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AIM: To investigate the possible involvement of 25-hydroxyvitamin D3-1α-hydroxylase [1α-25(OH)2D3] in butyrate-induced differentiation in human intestinal cell line Caco-2 cells. METHODS: Caco-2 cells were incubated either with 3 mmol/L butyrate and 1 μmol/L 25(OH)2D3 or with 1 μmol/L 1α-25(OH)2D3 for various time intervals ranging from 0 to 72 h. Additionally, cells were co-incubated with butyrate and either 25(OH)2D3 or 1α-25(OH)2D3. 1α-25(OH)2D3 mRNA was determined semi-quantitatively using the fluorescent dye PicoGreen. Immunoblotting was used for the detection of 1α-25(OH)2D3 protein. Finally, enzymatic activity was measured by ELISA. RESULTS: Both butyrate and 1α-25(OH)2D3 stimulated differentiation of Caco-2 cells after a 48 h incubation period, while 25(OH)2D3 had no impact on cell differentiation. Synergistic effects on differentiation were observed when cells were co-incubated with butyrate and vitamin D metabolite. Butyrate transiently upregulated 1α-25(OH)2D3 mRNA followed by a timely delayed protein upregulation. Coincidently, enzymatic activity was enhanced significantly. The induction of the enzyme allowed for comparable differentiating effects of both vitamin D metabolites. CONCLUSION: Our experimental data provide a further mechanism for the involvement of the vitamin D signaling pathway in colonic epithelial cell differentiation by butyrate. The enhancement of 1α-25(OH)2D3 followed by antiproliferative effects of the vitamin D prohormone in the Caco-2 cell line suggest that 25(OH)2D3 in combination with butyrate may offer a new therapeutic approach for the treatment of colon cancer.
AIM: To investigate the possible involvement of 25-hydroxyvitamin D3-1α-hydroxylase [1α-25 (OH) 2D3] in butyrate-induced differentiation in human intestinal cell line Caco-2 cells. METHODS: Caco- 3 mmol / L butyrate and 1 μmol / L 25 (OH) 2D3 or with 1 μmol / L 1α-25 (OH) 2D3 for various time intervals ranging from 0 to 72 h. 25 (OH) 2D3. 1α-25 (OH) 2D3 mRNA was determined semi-quantitatively using the fluorescent dye PicoGreen. Immunoblotting was used for the detection of 1α-25 (OH) 2D3 protein. , enzymatic activity was measured by ELISA. RESULTS: Both butyrate and 1α-25 (OH) 2D3 stimulated differentiation of Caco-2 cells after a 48 h incubation period while 25 (OH) 2D3 had no impact on cell differentiation. differentiation were observed when cells were co-incubated with butyrate and vitamin D metabolite. Butyrate transiently upregulated 1α-25 (OH) 2D3 mRNA followed by a timely delayed protein upregulation. Coincidently, enzymatic activity was significantly significantly. The induction of the enzyme allowed for comparable differentiating effects of both vitamin D metabolites. CONCLUSION: Our experimental data provide a further mechanism for the involvement of the vitamin D signaling pathway in enhancement of 1α-25 (OH) 2D3 followed by antiproliferative effects of the vitamin D prohormone in the Caco-2 cell line suggest that 25 (OH) 2D3 in combination with butyrate may offer a new therapeutic approach for the treatment of colon cancer.