前列腺癌细胞株Du145裸鼠胫骨部骨肿瘤转移模型的建立

目的:建立一种裸鼠前列腺癌骨转移模型,观察肿瘤局部生长和远处转移情况,并进一步探讨其应用价值。方法:9只6周龄雄性BALB/C裸鼠麻醉后,将前肢及左后肢固定,以1 m l TB针筒-29G针头从胫骨头关节窝,刺入骨髓腔缓缓注入人前列腺癌Du145细胞悬液30μl(约5×106个细胞)至骨髓腔,以建立前列腺癌骨转移模型。观察裸鼠生命体征变化。于濒死状态下处死,取右后肢及其附近淋巴结、肺脏和肝脏等标本,用40 m l/L甲醛固定、石蜡包埋、苏木精-伊红染色后显微镜下观察。结果:9只裸鼠中有6只模型建立成功。接种后平均48 d裸鼠出现跛行现象,右后肢胫骨部可触及米粒大小的包块,质硬,进一步发展至小鼠行走障碍。55 d后出现恶病质,解剖后发现右后肢胫骨部骨质疏松,局部有“腐肉”样组织突出髓腔,填塞肌肉间隙,病理证实为肿瘤组织。肝脏泛黄、被膜皱缩,镜下呈急性重型肝炎的病理改变。结论:胫骨内注射细胞悬液制作裸鼠前列腺癌骨转移模型较好地模拟了人前列腺癌在骨骼微环境中的生长及转归情况,是研究前列腺肿瘤骨转移的适宜模型。

Establishment of Bone Metastasis Model of Prostate Cancer in Nude Mice by Intratibia Injection of Human Prostate Cancer Cell Line Du145

OBJECTIVE: To establish a bone metastasis model of prostate cancer by intratibia injection of Du145 in nude mice, observe the local growth of tumor in tibia and then assess application value of this model. METHODS: For 9 male nude mice, Du145 (5 x 10(6)) was injected in tibia by a TB syringe with a 29-gauge needle at a dose of 30 microl per mouse. Then the vital signs of the nude mice were observed. When the mice were dying, they were sacrificed, and the tissues of right hindlimbs, lymphatic nodes, lungs and livers were taken out, fixed in 10% formalin, embedded in paraffin, stained by HE and then observed microscopically. RESULTS: Incidence of bone tumor after intratibia injection was 67% (6 out of 9). About 48 days later, there were some small palpable nodes in right hind-limbs of the 6 mice and they couldn't walk normally. About 55 days later, cachexia occurred in them. After dissection, some carrion-like tissue grew from marrow cavity to muscular spatium, which was identified as tumor tissue by HE. The envelop of livers became crampy, and acute hepatitis could be diagnosed through microscopy, which represented a large scale of hepatocytic death, liver sinus dilatation and hyperemia, hepatic lobule infiltrated by lymphocyte, macrophage and inconspicuous hyperplasia. Since hypohepatia occurred too early, we couldn't detected distant metastases. CONCLUSION: The intratibia injection model is an optimal animal model to study metastasis of prostate cancer. It mimics the natural situation of human prostate cancer and will help to understand the mechanisms of androgen-independence and osseous metastasis, and tumor-host determinants of PSA expression.