一种表达增强型红色荧光蛋白的前列腺癌骨转移模型的建立

Establishment of a prostatic carcinoma osseous metastasis model expressing enhanced red fluorescent protein

目的：建立稳定表达增强型红色荧光蛋白（ERFP）的NOD／SCID-hu（非肥胖性糖尿病／重症联合免疫缺陷）小鼠前列腺癌骨转移模型。方法：选用6～8周龄体重为22～25g雄性NOD／SCID-hu小鼠，利用外科植入术将成人骨组织（HAB）植入小鼠皮下，3～4周后将携有ERFP基因的逆转录病毒载体PLXSN—DsRed2转染前列腺癌细胞PC3m与LNCaP，收获稳定表达ERFP的前列腺癌细胞PC-3m／ERFP与LNCaP／ERFP并配置成细胞悬液，通过小鼠尾静脉注入小鼠体内。6～8周后通过整体荧光显像系统观察小鼠体内移植入骨组织处转移前列腺癌细胞ERFP的表达。结果：2只小鼠死于术后第2天；10只存活小鼠中，4只在整体荧光显像系统观察下发出红色荧光，成功建立前列腺癌骨转移模型。结论：本方法建立的表达ERFP的NOD／SCID-hu小鼠前列腺癌骨转移模型稳定、可靠；实验结果可以在体外进行无创性动态观察，具较强的灵敏性及特异性，是进行前列腺癌骨转移基础及临床研究的理想模型。

Objective：To establish a stable prostatic carcinoma osseous metastasis NOD/SCID-hu mouse model expressing enhanced red fluorescent protein （ERFP）. Methods： A total of 12 male NOD/SCID-hu mice were selected to implant human adult bone （HAB） by surgical implantation subcutaneously. Then about 3～4 weeks later, human prostatic carcinoma cell lines PC-3m and LNCaP, transfected by retroviral vector PLXSN-DsRed2 containing ERFP gene, were harvested and injected into the NOD/SCID-hu mice through the lateral tail vein. Six to eight weeks after injection, all NOD/SCID-hu mice were observed under the whole-body fluorescent imaging system to look for the sites with the expression of ERFP. Results：Two mice succumbed the next day after surgical implantation. Enhanced red fluorescent was captured under the whole-body fluorescent imaging system in 4 mice of the 10 survived mice. The prostatic carcinoma osseous metastasis model was successfully established. Conclusions： The newly established prostatic carcinoma osseous metastasis model expressing enhanced red fluorescent protein （ERFP） is stable and reliable. The experiment results can be observed dynamically in vitro in a noninvasive way, with much higher sensitivity and specificity. The model is an optimal animal model to study the mechanisms of prostatic carcinoma osseous metastasis, suiting for both the basic and clinical research.