稳定表达绿色荧光蛋白肝癌大鼠肿瘤微转移时间窗的筛选

Screen of time window of stable expression of enhanced green fluorescent for transplanted hepatocellular carcinoma in Buffalo rats

目的:探讨稳定表达增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)的肝癌Buffalo模型大鼠肿瘤微转移时间窗。方法:采用McA-RH7777/EGFP细胞株皮下注射成瘤后制备瘤块。取35只Buffalo大鼠,开腹直视下将瘤组织块种植到肝脏,制备大鼠肝癌模型。观察模型大鼠生物学行为,于种植后7、12、18、21、26d检测大鼠肝功能后分别处死7只大鼠,荧光显微镜下观察肝内肿瘤生长情况、肺转移情况。结果:种植后7d行磁共振成像(MRI)扫描见大鼠肝内均有肿瘤提示造模成功。种植后大鼠体质量7d内增长不明显,之后开始增加至21d达高峰,随后开始下降。大鼠肝内肿瘤体积7d时为(13.36±2.90)mm3,12d时为(162.5±69.71)mm3,26d时达(1 683.36±375.02)mm3。肝外转移情况:21d时肺部出现荧光显微镜下可见的小转移灶,26d时肺部出现肉眼可见多发小转移灶。肝功能检测发现26d时白蛋白水平较前明显下降(P<0.05)。结论:在稳定表达绿色荧光蛋白的肝癌Buffalo大鼠模型中,12~18d为肝癌的微转移期,21d后为晚期肝癌。

Objective：To discuss the time window of stable expression of enhanced green fluorescent for transplanted hepatocellular carcinoma in Buffalo rats.Methods：McA-RH7777/EGFP cells were subcutaneously implanted into the Buffalo rats＇ right thighs.Pieces of tumor tissue were implanted into the left lobes of the livers in 35 rats.The weights and serum albumin levels of rats were measured on day 7, 12, 18, 21, 26 after tumor implanted.The subsequent growth, metastasis of the implanted tumors were observed and evaluated with whole-body fluorescence optical imaging system, fluorescent microscopy.Results：The tumors were detected with MRI in all animals on day 7 after tumor implanted.There was no significant growth of weight within 7days, but increased significantly after and reached the peak value at day 21, then decreased.The mean tumor volumes was （13.36±2.90） mm^3 on day 7, （162.5±69.71） mm^3 on day 12, increased to （1 683.36±375.02） mm^3 on day 26 after tumor implanted.With the fluorescent microscopy, micrometastasis tumors can be detected in lungs on day 21 after tumor implanted.Multiple small lung metastases were detected on day 26.The albumin levels on day 26 after tumor implanted were significantly lower than before （P〈0.05）.Conclusions：The period from 12 to 18 days in the models may be classified as the micrometastasis stage, and it becomes advanced liver cancer after 21 days.