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Effect of MMC on Gene Expression and Distributing of NF-κB in SK-HEP-1 Cell
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Abstract:
Objective：To detect the effect of Mitomycin-C (MMC) on expression level and cellular distribution of NF-κB after MMC stimulation to further understand the mechanism of the NF-κB signal transduction in the response of SK-HEP-1 cell to the chemical drug. Methods：The cellular distribution and protein level of NF-κB was detected by Immunohistochemistry and Western blot method in SK-HEP-1 cell treated with different concentration of MMC for different duration. The mRNA expression of NF-κB gene was determined by RT-PCR in SK-HEP-1 cell treated with 50 μg/ml concentration MMC for different time after continuing treatment and 2 h treatment.Results：MMC stimulated the NF-κB influx into the nucleus from plasma and elevated the protein level of NF-κB and reached the peak at 8th hour. On effect of continual treatment of 50 μg/ml concentration MMC the expression level of NF-κB gene was elevated until the peak at 2nd h and then descended. However after treatment of MMC for 2 h and then withdrawal, the expression of NF-κB was elevated until 12th h. Conclusions：MMC could drive the NF-κB to come into the nucleus and elevate the expression of the gene. It was suggested that the NF-κB signal transduction pathway had relation to the response of SK-HEP-1 cell to MMC treatment.
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