Ananthaswamy H N, Price J E, Goldberg L H, Bales E S
Department of Immunology, University of Texas M.D. Anderson Hospital and Tumor Institute, Houston 77030.
J Cell Biochem. 1988 Feb;36(2):137-46. doi: 10.1002/jcb.240360205.
High-molecular-weight genomic DNA isolated from a human cutaneous squamous cell carcinoma (AS) was assayed for its ability to induce tumorigenic transformation of NIH 3T3 cells. Subcutaneous injection of NIH 3T3 cells cotransfected with DNAs from AS tumor and pSV2-neo plasmid not only induced tumors at the site of injection, but also metastasized spontaneously to the lungs in 100% of nude mice injected. DNA isolated from a representative primary tumor and a metastasis was again used in a second round of transfection. Injection of secondary transfectants into nude mice again resulted in induction of both subcutaneous tumors and spontaneous long metastases. Southern blot hybridization with ras-specific probes revealed that DNA from both primary tumors and metastases induced by AS tumor DNA contained highly amplified Ha-ras oncogene. Furthermore, DNAs from secondary tumors and metastases induced by DNA from a primary tumor and a metastasis also contained similar highly amplified Ha-ras oncogene. These results suggest that the amplified Ha-ras oncogene may be responsible for induction of both tumorigenic and metastatic phenotypes in NIH 3T3 cells transfected with DNA from AS tumor.
对从人皮肤鳞状细胞癌（AS）中分离得到的高分子量基因组DNA进行检测，以评估其诱导NIH 3T3细胞发生致瘤转化的能力。皮下注射与来自AS肿瘤的DNA和pSV2-neo质粒共转染的NIH 3T3细胞，不仅在注射部位诱导形成肿瘤，而且在100%注射的裸鼠中自发转移至肺部。从一个代表性的原发性肿瘤和一个转移瘤中分离得到的DNA再次用于第二轮转染。将二次转染细胞注射到裸鼠体内再次导致皮下肿瘤和自发性远处转移的形成。用ras特异性探针进行Southern印迹杂交显示，由AS肿瘤DNA诱导的原发性肿瘤和转移瘤的DNA均含有高度扩增的Ha-ras癌基因。此外，由原发性肿瘤和转移瘤的DNA诱导的二次肿瘤和转移瘤的DNA也含有相似的高度扩增的Ha-ras癌基因。这些结果表明，扩增的Ha-ras癌基因可能是导致用来自AS肿瘤的DNA转染的NIH 3T3细胞产生致瘤和转移表型的原因。
