IL-6通过STAT3/Notch信号通路调控多发性骨髓瘤耐药细胞株对硼替佐米的化疗敏感性  

IL-6 Regulates the Chemosensitivity of Drug-Resistant Multiple Myeloma Cell Lines to Bortezomib through STAT3/Notch Signaling Pathway

作　　者：刘莹[1] 隋靖喆[2] 朱丽华[3] 戴益 蕫海群 程鹏[1] LIU Ying;SUI Jing-Zhe;ZHU Li-Hua;DAI Yi;DONG Hai-Qun;CHENG Peng(Department of Hematology,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Department of Clinical Laboratory,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Department of Health Management,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China;The First Clinical Medical College of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学第一附属医院血液内科,广西南宁530021 [2]广西医科大学第一附属医院临床检验科,广西南宁530021 [3]广西医科大学第一附属医院健康管理部,广西南宁530021 [4]广西医科大学第一临床医学院,广西南宁530021

出　　处：《中国实验血液学杂志》2022年第5期1474-1481,共8页Journal of Experimental Hematology

基　　金：广西自然科学基金青年科学基金项目(2020GXNSFBA297061);广西壮族自治区卫生健康委员会自筹经费科研计划课题(Z20201124)。

摘　　要：目的:探讨白细胞介素-6(IL-6)调控多发性骨髓瘤(MM)耐药细胞株对硼替佐米(bortezomib,BTZ)的化疗敏感性及相关作用机制。方法:收集临床BTZ耐药MM患者治疗前后的外周血样本,体外培养人KM3和KM3/BTZ细胞,ELISA法检测MM患者外周血、KM3和KM3/BTZ细胞中IL-6的含量,CCK-8法检测KM3和KM3/BTZ细胞对BTZ的药物敏感性。将KM3/BTZ细胞分为KM3/BTZ对照组(细胞正常培养48 h),IL-6中和抗体Anti-IL-6组(500 ng/ml Anti-IL-6处理细胞48 h),BTZ组(300 ng/ml BTZ处理细胞48 h),BTZ+Anti-IL-6组(300 ng/ml BTZ和500 ng/ml Anti-IL-6处理细胞48 h)。采用CCK-8法检测KM3/BTZ细胞增殖活性,流式细胞术检测KM3/BTZ细胞周期分布情况,Annexin V-FITC/PI双染法检测KM3/BTZ细胞凋亡,实时荧光定量PCR(q RT-PCR)检测KM3/BTZ细胞IL-6、Notch1、信号转导和转录激活因子3(STAT3)mRNA表达水平,蛋白质印迹法(Western blot)检测KM3/BTZ细胞IL-6、Notch1、STAT3蛋白表达水平。结果:治疗后BTZ耐药MM患者外周血中IL-6含量显著高于治疗前(P<0.05)。KM3/BTZ细胞中IL-6含量显著高于KM3细胞(P<0.05)。KM3/BTZ细胞对BTZ的敏感性显著低于KM3细胞(P<0.05),耐药指数(RI)为19.62。Anti-IL-6和BTZ均可抑制KM3/BTZ细胞增殖、阻滞细胞周期、诱导细胞凋亡(P<0.05);与单一药物处理相比,Anti-IL-6和BTZ联合处理对KM3/BTZ细胞的作用更为明显(P<0.05),且显著下调KM3/BTZ细胞IL-6、Notch1、STAT3 mRNA和蛋白表达(P<0.05)。结论:拮抗IL-6可以增加BTZ的化疗敏感性,IL-6可能通过STAT3/Notch信号通路降低MM细胞对BTZ的敏感性。Objective: To investigate the effect of interleukin-6(IL-6) on the chemosensitivity of drug-resistant multiple myeloma(MM) cell lines to bortezomib(BTZ) and its mechanism. Methods: Peripheral blood samples were collected from patients with BTZ-resistant MM before and after treatment. Human MM cell lines KM3 and KM3/BTZ were cultured in vitro. ELISA was used to detect the content of IL-6 in peripheral blood of MM patients, KM3 and KM3/BTZ cells. CCK-8 assay was used to detect the drug sensitivity of KM3 and KM3/BTZ cells to BTZ. KM3/BTZ cells were divided into KM3/BTZ control group(normal culture for 48 h), IL-6 neutralizing antibody Anti-IL-6 group(500 ng/ml Anti-IL-6 treated for 48 h), BTZ group(300 ng/ml BTZ treated for 48 h), BTZ + Anti-IL-6 group(300 ng/ml BTZ and 500 ng/ml Anti-IL-6 treated for 48 h). The proliferation activity of KM3/BTZ cells was detected by CCK-8 assay. The cell cycle distribution of KM3/BTZ cells was detected by flow cytometry. The apoptosis of KM3/BTZ cells was detected by Annexin V-FITC/PI double staining. The mRNA expression levels of IL-6, Notch1, signal transducer and activator of transcription 3(STAT3) in KM3/BTZ cells were detected by real-time fluorescent quantitative PCR(qRT-PCR), and the protein expression levels of IL-6, Notch1, STAT3 in KM3/BTZ cells were detected by Western blot. Results: The level of IL-6 in peripheral blood of patients with BTZ-resistant MM after treatment was significantly higher than that before treatment(P<0.05). The level of IL-6 in KM3/BTZ cells was significantly higher than that in KM3 cells(P<0.05). The sensitivity of KM3/BTZ cells to BTZ was significantly lower than that of KM3 cells(P<0.05), and the resistance index(RI)was 19.62. Anti-IL-6 and BTZ could inhibit the proliferation of KM3/BTZ cells, block cell cycle, and induce apoptosis(P<0.05). Compared with single drug treatment, the combined effect of Anti-IL-6 and BTZ was more obvious on KM3/BTZ cells(P<0.05), and significantly down regulated the mRNA and protein expression of IL-6, Notch1 and S

关 键 词：白细胞介素-6 多发性骨髓瘤 硼替佐米 化疗敏感性 

分 类 号：R733.3[医药卫生—肿瘤]