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The influence of TNF-α (tumor necrosis factor - alpha) on concentration of soluble adhesion molecules in cultures of HT-29 cells exposed to inositol hexaphosphate (IP6)

Beata Parfiniewicz ¹, Joanna Pendzich ², Małgorzata Kapral ¹, Ilona Bednarek ³, Ludmiła Węglarz ¹

1. Departament of Biochemistry, Medical University of Silesia, Narcyzow 1, Sosnowiec 41-200, Poland
2. The Third Uniwersity Hospital, Koziolka 1, Zabrze 41-800, Poland
3. Department of Biotechnology and Genetic Engineering, Narcyzów 1, Sosnowiec 41-200, Poland

Abstract

Continuous improvement in colon cancer therapy is one of the most difficult challenges of contemporary medicine, especially with regard to a high tumor metastatic potential, which is the main cause of treatment failure. The latest studies suggest that adhesion molecules are involved in the arising of malignant changes and in distant metastasis induction. The changes in their expression, and consequently, in the concentration of their soluble forms in serum, could be modulated by many different factors affecting cancer cells. In the case of colon cancer one of the factors is a high-fiber diet, containing an anti-cancer chemical, inositol hexaphosphate (IP6). It is known, however, that in both inflammatory diseases and the malignant tumor microenvironment the concentration of pro-inflammatory cytokines increases, including TNF-α. This cytokine can modify the adhesive properties of cells and influence the concentration of soluble adhesion molecules deriving from malignant colon cells. Nevertheless, there is no literature data on the synergistic effect of TNF-α and IP6 in the cancer cells microenvironment.

The goal of this study was to evaluate the influence of TNF-α on the concentration of soluble intercellular adhesive molecule-1 (sICAM-1) and soluble E-cadherin (sE-cadherin) in the microenvironment of HT-29 malignant cells stimulated with IP6. The elevated concentration of these soluble adhesive molecules suggests the intensification of the migration ability of cancer cells, and the concurrent increase in cancer cells invasiveness and metastasis.

HT-29 cell line derived from human colorectal cancer was used to evaluate sICAM-1 and sE-cadherin concentrations in their microenvironment. The cells were stimulated with 0.5 mM, 1.0 mM and 2.0 mM IP6, 10 ng/ml and 100 ng/ml TNF-α and TNF-α plus IP6 for 96 hours. Nonexposed cell line cultures as well as a culture medium without colon cancer cells served as controls. sICAM-1 and sE-cadherin concentrations were measured in the culture medium by immunoenzymatic method ELISA using Quantikine – Human sICAM-1/CD54 Immunoassay (R&D Systems) and Quantikine–Human sE-Cadherin Immunoassay (R&D Systems). All the results obtained were expressed as ng per ml.

The experiments showed the presence of both soluble ICAM-1 and soluble E-cadherin in HT-29 cell cultures exposed and unexposed to TNF-α and IP6. The control medium without the cells was negative for these adhesion molecules. Therefore, sICAM-1 and sE-cadherin were shed from HT-29 cells. After TNF-α treatment at concentration of 10 ng/ml of HT-29 cells, the values of sICAM-1 in cell culture supernatants did not change, but 100 ng/ml TNF-α caused a six-sevenfold increase of these soluble molecules compared with control culture. Interestingly, TNF-α at that concentration caused the increase in the sICAM-1 level, but to a lesser degree in the presence of higher concentrations of IP6. Similarly, as in the case of sICAM-1, TNF-α and IP6 caused dysregulation of sE-cadherin molecules in the intestinal epithelial of HT-29 cells. In our studies TNF-α caused an increase in sE-cadherin level in cultures exposed to 100 ng/ml of the cytokine, but 10ng/ml TNF-α did not influence their concentration in HT-29 cells cultures. A decrease in sE-cadherin in cultures supernatant under the influence of 2 mM IP6 was observed. The addition of TNF-α at a concentration of 10 ng/ml to the cultures containing 0.5, 1.0, and 2.0 mM IP6 did not cause any increase in the sE-cadherin level. Taking into account the concentrations of these molecules in the control culture, TNF-α at concentration of 100ng/ml did not significantly increase the sE-cadherin level in the presence of various IP6 concentrations either. Their level was most likely associated with IP6 activity. The higher concentration of IP6 and the lower sE-cadherin level in the presence of TNF-α was observed.

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The influence of TNF-α (tumor necrosis factor - alpha) on concentration of soluble adhesion molecules in cultures of HT-29 cells exposed to inositol hexaphosphate (IP6)

Presentation: Poster at VIII Multidyscyplinarna Konferencja Nauki o Leku, by Beata Parfiniewicz See On-line Journal of VIII Multidyscyplinarna Konferencja Nauki o Leku Submitted: 2012-03-15 11:43 Revised: 2012-03-15 11:43

Presentation: Poster at VIII Multidyscyplinarna Konferencja Nauki o Leku, by Beata Parfiniewicz
See On-line Journal of VIII Multidyscyplinarna Konferencja Nauki o Leku

Submitted: 2012-03-15 11:43 Revised: 2012-03-15 11:43