Research On The Effects And Molecular Mechanisms Of Resveratrol On The Reversal Of JQ1 Resistance In Human Chondrosarcoma

Objective:To investigate the reversal effect of resveratrol on human chondrosarcoma JQ1 resistant cells,and to explore its molecular mechanism.Methods:1.CCK8 was used to detect the effects of different concentrations of JQI on the growth of SW1353 cells and SW1353/JQ1-R cells,and calculated the drug resistance multiples.At the same time,the reversal of drug resistance of SW1353/JQ1-R cells by resveratrol was observed,and the reversal of drug resistance was calculated.2.Cell invasion assay was used to detect the invasion ability of SW1353 cells and SW1353/JQ1-R cells.At the same time,Western blot was used to detect the expression of Vimentin.3.The cell cloning method was used to detect the growth of SW1353 cells and SW1353/JQ1-R cells,and the cell growth of SW1353/JQ1-R cells after treated with resveratrol combined with JQ1 was also detected.4.The effect of resveratrol combined with JQ1 treatment on the apoptosis of SW1353/JQ1-R cells was examined using microphotographs.5.Cell counting and crystal violet staining were used to detect the effects of resveratrol combined with JQ1 treatment on the proliferation of SW1353/JQ1-R cells.6.Comet assay was used to detect the effect of resveratrol combined with JQ1 treatment on the DNA damage of SW1353/JQ1-R cells.7.Western blot was used to detect the expression of PI3K,p-AKT,p-mTOR,Bcl-XL and apoptosis-related protein Caspase3,DNA damage-related proteinγ-H2AX,and invasion-related protein Vimentin after treatment with resveratrol combined with JQ1.Results:1.The half inhibitory concentration(IC50)of JQ1 on SW1353 cells was 10.71±4.89 μmol/L,and the half inhibitory concentration of SW1353/JQ1-R cells was 43.36±5.44 μmol/L.The difference was statistically significant(***P<0.005),indicating that SW1353/JQ1-R cells showed obvious drug resistance characteristics,and the drug resistance fold was 4.052.Compared with SW1353 cells,W1353/JQ1-R cells show stronger invasion and proliferation ability,and have higher anti-apoptotic ability.3.The half inhibitory concentration(IC50)of JQ1 on SW1353/JQ1-R cells is 43.36±5.44 μmol/L μmol/L.The IC50 of Jcombination of JQ1 and 25 μmol/L Res on SW1353/JQ1-R cells is 7.46± 1.78 μmol/L.The combination of JQ1 and 25 μmol/L Res increased the sensitivity of SW1353/JQ1-R cells to JQ1,and the reversal factor of Res to SW1353/JQ1-R cells was 5.81.The difference was statistically significant(****P<0.001)4.Resveratrol combined with JQ1 treatment can promote apoptosis and DNA damage,inhibit cell proliferation and colony formation,but has no significant effect on cell invasion capacity.5.After resveratrol combined with JQ1 treatment,the expression levels of PI3K,p-AKT,and p-mTOR protein in SW1353/JQ1-R cells were significantly down-regulated(****P<0.001),and the expression level of anti-apoptotic proteins Bcl-XL was also significantly down-regulated(****P<0.001).However,the apoptosis-related proteins Caspase-3 and DAN injury-related proteins were increased(****P<0.001),while invasion-related protein Vimentin had no significant change.Conclusion:Resveratrol may play a role in reversing the resistance of human chondrosarcoma-resistant cells to JQ1 by promoting the apoptosis and DNA damage of resistant cells,inhibiting the expression of anti-apoptotic proteins in resistant cells,and inhibiting cell proliferation.This effect may be related to the inhibition of PI3K/Akt/mTOR signaling pathway.It may provide new solutions to the drug resistance in the treatment of chondrosarcoma.