1] Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China [2] Department of Hematology, Affiliated Hospital of Taishan Medical College, Taian 271000, China.
Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Acta Pharmacol Sin. 2014 Feb;35(2):230-8. doi: 10.1038/aps.2013.141. Epub 2013 Dec 30.
To investigate whether human multiple myeloma (MM) cells secrete microvesicles (MVs) and whether the MVs secreted from MM cells (MM-MVs) promote angiogenesis.
RPMI8226 human MM cells and EA.hy926 human umbilical vein cells were used. MVs isolated from RPMI8226 cells were characterized under laser confocal microscopy, electron microscopy and with flow cytometry. The fusion of MM-MVs and EA.hy926 cells was studied under confocal microscopy, and the transfer of CD138 to EA.hy926 cells was demonstrated with flow cytometry. The proliferation, invasion and tube formation of EA.hy926 cells in vitro were evaluated using MTT, transwell migration and tube formation assays, respectively. The vasculization of EA.hy926 cells in vivo was studied using Matrigel plug assay. The expression of IL-6 and VEGF was analyzed with PCR and ELISA.
MM-MVs from the RPMI 8226 cells had the characteristic cup-shape with diameter of 100-1000 nm. Most of the MM-MVs expressed phosphatidylserine and the myeloma cell marker CD138, confirming that they were derived from myeloma cells. After added to EA.hy926 cells, the MM-MVs transferred CD138 to the endothelial cells and significantly stimulated the endothelial cells to proliferate, invade, secrete IL-6 and VEGF, two key angiogenic factors of myeloma, and form tubes in vitro and in vivo.
Our results confirm the presence of MVs in MM cells and support the idea that MM-MVs are newfound mediators for myeloma angiogenesis and may serve as a therapeutic target to treat MM.
研究人多发性骨髓瘤（MM）细胞是否分泌微泡（MVs），以及 MM 细胞分泌的 MVs（MM-MVs）是否促进血管生成。
使用 RPMI8226 人 MM 细胞和 EA.hy926 人脐静脉细胞。通过激光共聚焦显微镜、电子显微镜和流式细胞术对从 RPMI8226 细胞中分离出的 MVs 进行了表征。通过共聚焦显微镜研究了 MM-MVs 与 EA.hy926 细胞的融合，并通过流式细胞术证实了 CD138 向 EA.hy926 细胞的转移。通过 MTT、Transwell 迁移和管形成测定分别评估了 EA.hy926 细胞在体外的增殖、侵袭和管形成。通过 Matrigel plugs 测定研究了 EA.hy926 细胞在体内的血管生成。通过 PCR 和 ELISA 分析了 IL-6 和 VEGF 的表达。
来自 RPMI8226 细胞的 MM-MVs 具有特征性的杯形，直径为 100-1000nm。大多数 MM-MVs 表达磷脂酰丝氨酸和骨髓瘤细胞标志物 CD138，证实它们来源于骨髓瘤细胞。加入 EA.hy926 细胞后，MM-MVs 将 CD138 转移到内皮细胞，并显著刺激内皮细胞增殖、侵袭、分泌 IL-6 和 VEGF，这两种都是骨髓瘤的关键血管生成因子，并在体外和体内形成管。
我们的结果证实了 MM 细胞中存在 MVs，并支持 MM-MVs 是骨髓瘤血管生成的新发现的介质的观点，并且可能作为治疗 MM 的治疗靶点。
