山西医科大学学报2023，Vol.54Issue(11)：1470-1476,7.DOI:10.13753/j.issn.1007-6611.2023.11.006
多发性骨髓瘤细胞分泌外泌体对破骨细胞分化的调控及其机制
Regulatory effect and its mechanism of multiple myeloma exosomes on osteoclast differentiation
摘要
Abstract
Objective To investigate the role and mechanism of multiple myeloma exosomes in regulating the expression of genes related to osteoclast differentiation.Methods To investigate the effect of multiple myeloma cells on osteoclast differentiation,the experiments were divided into control group(THP-1 cells),OPM2 group(THP-1+OPM2),U266 group(THP-1+U266),OPM2+GW4869 group(THP-1+OPM2+GW4869)and U266+GW4869 group(THP-1+U266+GW486).Exosomes secreted by OPM2 and U266 cells(OPM2 exo and U266 exo)were extracted,and then the exosomal structure was observed under transmission electron microscopy,and the exosomal marker proteins TSG101,Hsp70,and CD9 were identified by Western blot.To investigate the effect of exosomes on oste-oclast differentiation,the experiments were divided into OPM2 exo group(THP-1 cell+OPM2 exo),U266 exo group(THP-1 cell+U266 exo)and PBS group(THP-1 cell+PBS).RT-qPCR was used to detect the relative expression levels of NFATc1,oncogene c-Fos,and CTSK mRNA in all groups.Western blot was used to detect the relative expression levels of NFATc1,c-Fos,CTSK,LC3Ⅱ and P62 in all groups.Results The particle sizes of OPM2 exo and U266 exo were around 100 nm,with a"cupholder"like struc-ture,and OPM2 exo and U266 exo expressed marker proteins TSG101,Hsp70,and CD9.Compared with control group,the mRNA and protein expressions of NFATc1,c-Fos and CTSK,and the protein expression of LC3 Ⅱ were significantly upregulated in OPM2 group and U266 group(P＜0.001),while the expression of P62 protein was significantly downregulated(P＜0.001).The mRNA and protein expressions of NFATc1,c-Fos,CTSK in OPM2+GW4869 group were significantly lower than those in OPM2 group(P＜0.001),and the mRNA and protein expressions of NFATc1,c-Fos and CTSK in U266+GW4869 group were significantly lower than those in U266 group(P＜0.001).Compared with PBS group,the mRNA and protein expressions of NFATc1,c-Fos,CTSK were significantly upregulated in OPM2 exo group and U266 exo group(P＜0.001),LC3 Ⅱ protein expression was significantly upregulated(P＜0.001),and P62 protein expression was significantly downregulated(P＜0.001).Conclusion Multiple myeloma cell exosomes can induce osteoclast differentiation by promoting the cellular autophagy.关键词
多发性骨髓瘤/外泌体/破骨细胞分化/破骨细胞分化相关基因/自噬Key words
multiple myeloma/exosomes/osteoclast differentiation/osteoclast differentiation-related gene/autophagy分类
医药卫生引用本文复制引用
吴洋,吴镜,罗晓慧,张哲梅,侯金霞..多发性骨髓瘤细胞分泌外泌体对破骨细胞分化的调控及其机制[J].山西医科大学学报,2023,54(11):1470-1476,7.基金项目
兰州市科技计划项目(2020-ZD-25) （2020-ZD-25）
