Tumorigenicity of BALB3T3 A31 cells transfected with hamster-complement-C1s cDNA

✍ Scribed by Norie Sakai; Mayumi Kusunoki; Munehiro Nishida; Toru Toyoguchi; Hisayuki Fukutomi; Hisako Sakiyama

• Book ID: 102866934
• Publisher: John Wiley and Sons
• Year: 1994
• Tongue: French
• Weight: 633 KB
• Volume: 58
• Category: Article
• ISSN: 0020-7136
• DOI: 10.1002/ijc.2910580227

✦ Synopsis

Hamster-complement-C I s cDNA was inserted into an expression plasmid BCMGSNeo (BCMGSNeoHACS). BALBIc 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, I x lo6 cells were injected S.C. into 6-week-old BALBIc nulnu mice. Three C I S cDNA transfectants (A3CS9, A3CS 12, A3CS 13) formed tumors whereas both A3 I and A3 I 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 A3CS I3 was examined immunohistochemically and found to be reactive with an anti-CIS monoclonal antibody. Tumor cells were cultured in vitro again and C I S secreted into the culture medium was examined by immunoblot analysis. C I s synthesized by the tumor cells derived from A3CS I3 maintained its biological functions. Tumor cells derived from A3CS9 and A3CS I 2 cells, however, produced C I s having abnormal disulfide bonds.