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Simultaneous transfer of tumorigenic and metastatic phenotypes by transfection with genomic DNA from a human cutaneous squamous cell carcinoma
Author(s) -
Ananthaswamy Honnavara N.,
Price Janet E.,
Goldberg Leonard H.,
Bales Elise S.
Publication year - 1988
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240360205
Subject(s) - transfection , oncogene , cancer research , metastasis , southern blot , biology , dna , microbiology and biotechnology , 3t3 cells , cell , carcinogenesis , cell culture , cancer , pathology , medicine , cell cycle , genetics
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.