Serum deprivation elevates the levels of microvesicles with different size distributions and selectively enriched proteins in human myeloma cells in vitro  

作　　者：Li SUN Hong-xiang WANG Xiao-jian ZHU Pin-hui WU Wei-qun CHEN Ping ZOU Qiu-bai LI Zhi-chao CHEN 

机构地区：[1]Institute of Hematology, Union Hospital, Tongii Medical College, Huazhong University of Science and Technology, Wuhan 430022, China [2]Department of Hematology, Wuhan Central Hospital, Wuhan 430022, China [3]First Clinical College, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China [4]Department of Central Laboratory, Wuhan Central Hospital, Wuhan 430022, China

出　　处：《Acta Pharmacologica Sinica》2014年第3期381-393,共13页中国药理学报（英文版）

摘　　要：Aim： To investigate the effects of serum deprivation （SD） on microvesicles （MVs） secreted from human myeloma cells and the implications for disease progression. Methods： RPMI 8226, U266, and KM3 human myeloma cells were incubated in medium containing 10% （non-SD） or 1% fetal bovine serum （SD） and MVs were isolated. The levels and size distribution of MVs were analyzed with flow cytometry. The protein profiles of MVs were studied using 2D SDS-PAGE, MALDI-TOF.MS, and Western blotting. NF-KB activation was analyzed using EMSA. Angiogenesis was examined in Eahy926 endothelial cells. Results： Exposure of RPMI 8226 ceils to SD for 24 h did not alter the number of apoptotic cells. However, SD increased the number of MVs from RPMI 8226, U266, and KM3 cells to 2.5-, 4.3-, and 3.8-fold, respectively. The size distribution of SD MVs was also significantly different from that of non-SD MVs. Three proteins ZNF224, SARM, and COBL in SD MVs were found to be up-regulated, which were involved in cell cycle regulation, signal transduotion and metabolism, respectively. Co-culture of SD MVs and RPMI 8226 cells increased NF-KB activation in the target RPMI 8226 cells. Furthermore, SD MVs from RPMI 8226 cells significantly increased the microtubule formation capacity of Eahy926 endottlelial cells compared with non-SD MVs. Conclusion： SD elevates the levels of microvesicles with different size distribution and selectively enriched proteins in human myeloma cells in vitro. The selectively enriched proteins, especially ZNF224, may play key roles in regulation of myeloma cells, allowing better adaptation to SD.Aim： To investigate the effects of serum deprivation （SD） on microvesicles （MVs） secreted from human myeloma cells and the implications for disease progression. Methods： RPMI 8226, U266, and KM3 human myeloma cells were incubated in medium containing 10% （non-SD） or 1% fetal bovine serum （SD） and MVs were isolated. The levels and size distribution of MVs were analyzed with flow cytometry. The protein profiles of MVs were studied using 2D SDS-PAGE, MALDI-TOF.MS, and Western blotting. NF-KB activation was analyzed using EMSA. Angiogenesis was examined in Eahy926 endothelial cells. Results： Exposure of RPMI 8226 ceils to SD for 24 h did not alter the number of apoptotic cells. However, SD increased the number of MVs from RPMI 8226, U266, and KM3 cells to 2.5-, 4.3-, and 3.8-fold, respectively. The size distribution of SD MVs was also significantly different from that of non-SD MVs. Three proteins ZNF224, SARM, and COBL in SD MVs were found to be up-regulated, which were involved in cell cycle regulation, signal transduotion and metabolism, respectively. Co-culture of SD MVs and RPMI 8226 cells increased NF-KB activation in the target RPMI 8226 cells. Furthermore, SD MVs from RPMI 8226 cells significantly increased the microtubule formation capacity of Eahy926 endottlelial cells compared with non-SD MVs. Conclusion： SD elevates the levels of microvesicles with different size distribution and selectively enriched proteins in human myeloma cells in vitro. The selectively enriched proteins, especially ZNF224, may play key roles in regulation of myeloma cells, allowing better adaptation to SD.

关 键 词：multiple myeloma MICROVESICLES nutrient deprivation cell adaptation NF-KB ZNF224 SARM COBL two-dimensional gelelectrophoresis 

分 类 号：Q516[生物学—生物化学] X592[环境科学与工程—环境工程]