Kunzi-Rapp K, Genze F, Küfer R, Reich E, Hautmann R E, Gschwend J E
Department of Urology, University of Ulm, Ulm, Germany.
J Urol. 2001 Oct;166(4):1502-7. doi: 10.1016/s0022-5347(05)65820-x.
Chorioallantoic membranes have been used as a reliable biomedical assay system for many years. Chicken eggs in the early phase of breeding are between in vitro and in vivo systems but may provide an immunodeficient, vascularized test environment. We tested this model as an in vivo system for prostate cancer research.
Single cell suspensions of LNCaP, PC-3 and Tsu-Pr1 human prostatic cancer cell lines as well as 2 immortalized normal human prostate epithelial cell lines were inoculated on the chorioallantoic membrane of fertilized chicken eggs on day 5 or 6 of breeding. Tumor growth and viability of the embryo was evaluated by stereo microscopy. At day 10 the membranes were removed and embedded in paraffin. Cell morphology was assessed after hematoxylin and eosin staining. Cellular expression of cytokeratin, prostate specific antigen and androgen receptor as well as apoptosis induction was confirmed by immunohistochemistry.
Three days after tumor cell inoculation on the extraembryonic vascular system of the chorioallantoic membrane cell growth and formation of 3-dimensional tumors became apparent in 100% of inoculated membranes. Strong neo-angiogenesis was detected next to the established tumors and tumor cells invading the stroma of the chorioallantoic membrane. Cytokeratin expression as well as prostate specific antigen and androgen receptor in LNCaP cells confirmed the human prostate tumor origin. Assessment of quantitative in vivo apoptosis induction in LNCaP cells after intravenous injection of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate confirmed the model as a versatile in vivo system.
The well vascularized chorioallantoic membrane of bred chicken eggs is a suitable system for early in vivo cancer research. Reliable growth of prostate cancer cell lines is feasible and allows the evaluation of proliferation and apoptosis induction after intravascular or topic application of anticancer drugs. Exploitation of this assay enables a substantial reduction in or substitution for subsequent animal experiments.
多年来，绒毛尿囊膜一直被用作可靠的生物医学检测系统。处于繁殖早期的鸡胚介于体外和体内系统之间，但可能提供一个免疫缺陷的、有血管的测试环境。我们将该模型作为前列腺癌研究的体内系统进行了测试。
在繁殖第5天或第6天，将LNCaP、PC-3和Tsu-Pr1人前列腺癌细胞系以及2种永生化正常人前列腺上皮细胞系的单细胞悬液接种到受精鸡蛋的绒毛尿囊膜上。通过立体显微镜评估肿瘤生长和胚胎活力。在第10天，取出膜并嵌入石蜡。苏木精和伊红染色后评估细胞形态。通过免疫组织化学确认细胞角蛋白、前列腺特异性抗原和雄激素受体的细胞表达以及凋亡诱导情况。
在绒毛尿囊膜的胚外血管系统接种肿瘤细胞3天后，100%的接种膜中细胞生长和三维肿瘤形成明显可见。在已形成的肿瘤旁检测到强烈的新生血管生成，且肿瘤细胞侵入绒毛尿囊膜的基质。LNCaP细胞中细胞角蛋白表达以及前列腺特异性抗原和雄激素受体证实了人前列腺肿瘤起源。静脉注射佛波酯12-O-十四酰佛波醇-13-乙酸酯后，对LNCaP细胞体内定量凋亡诱导的评估证实该模型是一个通用的体内系统。
繁殖鸡蛋的血管丰富的绒毛尿囊膜是早期体内癌症研究的合适系统。前列腺癌细胞系可靠生长是可行的，并且允许在血管内或局部应用抗癌药物后评估增殖和凋亡诱导情况。利用该检测方法能够大幅减少或替代后续的动物实验。
