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Tumorigenicity of BALB3T3 A31 cells transfected with hamster‐complement‐C1s cDNA
Author(s) -
Sakai Norie,
Kusunoki Mayumi,
Nishida Munehiro,
Toyoguchi Toru,
Fukutomi Hisayuki,
Sakiyama Hisako
Publication year - 1994
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910580227
Subject(s) - transfection , microbiology and biotechnology , complementary dna , hamster , biology , cell culture , monoclonal antibody , fibroblast , antibody , immunology , gene , biochemistry , genetics
Hamster‐complement‐CIs cDNA was inserted into an expression plasmid BCMGSNeo (BCMGSNeoHACS). BALB/c mouse fibroblast A31 cells, which do not produce CIs, were transfected with BCMGSNeoHACS and the transfectants were selected with G418. Normal CIs production by the transfectants was confirmed by Northern and immunoblot analysis and by an esterase assay. To examine the tumorigenicity of the transfectants, 1 × 10 6 cells were injected s.c. into 6‐week‐old BALB/c nu/nu mice. Three CIs cDNA transfectants (A3CS9, A3CS12, A3CS13) formed tumors whereas both A31 and A31 transfected with the vector alone (A3BCMI and A3BCM3) did not. The tumors derived from the transfectants showed invasive growth, and many capillaries were observed in the tumors. A tumor derived from A3CS13 was examined immunohistochemically and found to be reactive with an anti‐CIs monoclonal antibody. Tumor cells were cultured in vitro again and CIs secreted into the culture medium was examined by immunoblot analysis. CIs synthesized by the tumor cells derived from A3CS13 maintained its biological functions. Tumor cells derived from A3CS9 and A3CS12 cells, however, produced CIs having abnormal disulfide bonds.