Department of Biological Sciences, National University of Singapore, Singapore, Singapore 117543.
Mar Biotechnol (NY). 2010 Nov;12(6):640-9. doi: 10.1007/s10126-009-9251-9. Epub 2010 Jan 6.
A retrovirus homologue gene of cellular cyclin D₁, walleye dermal sarcoma virus rv-cyclin gene (orf A or rv-cyclin), was expressed in the livers of zebrafish under the control of liver fatty acid-binding protein (lfabp) promoter. To prevent possible fatality caused by overexpression of the oncogene, the GAL4/upstream activation sequence (GAL4/UAS) system was used to maintain the transgenic lines. Thus, both GAL4-activator [Tg(lfabp:GAL4)] and UAS-effector [Tg(UAS:rvcyclin)] lines were generated, and the rv-cyclin gene was activated in the liver after crossing these two lines. Since no obvious neoplasia phenotypes were observed in the double-transgenic line, cancer susceptibility of the transgenic fish expressing rv-cyclin was tested by carcinogen treatment. Unexpectedly, transgenic fish expressing rv-cyclin gene (rvcyclin+) were more resistant to the carcinogen than siblings not expressing this gene (rvcyclin-). Lower incidences of multiple and malignant liver tumors were observed in rvcyclin+ than in rvcyclin- fish, and the liver tumors in the rvcyclin+ group appeared later and were less malignant. These results suggest that expression of rv-cyclin protects the fish liver from carcinogen damage and delays onset of malignancy. These findings indicate that transgenic fish models are powerful systems for investigating mechanisms of inhibition and regression of liver tumors.
细胞周期蛋白 D₁的逆转录病毒同源基因、黄鲈皮肤肉瘤病毒 rv-cyclin 基因(orf A 或 rv-cyclin),在肝脏脂肪酸结合蛋白(lfabp)启动子的控制下在斑马鱼肝脏中表达。为了防止癌基因过表达可能导致的致命性,使用 GAL4/上游激活序列(GAL4/UAS)系统来维持转基因系。因此,产生了 GAL4-激活子[Tg(lfabp:GAL4)]和 UAS-效应子[Tg(UAS:rvcyclin)]系,并且在这两条线杂交后,rv-cyclin 基因在肝脏中被激活。由于在双转基因系中未观察到明显的肿瘤表型,因此通过致癌物处理测试了表达 rv-cyclin 的转基因鱼的癌症易感性。出乎意料的是,表达 rv-cyclin 基因(rvcyclin+)的转基因鱼对致癌物的抵抗力强于不表达该基因(rvcyclin-)的同窝鱼。在 rvcyclin+鱼中观察到的多发性和恶性肝肿瘤的发生率低于 rvcyclin-鱼,并且 rvcyclin+组的肝肿瘤出现较晚且恶性程度较低。这些结果表明,rv-cyclin 的表达保护鱼的肝脏免受致癌物的损害,并延迟恶性肿瘤的发生。这些发现表明,转基因鱼模型是研究肝肿瘤抑制和消退机制的强大系统。