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Serum deprivation elevates the levels of microvesicles with different size distributions and selectively enriched proteins in human myeloma cells in vitro
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Serum deprivation elevates the levels of microvesicles with different size distributions and selectively enriched proteins in human myeloma cells in vitro
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Journal Article
Serum deprivation elevates the levels of microvesicles with different size distributions and selectively enriched proteins in human myeloma cells in vitro
2014
Overview
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.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ Biomedical and Life Sciences
/ Cell-Derived Microparticles - metabolism
/ Cell-Derived Microparticles - pathology
/ Culture Media, Serum-Free - metabolism
/ Electrophoresis, Gel, Two-Dimensional
/ Endothelial Cells - metabolism
/ Humans
/ Multiple Myeloma - metabolism
/ Multiple Myeloma - pathology
/ Myeloma Proteins - metabolism
/ Neovascularization, Physiologic
/ Original
/ Repressor Proteins - metabolism
/ Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
/ Vaccine
/ 体外选择
/ 尺寸分布
/ 微泡
/ 胎牛血清
/ 蛋白质谱
/ 骨髓瘤细胞
