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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 investigate the effect of 4-1BBL reverse signal on the growth of human multiple myeloma cell line U266 and its mechanism. Methods The expression of 4-1BBL on U266 cell line was detected by immunofluorescence staining and flow cytometry. Reverse transcription of 4-1BBL was induced by anti-human 4-lBBL monoclonal antibody (1F1). The proliferation of U266 cells was observed by cell counting and MTT assay IL-6 in U266 cells was detected by intracellular cytokine staining. The neutralizing antibody was used to block the biological effects of IL-6, and whether IL-6 was involved in 4-1BBL Reverse Signaling Promotes Proliferation of U266 Cells. Results High expression of 4-1BBL on the surface of U266 cells was observed. Cell counting and MTT assay showed that mAb 1F1 significantly promoted the growth of U266 cell line. Intracellular cytokine staining showed that 1F1 promoted U266 autocrine IL-6 secretion. Neutralization of IL-6 inhibited the proliferation of U266 cells induced by mAb 1F1. CONCLUSION: U266 cells express high levels of 4-1BBL molecules. Reverse signaling of 4-1BBL can promote the proliferation of U266 cells by increasing the production of IL-6. 4-1BB / 4-1BBL molecules may be important for immune intervention in multiple myeloma Targets.