Yang J, Liu A, Dougherty C, Chen X, Guzman R, Nandi S
Cancer Research Laboratory, University of California, Berkeley 94720, USA.
Anticancer Res. 2000 May-Jun;20(3A):1635-9.
Human tumor xenografts in nude mice are widely utilized model system for the transplantation of human surgical specimens and human established cell lines. Gene expression studies are often carried out in these model systems. With an increasing use of PCR based analyses, the extreme sensitivity of this technique poses a serious challenge with regards to the extent of contaminating host mouse cells in the human tumor xenografts. These xenografts are never free of host cell contamination. We detected mouse estrogen receptor expression in several human tumor xenografts using RT-PCR demonstrating that precaution is necessary when utilizing PCR based analyses in human tumor xenografts. A cytologically based methodology which distinguishes human versus mouse cells will be more suitable for ER expression studies using human xenograft models. Both (1) in situ hybridization using human probe and (2) immunocytochemistry using a monoclonal antibody directed against human cytokeratin have been used successfully to distinguish human cells versus host mouse cells in human xenografts in nude mice. Immunostaining of ER can then be utilized to determine the expression pattern of ER in the transplanted human cells.
裸鼠体内的人肿瘤异种移植是用于移植人类手术标本和已建立的人类细胞系的广泛使用的模型系统。基因表达研究经常在这些模型系统中进行。随着基于PCR分析的使用增加,该技术的极高灵敏度对人肿瘤异种移植中宿主小鼠细胞的污染程度提出了严峻挑战。这些异种移植永远无法摆脱宿主细胞污染。我们使用RT-PCR在几种人肿瘤异种移植中检测到小鼠雌激素受体表达,表明在人肿瘤异种移植中使用基于PCR的分析时必须采取预防措施。一种基于细胞学的区分人类细胞与小鼠细胞的方法将更适合使用人异种移植模型进行雌激素受体表达研究。(1)使用人探针的原位杂交和(2)使用针对人细胞角蛋白的单克隆抗体的免疫细胞化学已成功用于区分裸鼠人异种移植中的人类细胞与宿主小鼠细胞。然后可以利用雌激素受体的免疫染色来确定雌激素受体在移植的人类细胞中的表达模式。