小檗碱与维奈托克联用协同诱导获得性维奈托克耐药的人急性淋巴细胞白血病细胞凋亡

Combination of berberine and venetoclax synergistically induces apoptosis in human acute lymphoblastic leukemia cells with acquired venetoclax resistance

目的研究小檗碱(BBR)与维奈托克(VEN)联用诱导获得性VEN耐药的人急性淋巴细胞白血病RS4;11细胞(RS4;11-VENR)凋亡的作用及相关机制。方法通过将人急性淋巴细胞白血病RS4;11细胞长期暴露在VEN中获得RS4;11-VENR。基于ZIP模型评估BBR与VEN处理RS4;11-VENR的联用效果。通过流式细胞法检测两者联用对细胞凋亡率的影响。采用qRT-PCR法检测BBR与VEN处理后细胞抗凋亡蛋白基因表达水平的变化。Western blot法检测细胞凋亡相关蛋白表达水平以及Caspase-3的剪切情况。结果细胞凋亡诱导和增殖抑制实验证明VEN对RS4;11-VENR不敏感(IC_(50)>10μmol·L^(-1)),Western blot结果表明RS4;11-VENR对比野生型RS4;11细胞存在MCL-1上调;基于ZIP模型的联用评价表明12~50μmol·L^(-1)BBR与0.1~10μmol·L^(-1)VEN协同杀伤RS4;11-VENR,流式结果显示25μmol·L^(-1)BBR与1μmol·L^(-1)VEN联用协同诱导细胞凋亡;qRT-PCR实验结果显示25~30μmol·L^(-1)BBR单独处理细胞致MCL-1、BCL-XL基因表达水平降低,Western blot结果显示BBR单独处理降低细胞MCL-1、BCL-XL蛋白水平。结论BBR与VEN联用协同杀伤RS4;11-VENR,BBR可能通过降低RS4;11-VENR的MCL-1、BCL-XL蛋白水平,增加细胞对VEN敏感性从而诱导凋亡发生。

Objective To determine the effect and mechanism of berberine(BBR)combined with venetoclax(VEN)in inducing apoptosis in human acute lymphoblastic leukemia RS4 and 11 cells with acquired drug resistance to VEN(RS4;11-VENR).Methods RS4;11-VENR were obtained by long-term exposure to human acute lymphoblastic leukemia RS4;11 cells to VEN.Evaluation of the effect of combination of BBR and VEN on RS4;11-VENR based on ZIP model was evaluated.The effect of the combination on the apoptosis rate was detected by flow cytometry.qRT-PCR method was used to detect the changes of anti-apoptotic protein gene expression in cells treated with BBR and VEN combination.The expression of apoptosis-related proteins and the splicing of Caspase-3 were detected by Western blot.Results Apoptosis induction and proliferation inhibition showed that VEN Scientifwas not sensitive to RS4;11-VENR(IC_(50)>10μmol·L^(-1))and Western blot showed that MCL-1 was up-regulated by RS4;11-VENR as compared with wild-type RS4;11 cells.The combination evaluation based on ZIP model showed that 12~50μmol·L^(-1) BBR and 0.1~10μmol·L^(-1) VEN synergistically killed RS4;11-VENR;the flow cytometry showed that 25μmol·L^(-1) BBR combined with 1μmol·L^(-1) VEN synergistically induced the apoptosis;the qRT-PCR assay showed that 25~30μmol·L^(-1) BBR alone decreased the expression of MCL-1 and BCL-XL genes;and Western blot showed that BBR alone decreased the protein levels of MCL-1 and BCLXL.Conclusion BBR combined with VEN synergistically kills RS4;11-VENR.BBR may induce the apoptosis by reducing the levels of MCL-1 and BCL-XL proteins and increasing the sensitivity of RS4;11-VENR to VEN.