4-1BBL逆向信号对人多发性骨髓瘤细胞株U266的促生长作用及其机制研究  

Study on 4-1BBL Reverse Signaling Enhance Proliferation in Human Multiple Myeloma Cell Line U266 and Its Mechanism

作　　者：汪娟[1] 居颂光[2] 傅 

机构地区：[1]苏州大学附属第一医院血液科江苏省血液研究所卫生部血栓与止血重点实验室,江苏苏州215006 [2]苏州大学医学部基础医学与生物科学学院免疫学系,江苏苏州215123

出　　处：《苏州大学学报（医学版）》2011年第5期731-734,共4页Suzhou University Journal of Medical Science

基　　金：江苏省医学重点人才项目资助(RC2007074);苏州市科技计划项目资助(YJS0914);国家自然基金青年科学基金项目资助(81000912);青蓝工程资助(2010)

摘　　要：目的探讨4-1BBL逆向信号对人多发性骨髓瘤细胞株U266的促生长作用及其机制。方法采用免疫荧光标记和流式细胞术检测U266细胞株表面4-1BBL分子的表达;采用抗人4-lBBL单抗(1F1)激发4-1BBL逆向信号,通过细胞计数及MTT法观察U266细胞增殖;通过细胞内细胞因子染色检测U266细胞内白细胞介素-6(IL-6)的合成,并利用中和性抗体阻断IL-6的生物学作用,观察IL-6是否参与了4-1BBL逆向信号对U266细胞的促增殖作用。结果 U266细胞表面高表达4-1BBL分子;细胞计数及MTT实验均表明单抗1F1明显促进U266细胞系生长;细胞内细胞因子染色示1F1促进U266自分泌IL-6增多。中和IL-6能够抑制单抗1F1对U266细胞促增殖作用。结论 U266细胞株高表达4-1BBL分子;4-1BBL逆向信号通过增加IL-6的产生,促进U266细胞的增殖;4-1BB/4-1BBL分子可能是对多发性骨髓瘤进行免疫干预的重要靶点。Objective To explore the biological effect of 4-1BBL reverse signaling on human multiple myeloma cell line U266 and its underlying mechanism.Methods The expression of 4-1BBL on U266 cell was detected by immunofluorescence staining and flowcytometric assay.Triggered by anti-4-1BBL mAb（1F1）,the proliferation of U266 cell was measured by cell counting and MTT assay.Intracellular cytokine staining and flowcytometry were performed to evaluate the interleukin-6（IL-6） production of U266 cells.To observe whether IL-6 participated in the proliferation of U266 cells triggered by 4-1BBL reverse signaling,neutralizing anti-IL-6 mAb was added to the culture medium to block the biology activity of IL-6.Results 4-1BBL molecules were highly expressed on U266 cells.Both cell counting and MTT assay showed that mAb 1F1 promoted U266 cell proliferation remarkably.Intracellular cytokine staining demonstrated mAb1F1 increased in the IL-6 production of U266 cells.Neutralizing anti-IL-6 mAb inhibited U266 cell proliferation stimulated by mAb 1F1.Conclusion 4-1BBL is constitutively expressed on U266 cell,4-1BBL reverse signaling can promote the proliferation of U266 cell through increasing the production of IL-6,and 4-1BB/4-1BBL molecules may be valuable targets for the immunotherapy of multiple myeloma.

关 键 词：4-1BBL 逆向信号 多发性骨髓瘤 白细胞介素-6 

分 类 号：R551.2[医药卫生—血液循环系统疾病] R733.3[医药卫生—内科学]