AntiCancer, Inc., 7917 Ostrow Street, San Diego, CA 92111, USA.
Anticancer Res. 2013 Aug;33(8):3021-5.
αv Integrin is involved in various steps of cancer metastasis. In this report, we describe real-time imaging of αv integrin molecular dynamics in human 143B osteosarcoma cells in vitro and in vivo. We first generated osteosarcoma cells expressing αv integrin green fluorescent protein (GFP) by transfection of an αv integrin GFP fusion vector (pCMV6-AC-ITGAV-GFP) into 143B cells. Confocal laser-scanning microscopy demonstrated that αv integrin immunofluorescence staining co-localized with αv integrin-GFP fluorescence in 143B cells. When αv integrin-GFP-expressing 143B osteosarcoma cells were seeded on a dish coated with fibronectin, which is bound by αv integrin, punctate αv integrin-GFP was observed by confocal laser-scanning microscopy. When the 143B αv integrin-GFP cells were seeded onto uncoated plastic, αv integrin-GFP was diffuse within the cells. When αv integrin-GFP 143B osteosarcoma cells (1×10(6)) were orthotopically transplanted into the tibia of nude mice, the cells aligned along the collagen fibers within the tumor and had punctuate expression of αv integrin-GFP. In the orthotopic model, the invading osteosarcoma cells had punctate αv integrin-GFP in the muscle tissue at the primary tumor margin. These results show that αv integrin-GFP enables the imaging of the molecular dynamics of αv integrin in osteosarcoma cells in vitro and in vivo.
αv 整合素参与癌症转移的多个步骤。在本报告中，我们描述了体外和体内人 143B 骨肉瘤细胞中 αv 整合素分子动力学的实时成像。我们首先通过将 αv 整合素 GFP 融合载体 (pCMV6-AC-ITGAV-GFP) 转染到 143B 细胞中，生成表达 αv 整合素 GFP 的骨肉瘤细胞。共聚焦激光扫描显微镜显示，143B 细胞中 αv 整合素免疫荧光染色与 αv 整合素-GFP 荧光共定位。当表达 αv 整合素-GFP 的 143B 骨肉瘤细胞接种在涂有纤维连接蛋白的培养皿上时，通过共聚焦激光扫描显微镜观察到点状的 αv 整合素-GFP。当 143B αv 整合素-GFP 细胞接种在未涂覆的塑料上时，αv 整合素-GFP 在细胞内弥散。当 143B αv 整合素-GFP 骨肉瘤细胞 (1×10(6)) 原位移植到裸鼠胫骨中时，细胞沿肿瘤内的胶原纤维排列，并表现出点状的 αv 整合素-GFP 表达。在原位模型中，侵袭性骨肉瘤细胞在原发性肿瘤边缘的肌肉组织中具有点状的 αv 整合素-GFP。这些结果表明，αv 整合素-GFP 能够在体外和体内成像骨肉瘤细胞中 αv 整合素的分子动力学。
