重组人BAFF对多发性骨髓瘤细胞BAFF-R表达调控的初步研究

Preliminary study on the regulation of recombinant human BAFF in the BAFF-R expression in multiple myeloma

目的 :研究多发性骨髓瘤(multiple myeloma,MM)中重组人B淋巴细胞刺激因子(recombinant human B lymphocyte stimulator,rh BAFF)对其特异性受体(B lymphocyte stimulator receptor,BAFF-R)基因、蛋白水平表达的影响;并进一步研究重组人BAFF与JNK信号通路之间的作用与BAFF-R之间的关系。方法:应用RT-PCR、酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)、细胞免疫荧光技术检测BAFF-R在MM细胞KM3中的表达、功能及调节;Western Blot分析BAFF/BAFF-R对NF-κB信号通路的激活情况。结果:细胞免疫荧光结果显示BAFF-R定位在KM3细胞的细胞膜上;重组人BAFF(50 ng/m L)能够促进BAFF-R m RNA和蛋白的表达;可溶性BAFF(50 ng/m L)促进p-JNK表达的上调,且随作用时间的延长而增加。结论 :重组人BAFF对BAFF-R表达有重要的影响,JNK信号通路可能参与了MM的发生发展过程。

Objective ： To investigate the regulation of recombinant human B lymphocyte stimulator（ rh BAFF） in the expression of BAFF-R m RNA and protein in multiple myeloma cells, and study the relationship between soluble BAFF and JNK signaling pathway. Methods： BAFF-R m RNA and protein expression, function and regulation were detected by real-time PCR, enzyme linked immunosorbent assay（ELISA） analysis, and immunofluorescence in KM3 cells. Western Blot was used to verify the selective capacity of BAFF/BAFF-R in activating the NF-κB pathway. Results： The immunofluorescence results showed that BAFF-R positioned on KM3 cell membrane. Rh BAFF increased BAFF-R m RNA and protein expression in KM3 cells. It was also found that rh BAFF induced the expression of p-JNK, and p-JNK increases as the exposure time increases.Conclusions： Rh BAFF has a significant impact on the expression of BAFF-R. JNK signaling pathway may be involved in the development of multiple myeloma.