O'Hayer Kevin M, Counter Christopher M
Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, North Carolina, USA.
Methods Enzymol. 2006;407:637-47. doi: 10.1016/S0076-6879(05)07050-3.
Transgenic mice, cultured murine cells, and human cancer cell lines have widely been used to study Ras oncogenesis. Although extremely valuable systems, they could not be used to study Ras function in genetically defined human cells. In this regard, Ras is required for tumor formation in normal human somatic cells expressing SV-40 T/t antigens, which inactivate the tumor suppressors p53 and Rb and activate the oncogene c-Myc, and hTERT, the catalytic subunit of telomerase. Such a system allows not only the general requirements of Ras to be dissected in matched cells from different organisms or tissues but also the individual pathways required for tumor growth to be defined in human cells. This review will detail the methods of creating stable T/t Ag, TERT, Ras-expressing cell lines, as well as commonly used techniques of soft agar and xenograft tumor formation.
转基因小鼠、培养的鼠细胞和人类癌细胞系已被广泛用于研究Ras致癌作用。尽管它们是极有价值的系统，但无法用于研究Ras在基因定义的人类细胞中的功能。在这方面，在表达SV-40 T/t抗原的正常人类体细胞中，Ras是肿瘤形成所必需的，SV-40 T/t抗原会使肿瘤抑制因子p53和Rb失活，并激活癌基因c-Myc以及端粒酶的催化亚基hTERT。这样的系统不仅能在来自不同生物体或组织的匹配细胞中剖析Ras的一般需求，还能在人类细胞中确定肿瘤生长所需的各个途径。本综述将详细介绍创建稳定表达T/t抗原、TERT和Ras的细胞系的方法，以及常用的软琼脂和异种移植肿瘤形成技术。
