Transforming growth factor-beta1 and renal cell cancer: cell growth, mRNA expression and protein production of cytokines.
OBJECTIVE
We evaluated the effects of transforming growth factor-beta1 (TGF-beta1) on growth activity, cytokine mRNA expression and cytokine protein production by renal cancer cells.


METHODS
Exogenous continuous exposure of biologically active TGF-beta1 was performed on ACHN cells at various concentrations of 0.1 to 30 ng./ml. and the number of cells was counted each culture day. To determine the index of S-phase cells the bromodeoxyuridine pulse labeling method was used. Reverse transcriptase-polymerase chain reaction (PCR) was done and PCR products were quantified. Each supernatant cytokine level was measured using an enzyme-linked immunosorbent assay.


RESULTS
TGF-beta1 significantly inhibited the growth activities of ACHN cells compared with controls. Bromodeoxyuridine labeling indexes after treating ACHN cells with TGF-beta1 (10 ng./ml.) showed that it began to decrease gradually after 24 hours and after 72 hours it was inhibited to approximately 40% compared with controls. The mRNA of TGF-beta1, TGF-beta types 1 and 2 receptors, interleukin-6 (IL-6) and granulocyte-macrophage colony-stimulating factor (GM-CSF) from ACHN cells was detected by reverse transcriptase-PCR assay. IL-6 and GM-CSF proteins were produced constitutively from ACHN cells but other cytokines were not. Adding TGF-beta1 to the cell culture medium did not influence mRNA expression, or the protein production of IL-6 or GM-CSF.


CONCLUSIONS
The inhibition of growth activities of ACHN cells by TGF-beta1 are mediated by its direct binding to specific receptors on ACHN cells followed by cell cycle inhibition, while TGF-beta1 seems to have no effect on the production of the cytokines studied.
Shimabukuro
Tomoyuki
T
Department of Urology, Ube Industries Central Hospital, Ube, Japan.
Ohmoto
Yasukazu
Y
Naito
Katsusuke
K
D002292:Carcinoma, Renal Cell / Q000378:metabolism* / Q000473:pathology*; D002455:Cell Division; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007680:Kidney Neoplasms / Q000378:metabolism* / Q000473:pathology*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis*; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000502:physiology*; D053773:Transforming Growth Factor beta1; D014407:Tumor Cells, Cultured
D016207:Cytokines; D012333:RNA, Messenger; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1