Yang Hua, Fang Guofu, Huang Xinping, Yu Jie, Hsieh Chia-Ling, Grossniklaus Hans E
Department of Ophthalmology, Emory University School of Medicine, Atlanta, GA, USA.
Melanoma Res. 2008 Apr;18(2):95-103. doi: 10.1097/CMR.0b013e3282f628df.
The purpose of the study is to develop a mouse ocular melanoma model with human uveal melanoma cells that forms hepatic micrometastases. Human uveal melanoma Mel290 cells were transfected with a lentiviral-enhanced green fluorescent protein (EGFP) expression vector. Proliferation assays were performed by comparing Mel290-EGFP and Mel290 cells. After stable expression of EGFP and proliferation was ascertained, 1 x 10 Mel290-EGFP cells were introduced into NU/NU mice by posterior compartment (PC) inoculation or tail vein injection. Control groups were inoculated or injected with Mel290 cells. Ocular and hepatic frozen sections were examined by fluorescence microscopy, and the number of hepatic micrometastases was determined. EGFP expression was observed at 24 h after transfection. At 72 h after transfection, more than 70% of Mel290 cells expressed EGFP. At 45 days (six passages), 90% of Mel290 cells stably expressed EGFP. Histologic examination showed that Mel290-EGFP cells formed hepatic micrometastases after either PC inoculation or tail vein injection. A significant difference in the number of hepatic micrometastases between PC inoculation and tail vein injection (P<0.01) was observed. Mel290-EGFP cells stably expressed green fluorescent protein in vitro at 45 days (six passages). These cells formed hepatic micrometastases in NU/NU mice after PC inoculation or tail vein injection, with significantly more micrometastases developing in the PC inoculation model than after tail vein injection.
本研究的目的是利用人葡萄膜黑色素瘤细胞建立一种能形成肝微转移的小鼠眼黑色素瘤模型。用人葡萄膜黑色素瘤Mel290细胞转染慢病毒增强型绿色荧光蛋白(EGFP)表达载体。通过比较Mel290 - EGFP细胞和Mel290细胞进行增殖测定。在确定EGFP稳定表达和细胞增殖后,将1×10个Mel290 - EGFP细胞通过后房(PC)接种或尾静脉注射引入NU/NU小鼠体内。对照组接种或注射Mel290细胞。通过荧光显微镜检查眼和肝的冰冻切片,并确定肝微转移灶的数量。转染后24小时观察到EGFP表达。转染后72小时,超过70%的Mel290细胞表达EGFP。在45天(传代6次)时,90%的Mel290细胞稳定表达EGFP。组织学检查显示,Mel290 - EGFP细胞在PC接种或尾静脉注射后均形成肝微转移灶。观察到PC接种和尾静脉注射后肝微转移灶数量有显著差异(P<0.01)。Mel290 - EGFP细胞在45天(传代6次)时在体外稳定表达绿色荧光蛋白。这些细胞在PC接种或尾静脉注射后在NU/NU小鼠体内形成肝微转移灶,PC接种模型中形成的微转移灶明显多于尾静脉注射后形成的微转移灶。