芦可替尼和地西他滨协同抑制TET2敲降的HEL细胞生长作用  

Study on synergistic inhibitory effect of ruxolitinib and decitabine on growth of HEL cells with TET2 knockdown

作　　者：付莉霞 王颖韶 肖方楠 宋濬哲 白姣姣 周圆 白洁 FU Li-xia;WANG Ying-shao;XIAO Fang-nan;SONG Jun-zhe;BAI Jiao-jiao;ZHOU Yuan;BAI Jie(The Second Hospital of Tianjin Medical University, Tianjin 300211, China;State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China)

机构地区：[1]天津医科大学第二医院,天津300211 [2]中国医学科院血液病医院(中国医学科学院血液学研究所),实验血液学国家重点实验室,国家血液系统疾病临床医学研究中心,天津300020

出　　处：《中国药理学通报》2021年第7期985-990,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81770128,81970120)。

摘　　要：目的探索地西他滨和芦可替尼对TET2敲降HEL细胞的杀伤及协同作用。方法利用shRNA构建TET2敲降的HEL细胞系;利用CCK-8法检测TET2敲降HEL细胞的增殖情况;通过测定半数致死量(IC_(50))和克隆形成实验评价TET2敲降HEL对芦可替尼和地西他滨的敏感性,并结合Chou-Talalay法计算地西他滨和芦可替尼在TET2敲降HEL细胞中的协同抑制效应。结果成功建立TET2敲降HEL细胞系两株,TET2敲降后HEL细胞增殖能力增加。72 h的药物敏感试验证实TET2敲降HEL细胞对地西他滨耐药。另外,克隆形成实验表明,TET2敲降后HEL细胞对芦可替尼和地西他滨的敏感性均有所下降。芦可替尼和地西他滨在TET2敲降HEL细胞中有显著的协同抑制效应。结论地西他滨与芦可替尼协同作用提示其可能是加速急变期JAK2^(V617F)和TET2双突变MPN患者有效的联合用药方案。Aim To explore the cytotoxic and synergistic effects of decitabine and ruxolitinib on HEL cells with TET2 knockdown.Methods Stable TET2 knockdown by shRNA was established in HEL cell line.The change of cell proliferation was measured by CCK-8 assay.The median lethal dose(IC_(50))and colony formation assay were used to evaluate the cytotoxic effects of decitabine and ruxolitinib,the synergistic effects of which was further analyzed by Chou-Talalay method.Results The inhibition of TET2 increased the proliferative capacity of HEL cells.HEL cell lines became resistant to decitabine following shRNA-mediated TET2 inactivation.Colony formation assay showed that the drug sensitivity of decitabine and ruxolitinib both decreased in TET2 knockdown HEL cells.The synergistic inhibitory effects of ruxolitinib and decitabine on TET2 knockdown HEL cells were observed.Conclusion The combination of ruxolitinib and decitabine may be an effective therapeutic strategy for accelerated or blast phase MPN patients with JAK2^(V617F) and TET2 mutations.

关 键 词：骨髓增殖性肿瘤 TET2 JAK2^(V617F) 芦可替尼 地西他滨 联合用药 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R394.2[医药卫生—基础医学]