摘要
目的:研究FLT3-ITD突变体ITD插入长度对细胞增殖能力、凋亡及靶向抑制剂敏感性的影响,为临床分级治疗FLT3-ITD突变型急性髓系白血病提供数据参考。方法:从先前解析出的ITD序列中挑选3个位置相同或相邻的不同大小的ITD序列,构建FLT3-ITD真核表达质粒;利用慢病毒包被系统建立FLT3-ITD稳定表达32D细胞株;检测FLT3-ITD细胞株细胞增殖情况,经靶向抑制剂AC220处理后的其增殖抑制及凋亡情况。结果:成功构建了ITD1、ITD2和ITD3三株FLT3-ITD稳定表达32D细胞株;ITD1、ITD2和ITD3细胞株在无IL3因子培养基生长条件下48 h后的增殖倍数分别是2.3、3.7和4.3倍;在FLT3小分子抑制剂AC220加药条件下,ITD1、ITD2和ITD3细胞株增殖抑制IC50值分别是0.183、0.446和0.836 nmol/L,凋亡率分别是88.6%、34.2%和16.1%。结论:插入ITD长度长的FLT3-ITD突变体表达细胞在增殖能力和耐受靶向抑制剂AC220的能力方面均增强。
Objective: To study the effects of FLT3-ITD length on 32 D cell proliferation,apoptosis and sensitivity to FLT3 inhibitor,so as to provide references for stepwise therapy of FLT3-ITD mutated acute myeloid leukemia patients.Methods: Three different FLT3-ITD mutants with same or adjacent insert sites were selected and constructed in an eukaryotic expression vector. FLT3-ITD mutants stably expressed 32 D cell strains were selected with the help of lentivirus system and IL3 free cell culture medium. The proliferation and apoptosis of 32 D cell strains after AC220 treatment were detected. Results: FLT3-ITD mutants( ITD1,ITD2 and ITD3) stably expressed 32 D cell strains were constructed successfully. In the absence of IL3 factor,the proliferation number of ITD1,ITD2 and ITD3 cell strains were mounted up to 2. 3 folds,3. 7 folds,and 4. 3 folds after 48 hours,respectively. Under the exposure of FLT3 inhibitor AC220,the IC50 values was 0. 183,0. 446 and 0. 836 nmol/L,and apoptosis rates was 88. 6%,34. 2% and16. 1%,respectively. Conclusion: FLT3-ITD mutant expressed cell strains with longer ITD show higher capacity of proliferation and higher tolerance to AC220 treatment.
作者
刘松柏
董豪杰
王隽
邱桥成
薛胜利
李凌
LIU Song-Bai;DONG Hao-Jie;WANG Jun;QIU Qiao-Cheng;XUE Sheng-Li;LI Ling(Suzhou Key Laboratory for Medical Biotechnology,Suzhou Vocational Health College,Suzhou 215009,Jiangsu Province,China;Department of Hematological Malignancies Translational Science,Beckman Research Institute,City of Hope National Medical Center,Duarte,CA 91010,USA;Department of Hematology,The First Affiliated Hospital of Soochow University,Jiangsu Institute of Hematology,Suzhou 215006,Jiangsu Province,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2021年第4期1034-1038,共5页
Journal of Experimental Hematology
基金
国家自然科学基金项目(31701198,81970138)
江苏省自然科学基金(BK20170386)
江苏省高层次卫生人才“六个一工程”拔尖人才科研项目(LGY2018087)
江苏高校“青蓝工程”优秀青年骨干教师培养项目(2017)
苏州卫生职业技术学院科技创新团队(SZWZYTD201804)。
