Transforming growth factor-beta1 modulates metalloproteinase-2 and -9, nitric oxide, RhoA and alpha-smooth muscle actin expression in colon adenocarcinoma cells.
Colon carcinoma invasiveness is a process involving cell-cell and cell-matrix alterations, local proteolysis of the ECM (extracellular matrix) or changes in cytokine and growth factor levels. In order to evaluate the role of TGF-beta1 (transforming growth factor-beta1) and small G protein RhoA in tumour progression, the influence of TGF-beta1 treatment or RhoA-associated kinase inhibitor on the production of NO (nitric oxide) and MMP-2 and MMP-9 (metalloproteinases-2 and -9) was analysed in three human colon adenocarcinoma cell lines (HT29, LS180, SW948) representing different stages of tumour development. All the tested cell lines produced low amounts of MMP-2 and MMP-9. rhTGF-beta1 and the synthetic Rho kinase inhibitor (Y-27632) decreased MMP-2 secretion by colon cancer cells, especially in the most advanced stage of colon cancer. rhTGF-beta1 decreased NO secretion by cells, while Y-27632 had no effect on it. Immunoblotting with anti-RhoA antibodies followed by densitometry revealed that RhoA levels were slightly increased after incubation of colon carcinoma cells (SW948) with rhTGF-beta1. rhTGF-beta1 induced alpha-smooth muscle actin (alpha-SMA) expression, especially in high Duke's grade of colon cancer, while Y-27632 blocked it. Summing up, in colon carcinoma cells, TGF-beta1 and RhoA protein may regulate tumour invasiveness measured as MMP, NO and alpha-SMA expression or assayed using motility data and may be a good target for cancer therapy.
Paduch
Roman
R
Department of Virology and Immunology, Institute of Microbiology and Biotechnology, Maria Curie-Skłodowska University, Akademicka 19, 20-033 Lublin, Poland. rpaduch@poczta.umcs.lublin.pl
Kandefer-Szerszeń
Martyna
M
Szuster-Ciesielska
Agnieszka
A
Plewka
Krzysztof
K
D000199:Actins / Q000378:metabolism; D000230:Adenocarcinoma / Q000201:enzymology / Q000378:metabolism*; D000577:Amides / Q000494:pharmacology; D045744:Cell Line, Tumor; D002465:Cell Movement; D003110:Colonic Neoplasms / Q000201:enzymology / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000378:metabolism
D000199:Actins; D000577:Amides; D011725:Pyridines; D011994:Recombinant Proteins; D053773:Transforming Growth Factor beta1; C108830:Y 27632; D009569:Nitric Oxide; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D020742:rhoA GTP-Binding Protein