| Objective: To investigate the relationship between GDNF mRNA expression andhistone acetylation of the promoter regions in rat C6glioma cells.Methods: RT-PCR、Real-time PCR and ChIP-PCR techniques were used respectivelyto compare the expression level of GDNF mRNA and acetylation status of histone H3K9inpromoter regions Ⅰ and Ⅱ in rat normal astrocytes and C6glioma cells; After treatingwith varying concentrations of histone acetyltransferase inhibitor Curcumin or deacetylaseinhibitor trichostatin A (TSA) in C6glioma cells, the mRNA expression and histone H3K9acetylation in Promoter Regions I and II of the GDNF gene was detected by real-time PCRand ChIP-PCR.Results: Compared with that in normal astrocytes, the GDNF mRNA expressionincreased significantly in C6glioma cells (p<0.01), and the acetylation of histone H3K9also enhanced in promoter regions Ⅰ and Ⅱ significantly (P<0.05). At24h after treatmentwith Curcumin, the expression of GDNF mRNA decreased in concentration-dependentmanner in C6glioma cells, and about74.17%decreased was observed in100μM group (P<0.001); In contrary, GDNF mRNA expression rased after TSA treatment, and anear-maximum increase of145.35%was observed in200nM group (P <0.05). Meanwhile,the acetylation level of histone H3K9in promoter region Ⅱ of GDNF gene was sensitiveto Curcumin or TSA treatment, where the concentration-course for acetylation was similarto that for gene transcription.Conclusion: The level of histone H3K9acetylation in GDNF promoter regions Ⅰ and Ⅱincreased significantly in glioma cells; Hyperacetylation of histone H3K9in the promoterregion Ⅱ may be the cause of aberrantly high transcription of the GDNF gene in gliomacells. |
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