慢性淋巴细胞白血病中BIM诱导伊布替尼耐药机制研究  

Mechanism of BIM-induced ibrutinib resistance in chronic lymphocytic leukemia

作　　者：张加乐 潘必慧 吴佳竹[1] 孔祎琳 王莉[1] 徐卫[1] Zhang Jiale;Pan Bihui;Wu Jiazhu;Kong Yilin;Wang Li;Xu Wei(Department of Hematology,The First Affiliated Hospital of Nanjing Medical University(Jiangsu Province Hospital),Nanjing 21009,China)

机构地区：[1]南京医科大学第一附属医院(江苏省人民医院)血液科,南京210029

出　　处：《中华血液学杂志》2025年第2期152-160,共9页Chinese Journal of Hematology

基　　金：国家自然科学基金(82370194);国家自然科学基金青年基金(82200210);南京医科大学附属第一医院青年基金培育计划(PY2021026)。

摘　　要：目的探究BCL2家族蛋白BIM在慢性淋巴细胞白血病(CLL)伊布替尼耐药中的作用,分析其对凋亡和自噬的调控机制。方法通过RNA测序检测CLL患者样本、CLL细胞株(MEC1细胞株)和耐药细胞株(MR)中BCL2家族表达变化。蛋白印迹法分析药物敏感细胞株凋亡时BIM蛋白表达变化,并用shRNA敲低BIM探讨其对增殖和凋亡的影响。利用RNA测序和自噬抑制剂氯喹观察MR自噬水平变化。结果CLL原代细胞及MECI细胞株耐药前后,BIM表达显著下调(P<0.0001)。在CLL细胞中敲低BIM可抑制由伊布替尼诱导的细胞凋亡(均P<0.05)。此外,在MR中可以观察到保护性自噬水平的提高,通过氯喹及小干扰RNA抑制自噬反应后可促进细胞凋亡。BIM敲低细胞株中LC3-Ⅱ蛋白表达增加(P<0.01),提示BIM降低可介导自噬激活。结论BIM低表达是CLL中伊布替尼耐药的重要因素,其通过激活保护性自噬促进耐药,这为改善CLL治疗提供了新靶点。ObjectiveTo investigate the relationship between the BCL2 family protein BIM and ibrutinib resistance in chronic lymphocytic leukemia(CLL)and to analyze its regulatory mechanisms on apoptosis and autophagy.MethodsRNA sequencing(RNA-seq)was used to examine changes in the expression of BCL2 family proteins in samples from patients with CLL,MEC1 cell lines,and ibrutinib-resistant cell lines(MR).Western blot was used to analyze changes in BIM protein expression during apoptosis in MR.shRNA knockdown was used to assess the effects of BIM on cell proliferation and apoptosis.RNA-seq and the autophagy inhibitor chloroquine treatment were used to study autophagy-related changes in MR.ResultsBIM expression was significantly downregulated before and after drug resistance in CLL primary cells and MEC1 cell lines(P<0.0001).Knockdown of BIM in CLL cells inhibited ibrutinib-induced apoptosis and promoted cell proliferation(P<0.05 for both).In addition,protective autophagy was increased in MR and apoptosis was increased after administration of chloroquine and small interfering RNA.The increased expression of LC3-Ⅱprotein in BIM-knockdown cell lines(P<0.01)suggested that reduction of BIM may mediate autophagy activation.ConclusionDownregulation of BIM may be a key factor in promoting ibrutinib resistance in CLL by activating protective autophagy.These findings provided a potential target for improving CLL treatment.

关 键 词：白血病 淋巴细胞 慢性 B细胞 伊布替尼 耐药 BIM 自噬 

分 类 号：R733.72[医药卫生—肿瘤]