Effects of meloxicam on vascular endothelial growth factor and angiopoietin-2 expression in colon carcinoma cell line HT-29

Abstract

To investigate the effect of meloxicam, a selected NSAIDs, on cell growth, expression of VEGF and angiopointin-2 (Ang-2) protein in HT-29 cell line, cultured HT-29 cells were treated with meloxicam of various concentrations for various lengths of time. The proliferation of HT-29 was detected by cell counting kit-8 (CCK-8), the cell cycle was determined by flow cytometer and the levels of VEGF and Ang-2 protein in supernatants were examined by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of VEGF and Ang-2 in cultured HT-29 were determined by real-time quantitative reverse-transcription polymerase chain reaction. Our results showed that treatment of meloxicam of different concentrations and for various lengths of time had a cytotoxicic effect on the cell proliferation of HT-29 cells in a concentration-dependant and time-dependant manner. Cell cycle analysis showed that the cells were mainly blocked in G0/G1 phase. The VEGF and Ang-2 protein levels in supernatants of the culture medium were decreased gradually in a concentration-dependent or time-dependent fashion. The mRNA expression of cox-2, VEGF and Ang-2 showed a gradual and concentration-dependent reduction. It is concluded that meloxicam can reduce the expression of VEGF and Ang-2 at the protein and mRNA level in colon carcinoma cell line.

Keywords

meloxicam / colon carcinoma cell line / vascular endothelial growth factor / angiopoietin-2

References

[1] McCollB. K., BaldwinM. E., RoufailS., et al.. Plasmin activates the lymphangiogenic growth factors VEGF-C and VEGF-D. J Exp Med, 2003, 198: 863-868

[2] PoltorakZ., CohenT., SivanR., et al.. VEGF145, a secreted vascular endothelial growth factor isoform that binds to extracellular matrix. J Biol Chem, 1997, 272: 7151-7158

[3] SaitoM., HamasakiM., ShibuyaM.. Induction of tube formation by angiopoietin-1 in endothelial cell/fibroblast co-culture is dependent on endogenous VEGF. Cancer Sci, 2003, 94990: 782-790

[4] ThurstonG., SuriC., SmithK., et al.. Leakage-resistant blood vessels in mice transgenically overexpressing angiopoietin-1. Science, 1999, 286: 2511-2514

[5] ThrustonG., RudgeJ. S., IoffeE., et al.. Angiopoietin-1 protects the adult vasculature against plasma leakage. Nat Med, 2000, 6: 460-463

[6] AhmadS. A., LiuW., JungY. D., et al.. Differential expression of angiopoietin-1 and angiopoietin-2 in colon carcinoma. A possible mechanism for the initiation of angiogenesis. Cancer, 2001, 92: 1138-1143

[7] HolashJ., MaisonpierreP. C., ComptonD., et al.. Vessel cooption, regression, and growth in tumors mediated by angiopoietins and VEGF. Science, 1999, 284: 1994-1998

[8] HolashJ., WiegandS. J., YancopoulosG. D.. New model of tumor angiogensis: dynamic balance between vessel regression and growth mediated by angiopoietins and VEGF. Oncogene, 1999, 18: 5356-5362

[9] LobovI. B., BrooksP. C., LangR. A.. Angiopoietin-2 displays VEGF-dependent modulation of capillary structure and endothelial cell survival in vivo. Proc Natl Acad Sci USA, 2002, 99: 11 205-11210

[10] SmithT. J.. Cyclooxygenase as the principal targets for the actions of NASAIDs. Rheum Dis Clin Nor Am, 1998, 24: 501-523

[11] SinicropeF. A., GillS.. Role of cyclooxygenase-2 in colorectal cancer. Cancer Metastasis Rev, 2004, 23: 63-75

[12] OshimaM., DinchukJ. E., KarmanS. L., et al.. Suppression of intestinal polyposis in Apc delta 716 knockout mice by inhibition of cyclooxygenase-2 (COX-2). Cell, 1996, 87: 803-809

[13] SenoH., OshimaM., IshikawaT. O., et al.. Cyclooxygenase-2 and prostaglandin E(2) receptor EP(2)-dependent angiogenesis in Apc (Delta716) mouse intestinal polyps. Cancer Res, 2002, 62: 506-511

[14] TomozawaS., TsunoN. H., SunamiE., et al.. Cyclooxygenase-2 overexpression correlates with tumor recurrence, especially haematogenous metastasis, of colorectal cancer. Br J Cancer, 2000, 83: 324-328

[15] WilliamsC. S., TsujiiM., ReeseJ., et al.. Host cyclooxygenase-2 modulates carcinoma growth. J Clin Invest, 2000, 105: 1589-1594

[16] DempkeW., RieC., GrotheyA., et al.. Cyclooxygenase-2: a novel target for cancer chemotherapy?. J Cancer Res Clin Oncol, 2001, 127: 411-417