| Objective:To probe whether quercetin could inhibit the recurrence and metastasis after nude mice hepatocellular carcinoma model received curative operation.and to study the possible molecular mechanism of inhibiting recurrence and metastasis.Methods:1.HCCLM6 cell culture:HCCLM6 cells cultured with the high sugar DMEM containing 15% new fetal bovine serum in 37 ℃ and 5% CO2 incubator,mixed gas saturated humidity(95%).Once every 2 days in liquid.Single-layer adherent cell growth, to be full of the bottom of the bottle, with pancreatic protein digestive juice to digest into single cells, subculture; 2.the establishment of high metastasis nude mice model:Logarithmic phase growth of HCCLM6 cells were inoculated in nude mice right subaxillary, about 10d tumor,30 days to remove underarm tumor cut in 1mm3, which is orthotopic inoculated in the left hepatic of nude mouse.and remove the left liver of nude mice after ligation of the left liver;3.the experimental nude mouse were divided into control group (normal saline) and quercetin group (quercetin 75mg/kg d).Each group was intragastric administration once a day for five days every weeks, and 30 days total;4.mice were decapitated after medicined for 30 days.and observe the morphology of liver recurrent tumors stained with HE;5.Do western-blot to test the expression of P65, MMP-2 and E-Cadherin protein.Results:After the treatment of 30 days, all nude mice were killed, the tumor recurrence rate was 100%.The recurrence tumor volume of control group and quercetin group were 49.75 mm3 and 25.49 mm3, the difference was statistically significant(P<0.05), this showed that quercetin could inhibit the growth of tumor recurrence after liver cancer resection in nude mice.Cut the liver of nude mice layer by layer.count and measure the intrahepatic tumor metastasis.The liver metastasis rate was 100% in the control group of 5 nude mice.There were 4 mice have liver metastases in the quercetin group.the number of tumor metastasis in the control group is 2.20 ± 0.84,and 1.00± 0.71 in the quercetin group, The difference was statistically significant (P<0.05).that is say.quercetin can reduce the liver metastasis number of hepatocellular carcinoma after liver resection in nude mice.The Western blot were detect the liver tumor recurrence foci the expression of P65. MMP-2, E-Cadherin protein.Results show that, The expression of P65 protein in the liver tumor recurrence was 156.26+8.60 and 170.07+6.61 in the quercetin group and the control group, the difference was statistically significant (P<0.05).Compared with the control group, the expression of P65 protein in the quercetin group was significantly lower than that of the control group, and the expression of P65 protein was inhibited by quercetin.The expression of MMP-2 protein in the liver tumor recurrence was 169.21±5.32 å'Œ 158.17 ± 4.80 in the control group and the quercetin group.in the control group and the control group, the difference was statistically significant (P<0.05).The expression of MMP-2 protein in liver tumor tissues of the two groups was different, and the expression of quercetin group was lower than that in the control group.The expression of E-Cadherin protein in the liver tumor recurrence was 136.24±3.03å'Œ 144.70± 5.02 in the control group and the quercetin group.the difference was statistically significant (P<0.05).The expression of E-Cadherin protein was significantly different between the two groups in the nude mice, and the expression of the quercetin group was higher than that in the control group. The results showed that quercetin could up-regulate the expression of E-Cadherin protein.Conclusions:Quercetin can inhibit the growth of recurrence after liver cancer resection in nude mice and reduce the number of liver tumor spread.and it can inhibit the expression of P65 protein, up-regulate the expression of E-Cadherin protein and down regulate the expression of MMP-2 protein. |
PDF Full Text Request