目的：通过玻璃体注射经荧光标记的Y79细胞建立视网膜母细胞瘤BALB/c-nu小鼠模型，并以视网膜母细胞瘤常用化疗药物Melphalan治疗组作为阳性对照，通过荧光成像技术验证，以期为视网膜母细胞瘤治疗靶点或药物的非临床药效评价提供参考。

方法：BALB/c-nu小鼠玻璃体内注射GFP转染Y79细胞(1.0×10⁷ cell/mL，3 μL)造模，造模27 d根据活体成像荧光值随机分为模型对照组和不同剂量Melphalan组(1、3、10 μg/eye组)，玻璃体单次给药，每天观察眼部症状； 于造模后12、20、29、35、42、48、55、76、83 d对造模眼/给药眼进行裂隙灯检查，造模12、20、27、41、48、55、62、69、76、83 d行活体成像检查； 末次处理摘取眼球、大脑和小脑组织进行组织病理学检查。

结果：造模6 d开始，动物眼内可见云状物质，模型对照组小鼠眼云状物后期占据整个眼球，并伴有血管不规则生长； 造模27 d后动物眼均检测到荧光值，随造模时间延长，荧光值持续增加，造模69-83 d，肿瘤荧光值达峰值。组织学检查见模型对照组眼内瘤细胞重度增殖，1例模型动物大脑可见肿瘤细胞。Melphalan 10 μg/eye组在给药后17 d荧光值显著降低； Melphalan 3 μg/eye组给药后59 d荧光值显著抑制，Melphalan组动物脑组织均未见肿瘤细胞。

结论：BALB/c-nu小鼠玻璃体注射Y79/pCDH-LUC-copGFP细胞后，可观测到明显眼部病变及肿瘤细胞在眼部的增殖。同时，Melphalan干预后肿瘤细胞显著抑制，且呈剂量依赖性，表明视网膜母细胞瘤小鼠模型构建成功。

METHODS: BALB/c-nu mice were intravitreous injected with GFP transfected Y79 cells(1.0×10⁷ cell/mL, 3 μL)to establish the model. On the 27th day, the mice were randomly divided into model control group and different doses of Melphalan groups(1, 3, 10 μg/eye groups)according to the fluorescence value of in vivo imaging, with vitreous body single administrated and ocular symptoms observed daily. Slit-lamp examination was performed at 12, 20, 29, 35, 42, 48, 55, 76, and 83 d after modeling. In vivo imaging was performed on 12, 20, 27, 41, 48, 55, 62, 69, 76, and 83 d. At the last treatment, the eyeball, brain and cerebellum tissues were removed for histopathological examination.

RESULTS: From the sixth day of modeling, cloud-like substances could be seen in the eyes of the animals, and the cloud-like substances occupied the whole eyeball of the mice in the model control group at the later stage, accompanied by irregular growth of blood vessels. After 27 days of modeling, the fluorescence value was detected in all the animals, and the fluorescence value continued to increase with the extension of modeling time. The fluorescence value of the tumor reached the peak after 69-83 days of modeling. Histological examination showed severe proliferation of intraocular tumor cells in the model control group, and tumor cells were observed in the brain of 1 model animal. In the 10 μg/eye Melphalan group, the fluorescence value was significantly decreased at 17 d after administration. The fluorescence value of the 3 μg/eye Melphalan group was significantly inhibited at 59 d after administration. No tumor cells were found in the brain tissue of animals in all Melphalan groups.

CONCLUSION: After vitreous injection of Y79/pCDH-LUC-copGFP cells in BALB/c-nu mice, significant ocular lesions and proliferation of tumor cells were observed in the eyes. Meanwhile, Melphalan intervention significantly inhibited tumor cells in a dose-dependent manner, indicating that the mouse model of retinoblastoma was successfully constructed.