The Effects And Mechanisms Of Inositol Hexaphosphate On Differentiation Of HT-29 Human Colon Carcinoma Cell Line

Objective: To investigate the effects and mechanisms of inositol hexaphosphate on differentiation of HT-29 human colon carcinoma cell line.Methods: Cells were exposed to various concentrations (0.0mM, 1.8mM, 3.3mM) of IP₆ for different time(1d,3d,5d). Its effects on the cells were evaluated by detecting follow indexes: 1. Observing ultrastructure of HT-29 cells by transmission electron microscopy. 2. To examine cell cycle by using flow cytometry. 3. C-myc mRNA expression state in cells were determined by reverse transcription polymerase chain reaction (RT-PCR). 4. Immunocytochemical stain was used to detect the expression of c-myc protein and Rb protein. 5. Detecting alkaline phosphate to evaluating cell differentiation.Results: 1.After treatment at 3.3mM concentration, cell structure changed, and appeared tendency of differentiation, such as karyopyknosis, abundant cellular organs and microvillus increasing. 2. IP₆ could block cell cycle and G0/G1 stage cells increasing, S stage and G2/M stage cells decreasing. 3. RT-PCR and the immunocytochemical stain showed that different concentration of IP₆ decreased the expression of c-myc mRNA and protein. 4. The immunocytochemical stain showed that that different concentration of IP₆ increased the expression of Rb protein. 4. The activity of alkaline phosphate increased with the concentration increasing and time going.Conclusion: The results show that IP₆ can induce differentiation of HT-29 cell, by augmenting the expression of Rb, decreasing the expression of c-myc and blocking cell cycle.