Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China.
Mol Med Rep. 2013 Jul;8(1):272-6. doi: 10.3892/mmr.2013.1458. Epub 2013 May 8.
The aims of the current study were to determine whether 786‑0 renal cancer cell‑derived exosomes promote human umbilical vein endothelial cells (HUVECs) to form tubular structures and to uncover the underlying mechanisms associated with this process. Exosomes were extracted and purified using ultrafiltration and sucrose gradient centrifugation and characterized by transmission electron microscopy. Tubular structure formation was observed using the matrigel tubular assay. In addition, an adenovirus vector was used to transfect the hepatocyte cell adhesion molecule (hepaCAM) gene into renal cancer 786‑0 cells. The expression of hepaCAM and vascular endothelial growth factor (VEGF) mRNA and protein was determined by reverse transcription‑polymerase chain reaction and western blot analysis, respectively. Tumor cell‑derived exosomes were observed to significantly increase tubular formation in HUVECs. Following transfection with the hepaCAM gene, VEGF expression in 786‑0 cells was markedly decreased. In HUVECs, exosome treatment increased VEGF mRNA and protein expression, while hepaCAM expression was only decreased at the protein level. In the present study, renal cancer 786‑0 cell‑derived exosomes significantly promoted angiogenesis via upregulation of VEGF expression in HUVECs, which may be induced by the downregulation of hepaCAM.
本研究旨在确定 786-0 肾癌细胞衍生的外泌体是否能促进人脐静脉内皮细胞(HUVEC)形成管状结构,并揭示与该过程相关的潜在机制。使用超滤和蔗糖密度梯度离心法提取和纯化外泌体,并通过透射电子显微镜进行表征。使用基质胶管状测定法观察管状结构的形成。此外,还使用腺病毒载体将肝细胞黏附分子(hepaCAM)基因转染至肾癌细胞 786-0 中。通过逆转录-聚合酶链反应和 Western blot 分析分别测定 hepaCAM 和血管内皮生长因子(VEGF)mRNA 和蛋白的表达。观察到肿瘤细胞衍生的外泌体可显著增加 HUVEC 中的管状形成。转染 hepaCAM 基因后,786-0 细胞中 VEGF 的表达明显降低。在 HUVEC 中,外泌体处理可增加 VEGF mRNA 和蛋白的表达,而 hepaCAM 的表达仅在蛋白水平下降。在本研究中,肾癌细胞 786-0 衍生的外泌体通过上调 HUVEC 中 VEGF 的表达显著促进血管生成,这可能是通过下调 hepaCAM 诱导的。