人前列腺癌骨转移裸鼠移植模型的建立及其循环肿瘤细胞的特性  

Establishment of a nude mouse model of bone metastasis of human prostate cancer and the biological characteristics of circulating tumor cells

作　　者：张彩勤[1] 张海[1] 赵勇[1] 白冰[1] 刘佩娟[1] 师长宏[1] 

机构地区：[1]第四军医大学实验动物中心,西安710032

出　　处：《中国实验动物学报》2016年第1期14-19,共6页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金项目(31572340);军队实验动物专项课题(SYDW2014-002)

摘　　要：目的利用裸鼠移植模型研究雄激素阻断对人前列腺癌骨转移发生的影响,并初步探索肿瘤转移过程中循环肿瘤细胞(CTCs)的生物学特性。方法将40只裸鼠随机平分为两组,一组裸鼠手术切除睾丸以阻断雄激素的分泌,另一组为正常对照。心脏注射绿色荧光蛋白和荧光素酶双标的人前列腺癌细胞PC3-Luc-GFP,剂量为每只1×106/50μL,制备实验用骨转移模型。通过小动物活体成像系统检测人前列腺癌骨转移模型的建立;将发现转移的骨组织切片进行HE染色分析,并分离转移模型中CTCs,通过Western blot检测CTC中转移相关分子RANKL和TOPK的表达。结果成功建立了前列腺癌骨转移模型,切除睾丸组的骨转移发生率为2/13(15.38%),而正常对照组的骨转移发生率为5/14(35.71%);再次重复裸鼠移植模型的建立,合并两次动物实验结果得出:切除睾丸组的骨转移发生率为3/26(11.54%),而正常对照组的骨转移发生率为10/27(37.04%),两组间差异有显著性(P<0.05);CTCs中RANKL和TOPK表达量均明显高于原始的前列腺癌细胞。结论在裸鼠移植模型中阻断雄激素能显著降低前列腺癌骨转移的发生率。CTC的转移能力高于原始的前列腺癌细胞。Objective To study the influence of androgen deprivation on prostate cancer bone metastasis in nude mouse models and to explore the biological characteristics of circulating tumor cells during tumor metastasis. Methods Forty male 6-7-week old nude mice were randomly divided into two groups： the castration group and untreated normal control group. All the nude mice were injected intracardially human prostate cancer PC3-Luc-GFPcells（ labeled with GFP and Luc） in a dose of 1 × 106/50 μL / mouse to establish the bone metastasis models. The resulted cancer bone metastasis was confirmed by detection of Luc signal using a small animal optical imaging system. Bone tissue samples were fixed with 4%paraformaldehyde and pathological examination using HE staining was performed. Circulating tumor cells（ CTCs） were isolated from the periphery blood collected by heart puncture. The expression of metastasis-associated molecules TOPK and RANKL of CTCs was detected by Western blot assay. Results The nude mouse model of human prostate cancer bone metastasis was successfully established. The incidence rate of bone metastasis in the castration group was 2 /13（ 15. 38%）,while that of the normal control group was 5 /14 = 35. 71%. The xenograft models were prepared twice and the results of the two experiments were pooled together,showing a bone metastasis incidence rate of 11. 54%（ 3 /26） in the castration group and 37. 04%（ 10 /27） in the normal control group（ P〈0. 05）. The expressions of TOPK and RANKL of CTCs were significantly higher than that in the original prostate cancer cells. Conclusions Castration can significantly reduce the incidence of prostate cancer bone metastases in nude mouse models. Metastatic potential of CTCs is significantly higher than that of the original prostate cancer cells.

关 键 词：前列腺癌 骨转移模型 裸鼠模型 睾丸切除 循环肿瘤细胞 

分 类 号：Q95-33[生物学—动物学]