The Biological Property Of Resveratrol On TPC-1 Cells

Objective:To observe the biological function of resveratrol?Res?on the human thyroid papillary carcinoma TPC-1 cells and its underlying molecular mechanisms.Methods:We analyzed the proliferation of Res on TPC-1 cells by CCK-8 at different time point and Transwell was performed to detect the change of cell migration.Flow cytometry?FCM?was used to detect the apoptosis rate and cycle of TPC-1 cells and then detect the mRNA expression of Bax/Bcl-2?Caspase 3 and Bcl in the TPC-1 cells by RT-PCR and its protein expression by Western blot,TPC-1 cells were treated with Res?0,75,100?mol/L?or 20?mol/L LY294002 and the ratio of Bax/Bcl-2 mRNA expression,the levels of Caspase 3 and Bcl of each group were detected by qRT-PCR and the protein expression of Akt,pAkt,Caspase3,Bcl-xl by Western blot?WB?.Results:1.Res?25,50?mol/L?can promote the proliferation of TPC-1 cells at 72 hours?P<0.05?,and Res?75-200?mol/L?can inhibit the proliferation of TPC-1 cells and had a concentration and time dependent effect?P<0.05?.2.Compared with the control,the migration ratio of TPC-1 cells were significantly reduced after TPC-1 cells treatment with 75 and 100?mol/L Res at 24,48 and 72h?P<0.05?.3.The G₀/G₁ phase was stagnated at 75 and 100?mol/L treated TPC-1 cells and the percentage of accumulation in the G₀/G₁ phase of TPC-1 cells were increased in a time-dependent manner.4.The TPC-1 cells apoptosis rate were elevated with increasing Res concentration in dosage and time dependent manner.5.The Bax/Bcl-2 ratio was increased in the 75 and 100?mol/L groups at each time period?P<0.05?,At 48 and 72h,the Bax/Bcl-2 ratio in the 100?mol/L group was higher than that in the 75?mol/L?P<0.05?;the Bax/Bcl-2 ratio was gradually decreased in a time-dependent at 75?mol/L?P<0.05?.6.Caspase3 mRNA expression was increased and Bcl-xl mRNA expression was decreased in the 75 and 100?mol/L groups at each time period?P<0.05?,Moreover,the expression level of Caspase3 mRNA in the 100?mol/L and was higher than that in the 75?mol/L at different checkpoint?P<0.05?;the mRNA expression levels of Caspase3 were gradually increased at 75,100?mol/L in a time-dependent manner?P<0.05?;However,for the of Bcl-xl mRNA expression,the 100?mol/L group showed a slightly decrease?P<0.05?.the mRNA expression of Bcl-xl at 48 and 72h was lower than that at 24h at 75?mol/L?P<0.05?.7.After 24h of Res treatment,the Caspase3 protein expression was increased at75?100?mol/L,However,the protein bands of Bcl-xl were significantly decreased at24h.After 48 and 72h of Res treatment,the caspase-3 protein bands of TPC-1 cell at100?mol/L group were gradually thickened and Bcl-xl protein bands were gradually decreased in a dose-dependent manner.8.After 48h of Res treatment,the mRNA expression of Caspase3 in each groups was gradually increased,the mRNA expression of Bcl-xl was gradually decreased,and the Bax/Bcl-2 ratio was gradually increased?P<0.05?.9.The expression of p-Akt?Bcl-xl were decreased in each group,by contrast the Caspase3 protein were increased.while addition with LY294002 were obviously different with Res group.Conclusion:1.Resveratrol may inhibit the proliferation and migration of TPC-1 cells and induce its apoptosis at concentration of 75?100?mol/L in vitro;2.Resveratrol may induces apoptosis of TPC-1 cells due to downregulating Bcl-xl,and upregulatingand caspase-3 expression via inhibiting PI3K/Akt signaling pathway activation.