Expression Of C-Met Protein In Vitro Hyperthermia Residual Cancer Cell Model Of Hepatocellular Carcinoma After Radiofrequency Ablation

Objective:c-Met protein is closely related to the growth,proliferation,migration,invasion and apoptosis of tumor cells.In this study,we established the HepG2 hyperthermia model in vitro to simulate the residual cancer cells after radiofrequency ablation of hepatocellular carcinoma.The expression of c-Met protein in residual cancer cells was detected by immunocytochemistry.The aim of this study was to investigate the effect of hyperthermia on c-Met protein expression in residual cancer cells and clear c-Met protein on the role of residual cancer cells.And to provide the theoretical basis and experimental basis for the mechanism of recurrence and metastasis of HCC after RFA and the drug research with c-Met as the target.Methods:1?HepG2 cells were cultured at a normal concentration of 210 m L / L oxygen,50 g /L carbon dioxide,saturated humidity and 37°C in normal incubators.The liquid culture medium was DMEM cell culture medium containing 10% fetal bovine serum,and the culture medium was replaced every other day.Human hepatocellular carcinoma cell line HepG2 was adherent.The cells used in the experiment were in logarithmic growth phase.2?The establishment of experimental group in vitro thermotherapy residual cancer cell model: The logarithmic growth phase of HepG2 cells were placed in a cell incubatorcontaining normal concentrations of 210 mL / L of oxygen,50 g / L of carbon dioxide and saturated humidity.The culture medium was DMEM cell culture medium containing10% fetal bovine serum.The temperature of the incubator was adjusted to 41 ?,45 ?and 50 ? respectively.According to the different culture temperature,then divided into A?B?C three groups?The growth of HepG2 cells was observed at 6 hs,12 hs,24 hs and48 hs,and the residual cancer cells were subjected to subsequent experiments.3?A control group of residual cancer cell model: The logarithmic growth phase of HePG2 cells was placed in a cell incubator containing normal concentrations of 210 mL /L of oxygen,50 g / L of carbon dioxide and saturated humidity.The culture medium was DMEM cell culture medium containing 10% fetal bovine serum.The temperature of the incubator was maintained at 37 ° C,the growth of HePG2 cells was observed at 6 hs,12 hs,24 hs and 48 hs,and the residual cancer cells were subjected to subsequent experiments.4?Immunocytochemical method was used to detect the expression of c-Met protein HepG2 residual cancer cells in each group.5?SPSS software statistical analysis:(1)The difference of the positive rate of c-Met protein expression between the two groups of the experimental group A,B and C is statistically analyzed by ? 2 test.(2)The ? 2 test was used to compare the difference of the positive rate of c-Met protein expression between the experimental group and the control group(P <0.05).Results:1?The positive cells of c-Met protein were stained brown cytomembrane and the cell nucleus were not stained.2?The positive rates of c-Met protein expression in vitro hyperthermia residual cancer cell model of experimental group A,B and C were 33.2%,35.7% and 36.5%respectively.Although the positive rates of three were increased,but A and B,B and C,between A and C are compared,the differences were not statistically significant(PAB=0.144>0.05,PBC=0.648 > 0.05,PAC=0.055 > 0.05).The results showed that when the hyperthermia temperature is higher than a certain value(such as 37 ?),the positive expression rate of c-Met protein did not increase with the increase of temperature,and it is not dependent on hyperthermia temperature.3?The positive rates of c-Met protein expression in vitro hyperthermia residual cancer cell model of experimental group A,B and C are 33.2%,35.7% and 36.5%respectively,the positive rate of c-Met protein expression in the control group is 10.5%.A compared with the control group was statistically significant(P<0.001),B compared with the control group was statistically significant(P<0.001),C compared with the control group was statistically significant(P<0.001).Compared with the control group,the positive rate of c-Met protein expression in the residual cancer cells was higher in the experimental group,which indicated that hyperthermia could promote the expression of c-Met protein in residual cancer cells.Conclusion:1?The positive rate of c-Met protein expression was higher in the residual cancer cells HepG2 in vitro hyperthermia,and hyperthermia can promote the expression of c-Met protein in residual cancer cells.2? c-Met protein may be a new biomarker that suggests the recurrence,metastasis and efficacy of hepatocellular carcinoma.