Effects Of Isoorientin On Hela Cells Apoptosis In Cervical Cancer

Objective: To investigate the effect of different concentrations of isoorientin on the proliferation of human cervical cancer hela cells in vitro and to investigate the regulation of PI3K/Akt/mTOR signaling pathway in hela cells by isoorientin.Methods: The hela cell line of human cervical cancer was used as the research object.Hela cells were treated with different concentrations of hesperidin(20umol/L,40umol/L,80umol/L,160umol/L)for 0h,24 h,48h,five replicate wells were set up for each concentration gradient.The proliferation inhibition rate of each group was detected by MTT method.The expression of PI3 K,Akt,P-Akt and mTOR in each group were detected by Western blotting.Results: The results of MTT assay showed that there was no significant difference(P>0.05)between the effect of drug and the negative control group when the different dose of isoorientin was applied to hela cells for 0h,there was no significant difference between each two different concentration groups(P>0.05).When the concentration of isoorientin was 20umol/L,the absorbance value began to decline,and the difference was statistically significant(P<0.05)compared with the negative control group.The absorbance values at 24 h and 48 h after drug exposure were significantly different from those at 0h after drug exposure(P<0.05),while the absorbance values at 24 h and 48 h after drug administration showed no significant difference(P>0.05).The absorbance decreased with the increase of drug concentration(P<0.05)when the concentration of isoorientin was less than 80umol/L.When the concentration of isoorientin was more than 80umol/L,the absorbance value was higher than that of drug concentration of 80umol/L(P<0.05).When the concentration of isoorientin was 80umol/Land the action time was 48 h,the absorbance value was the lowest.Cell proliferation inhibition rate(%)=(1-experimental OD value / negative experimental control OD value)× 100% according to the principle of MTT colorimetric method and the absorbance value.The inhibitory rates of different concentrations of isoorientin on hela cells at 0 h,24 h and 48 h were 0,0,0,0;14%,25.9%,46.6%,27.7%;23.5%,33.8%,61.1%,31.8% respectively.The results of Western blotting showed that different concentrations of isoorientin could inhibit the expression of p-Akt protein in hela cells 48 h,but had no effect on the expression of PI3 K,Akt,mTOR protein(P>0.05).When the concentration of isoorientin was ?80umol/L,the expression of p-Akt protein in hela cells decreased with the increase of drug concentration.When the concentration of isoorientin was> 80umol/L,the expression of p-Akt in hela cells increased with the increase of drug concentration(P<0.05)compared with the concentration of isoorientin at 80umol/L..Conclusions: 1.Isoorientin in a certain concentration range(?80umol/L)has an inhibitory effect on Hela cells in a concentration-and time-dependent manner.Inhibition of cervical cancer Hela cells was weakened beyond a certain drug concentration(>80umol/L).2.Isoorientin can down-regulate hela cells p-Akt protein expression,the mechanism of inhibition of isoorientin on cervical cancer hela cells may be through regulating the PI3 K / Akt signaling pathway.