Dynamic Changes Of Shh Signaling Pathway During The Process Of GES-1 Cells Induced By Low Concentrations Of MNNG Into MC Cells.

Objective: To study the expression of Shh signaling pathway in the process of GES-1 cellsinduced by low concentrations of MNNG into MC cellsMethods: Taking the cryopreservation of GES-1 cells which were revived and passaged. Until80% cells mixing, the cells were placed into 6mm petri dishes after cell counting. Adding the0.01mol/L solution of MNNG into GES-1 cells which were growing strongly,the finalconcentration is 2×10^-5mol/L. Replacing the culture solution without MNNG after the cellscultured 24 hours without sunlight. The cells were collected at 0,3,7 days ,marked as MC0, MC3,MC7. The expression of ShhmRNA and Gli-1mRNA detected by RT-PCR. The products ofRT-PCR were doing agarose gel electrophoresis, analyzed by digital imaging systems.Thedifferent groups were compared by the relative intensity ratio of Shh,Gli-1/GAPDH. Theexperiment was repeated 3 times. The expression of Shh and Gli-1 protein detected by ICC, theresults were analyzed by Image-pro plus5.0 image analysis system.Result: No expression of Shh,Gli-1 mRNA and protein were detected in GES-1 cells.Expressionof Shh,Gli-1 mRNA and protein were detected in MC0,MC3,MC7 cells and graduallyincreasedConclusion: Shh signaling pathway involved in the process of GES-1 cells induced by lowconcentration of MNNG into MC cells, the expresstion of it gradually increased with time.