The Effect Of Desacetyluvaricin On The Expression Of TLR4 And P53 Protein In The HBV-related Hepatocellular Cancer Cell

ObjectBy researching the effect of Desacetyluvaricin on the expression of TLR4 and P53 protein in the HBV-related human hepatocellular cancer cell.To establish a fundamental base for putting Desacetyluvaricin into a new anticancer drug by disclosing its anticancer mechanism in molecular biologic method.Methods1 Observe the inhibitory action of Desacetyluvaricin on the HBV-related human hepato-cellular cancer cell line HepG2.2.15 by MTT assay and find out the correlation of dosage-effect and time-effect.2 FCM(flow cytometer) was used to analyze the expression of TLR4 and P53 protein of Desacetyluvaricin on the HBV-related human hepatocellular cancer cell line HepG2215.3 LCSM(laser confocal scanning microscopy) was used to observe the effect of Desacety-luvaricin on the expression of HBx protein in the HBV-related human hepatocellular cancer cell line HepG2.2.15.Result1 The HBV-related human hepatocellular cancer cell line HepG2.2.15 has been inhibited after treated by Desacetyluvaricin for 24h and 25μg/ml.The highest inhibitory rate of Desacety-luvaricin on HepG2.2.15 was 65%.It is obviously higher than 50%.2 The expression of TLR4 in control group and negative control group and DES group were 37.16%and 2.60%and 71.94%respectively,there were distinct difference in three group (P＜0.01).The expression of P53 in DES group and negative control group and control group were 32.6%and 12.3%and 3.3%respectively,there were distinct difference in three group (P＜0.05 or P＜0.01 ).3 In control group:cell nucleus volume was larger than normal cell.Nuclear membrane was obvious.HBx expressed uniformly in the cytoplasm.The fluorescence intensity of HBx was 51.08.In DES group:cellular volume decreases,membrane crinkles,Nuclear membrane wasn't obvious.The fluorescence intensity of HBx was 25.23.Comparison with the control group was distinct difference(P＜0.05). Conclusions1 Desacetyluvaricin had better inhibitory effect on the HBV-related human hepatocellular cancer cell line HepG2215 and the effect was concentration and time-dependent.2 Desacetyluvaricin induces the apoptosis of the HBV-related hepatocellular cancer cell may be related to upregulating the protein expression of P53.3 The mechanism of Desacetyluvaricin intervening of the HBV-related hepatocellular cancer cell is related to upregulating the protein expression of TLR4,to activating the inherent immune passageway and to improving the immune ability.4 HBx can proliferate the HBV-related hepatocellular cancer cell and inhibit the apoptosis of the hepatitis B virus related hepatocellular cancer cell.5 HBx protein decreases obviously after treated by Desacetyluvaricin on the HBV-related human hepatocellular cancer cell line HepG2215.