Retrovirus-mediated gene transfer of tumor necrosis factor alpha into colon carcinoma cells generates a growth inhibition.
Retrovirus-mediated gene transfer was employed to introduce two different cDNAs of the human tumor necrosis factor alpha gene (TNF) into colon carcinoma cell lines either sensitive (LoVo) or resistant (LS174T) to the external addition of TNF. The TNF variants differed in their 5'-sequences (HTNF: authentic TNF cDNA, SPTNF: TNF cDNA with a signal sequence of the IgG heavy chain). The integration of TNF-specific DNA into the genomic DNA, the expression of TNF-specific mRNA and the translation into TNF protein was shown by Southern-, Northern- and Western analysis with TNF-specific DNA- and RNA probes and anti-TNF-antibodies, respectively. Neomycin selected tumor cell clones expressed biologically active TNF up to 40 pg/ml. As shown in a growth assay, both variants of TNF caused an up to 80% growth inhibition in colon carcinoma cells. Furthermore, the cells constitutively producing TNF had a distinguishable morphological appearance and a reduced colony growth in soft agar compared to the parental cells. The subcutaneous injection of 1 x 10(6) TNF-expressing tumor cells into nude mice resulted in a reduced tumor progression in comparison to the controls.
Walther
W
W
Max-Delbruck-Center of Molecular Medicine, Berlin-Buch, Germany.
Fichtner
I
I
Uckert
W
W
D000818:Animals; D001483:Base Sequence; D002455:Cell Division; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D004351:Drug Resistance / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D009368:Neoplasm Transplantation; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000378:metabolism; D014410:Tumor Stem Cell Assay
D012333:RNA, Messenger; D012367:RNA, Viral; D014409:Tumor Necrosis Factor-alpha