人肾细胞癌原位移植裸鼠模型的建立及转移相关基因表达分析

Establishment of orthotopic mouse model of human renal cell carcinoma and expression analysis of metastasis-associated genes

目的:建立原位移植裸鼠模型,筛选同一标本来源的转移性和非转移性肾细胞癌(RCC)。方法:采用皮下移植法、细胞悬液原位注射法、肾周筋膜内法、组织块原位移植肾包膜下法对BALB/c裸小鼠接种(或注射)RCC组织块(或细胞悬液),观察各种方法的成瘤及转移情况,并采用免疫组化方法初步对比分析了VEGF、bFGF、P16、Bc l-2、C-m et在裸小鼠体内的转移性和非转移RCC的表达情况。结果:(1)组织块原位移植肾包膜下法成瘤率及转移率最高,分别为73.3%(11/15)、20%(3/15)。(2)与原发灶相比,转移灶中VEGF表达明显升高(P<0.05),C-m et表达明显降低(P<0.05),bFGF、Bc l-2、P16表达降低但无统计学差异(P>0.05)。结论:组织块原位移植肾包膜下法是切实有效的RCC转移模型制作方法。采用该法在裸鼠中获得了人转移性和非转移RCC,免疫组化分析证实了转移性RCC中新血管生成因子VEGF表达显著增加。

Objective：To establish orthotopic mouse models of human renal cell carcinoma and to separate metastatic or non-metastatic renal cell carcinoma （RCC） from the same source. Methods： Surgical specimens/cell suspensions were transplanted into the various tissues of BALB/c nude mice （subcutis, cellular orthotopic injection into renal capsule, perinephrium, surgical orthotopic implantation into renal capsule）. Tumorigenicity and metastasis were subsequently evaluated. Immunohistochemical analysis was employed to determine the expression of VEGF, bFGF, P16, Bcl-2 and C-met in the metastatic and non-metastatic RCC xenograft. Results： The incidences of tumorigenicity and metastasis of orthotopic model were the highest,being at 73.3% （11/15）and 20%（3/15）, respectively. Compared with the non-metastastic RCC, VEGF expression was upregulated in the metastatic RCC（ P〈0.05）, while the expression of C-met was downregulated significantly（P〈0.05） ,the expression of bFGF, Bcl- 2 and P16 was also downregulated but without significance. Conclusion： Mouse orthotopic transplantation is the most effective way for the growth and natural metastasis of human RCC. Primary and metastatic RCC from the same patient has been successfully harvested. Expression of VEGF in the metastatic tumor tissues is significantly higher than that in the non-metastatic RCC.