A Research On The Effects Of Resveratrol On The Nucleolar Function And Its Molecular Mechanism

The purpose of the studyInvestigate of the effects of resveratrol on the nucleolar function and how to influence the nucleolar functions through PI3K/Akt signal pathway preliminarily.Methods and results1. Through the MTT, the clone formation and DAPI staining technique for the detection of the effect of resveratrol on the proliferation rates, colony formation and cell apoptosis of HeLa cells. The results show that 24h after Res treament of 100μmol/L inhibit the proliferation and colony formation of HeLa cells, and restrain the proliferation and cloning formation of HeLa cells completely after 24h of Res treament of 400μmol/L and above 400μmol/L.2. Through the Luciferase Assay technology, CHIP technology and Realtime PCR technology for detection of the effect of Res on the rDNA promoter activity、HeLa cell transcription factor in rDNA promoter on the enrichment factor and pre-rRNA expression of HeLa cells, the results showed that:3h after Res was given, rDNA promoter activity of HeLa cells was inhabited; 12h after Res was given, the enrichment of RPA194 which is the subunit of RNA polymerase I and transcription factor UBF in rDNA promoter decreased gradually; 12h after Res was given, the expression of pre-rRNA after the relative amount of not adding drug group inhibition rate was about 70%, and in a concentration and time rely on the trend.3. Through Western Blotting technology for the detection Res’ effect of the expression of the main members activity of the PI3K/Akt signaling pathway, the results showed that:12h after Res was given, with the increase of concentration, the expression of UBF and Akt, which are the main members of PI3K/Akt signaling pathway, had no significant change, but the level of phosphorylation of UBF and Akt was significantly decreased, and in a concentration and time trend; compared with the inhibitor PI3Kα、PI3Kβ and Ly294002, the inhibitory effects of the inhibitor PI3K a was best, it shows that the subunit of PI3Kα played a major role in PI3K/Akt signaling pathway.4. Through the Luciferase Assay technology, CHIP technology, Realtime PCR technology and Western Blotting technology for detection of Res’effect on HeLa cells transfected plasmid of PIK3CA, the results showed that:Res can inhibit the activity of rDNA promoter of HeLa cells which PI3K/Akt signal pathway was activitied, reducing the activity of RPA194 which is the subunit of RNA polymerase I, inhibiting the expression of pre-rRNA, inhibiting the phosphorylation of UBF and Akt.ConclusionResveratrol is able to inhibit the proliferation of HeLa cells and promote the apoptosis of HeLa cells, lower the activity of RPA194 which is the subunit of RNA polymerase I, decreased the activity of rDNA promoter, down regulated the expression of pre-rRNA, decreased the phosphorylation of UBF and Akt in the PI3K/Akt signaling pathway, also has the obvious inhibitory effect on the PI3K pathway which is actived completely.