非肥胖糖尿病／严重联合免疫缺陷小鼠肾包膜下患者来源前列腺癌异种移植模型的建立

Establishment of subrenal capsule xenograft models of patient-derived human prostate cancer in nonobese diabetic/severe combined immunodeficiency mice

目的建立非肥胖糖尿病／严重联合免疫缺陷（nonobese diabetic／severe combined immunodeficiency，NOD／SCID）小鼠肾包膜下患者来源的前列腺癌异种移植模型，为前列腺癌病因学研究和个体化药物治疗提供理想的T具。方法2012年10月至2013年8月选取20只NOD／SCID小鼠。4～6周龄。体质量28～30g。采用完全随机分组法分为4组，每组5只，分别将4例前列腺癌根治术后新鲜癌组织经显微外科手术移植到NOD／SCID小鼠肾包膜下，每组对应1例前列腺癌组织。移植术后8～12周，在麻醉状态下，解剖小鼠肾脏，观察肿瘤生长情况。切除成瘤肾脏，采用HE染色检查进行组织学诊断。采用兔抗人抗体进行免疫组化染色检测PSA、α-甲酰基辅酶A消旋酶（α-methylacyl CoA racemase，AMACR／P504S）和细胞核中P63蛋白的表达情况。结果前列腺癌组织在NOD／SCID小鼠肾包膜下成活率为95％（19／20）。成活的前列腺癌组织为实体状、白色、隆出肾脏表面，伴有血管生成。镜下表现：腺体结构紊乱或被癌细胞破坏，前列腺癌细胞增殖旺盛，细胞核浓染，或呈多形性，可见异常分裂象。免疫组化染色检测示P504S阴性，P63显示腺腔基底膜消失或破坏，PSA强阳性，证实为人源性前列腺癌组织。结论采用显微外科技术在NOD／SCID小鼠肾包膜下建立患者来源的前列腺癌异种移植模型具有可靠的成活率。该模型较完整地保存了前列腺癌的异质性，再现了前列腺癌细胞亚群的生物学特性，是可靠的动物模型。

Objective To establish the xenograft model of human prostate cancer（PCa） by grafting patient-derived tissues beneath the renal capsule of intact male non-obese diabetic/severe combined immunodeficiency （NOD/SCID） mice. Methods Between October 2012 and August 2013, twenty NOD/SCID mice were randomly divided into 4 groups （n = 5 each）. Four patient-derived PCa specimens were collected and sent for frozen section analysis. A specimen of one patient corresponds to 5 mice. At 8-12 week after initial microsurgical implantation,grafts were harvested and fixed to be embedded with paraffin. For histopathology, routine bematoxylin-eosin staining was performed on these formalin-fixed and paraffin-embedded sections. Immunohistochemical studies were performed for the expression of α-methylacyl CoA racemase （AMACR/P504S, prostate-specific antigen and P63 antigen on the sections. Results Taken rate of prostate cancer xenogtrafts in subrenal capsular was 95% （19/20）. The subrenal capsule xenografts are protruded the renal surface and embedded into the renal parenchyma with angiogenesis. Pathological sections showed disruption of the basal cell. There was significant increase in proliferation and anaplasia of malignant prostate epithelium cells at subrenal capsule xenografts. Xenografts were confirmed as human PCa tissues with the PSA （ ＋）/P63 （-）/P504S （-） immunostain. Conclusions The patient-derived human prostate cancer xenograft model in NOD/SCID mice is successfully established by microsurgical technique. This model can keep the heterogeneity of prostate, which could demonstrated the characters of PCa cells. It has been shown to facilitate the investigation of etiology mechanism of PCa, new agents and individualized antineoplastie therapy.