3 cells was significantly lower than that in K562/-cells,and also for MICA and ULBP1 protein(MICA and ULPB1 mRNA:P<0.0001,MICA protein:P=0.001,ULPB1 protein:P=0.022).After K562/STAT3 cells were treated with STAT3 inhibitor(SH-4-54),the expression of MICA mRNA and protein was increased(mRNA:P=0.001,protein:P=0.002),but ULBP1 mRNA and protein showed no significantly change(mRNA:P=0.137,protein:P=0.1905).The cytotoxicity of NK cells to K562/STAT3 cells was susceptible as compared with K562/-(P=0.002),but the cytotoxicity of K562/STAT3 cells to NK cell could be recovered by STAT3 inhibitor(P=0.006).Conclusion:STAT3 phosphorylation can inhibits cell apoptosis and promotes cell immune escape.STAT3 inhibitors can promote the apoptosis of leukemia cells and increase their sensitivity to NK cells.

关 键 词：STAT3 白血病细胞 NK细胞 NKG2D配体 化疗耐药 

分 类 号：R733.71[医药卫生—肿瘤]