The Effect Of Recombinant High Mobility Group Box 1 On Invasion And Metastasis Of Esophageal Cancer Cells

Esophageal cancer is a common malignant tumor of digestive system.It includes squamous cell carcinoma and adenocarcinoma.Objection: The order is to investigate the effect of recombinant High mobility group box 1 on migration and invasion of human esophageal cancer TE13 cell line and explore its possible mechanisms.Method: The experiment is divided into 4 groups,different doses(0,100,200,400ng/ml)of recombinant HMGB1 were used to stimulate esophageal cancer TE13 cells for 24 h.Group 0ng/ml was the control group,group 100ng/ml,200ng/ml and 400ng/ml were the experimental group.The proliferation was detected by CCK8.Scratch test was used to evaluate the effect of invasion and metastasis.P65 and MMP2 expression was detected by fluorescence quantitative PCR and Western-blot method.Results:1.CCK8 results show that: the OD value was 3.02±0.11,2.93±0.11,2.98±0.14,2.98±0.07 respectively.The OD values of the three treatment groups(100ng/ml,200ng/ml,400ng/ml)were not statistically different from those of the control group(0ng/ml).Also the difference was no significance between any two treatment groups(P>0.05).2.Cell scratch test showed that: after different concentrations of HMGB1(100ng/ml,200ng/ml,400ng/ml)stimulated esophageal cancer cell for 24 h,compared with the control group,the cell migration distance increased(P<0.05).The cell migration distance in group 200ng/ml and group 400ng/ml was higher than that in group 100ng/ml(P<0.05).There was no significant difference between group 200ng/ml and group 400ng/ml(P>0.05).3.Fluorescence quantitative PCR test results showed that:1)The relative content of P65 mRNA expression was 1.00±0.04,1.53±0.05,2.19±0.03,2.12±0.03.The relative content of p65 mRNA in the treatment group was higher than that in the control group(0ng/ml).There was no significant difference between group 200ng/ml and group 400ng/ml(P>0.05),but in the else treatment groups the difference was statistical significant between any two groups(P<0.05).2)The relative content of MMP2 mRNA expression was 1.00±0.09,1.11±0.02,2.03±0.07,1.96±0.06.In group 200ng/ml and group 400ng/ml,the expressions of MMP2 mRNA were higher than that in control group(0ng/ml)and group 100ng/ml,the difference was statistically significant(P<0.05),and there was no statistical difference between the other groups(P>0.05).4.Western-blot test results showed that:1)The relative content of total P65 protein expression was 0.35±0.02,0.79±0.03,0.92±0.05,0.87±0.05.The total P65 protein expression in the treatment group was higher than that in the control group(P<0.05).2)The relative content of total MMP2 protein expression was0.22±0.03,0.32±0.08,0.58±0.14,0.55±0.14.In group 200ng/ml and group 400ng/ml,the expression level of MMP-2 protein was significantly higher than that in control group(P<0.05),but there was no significant difference between group 200ng/ml and group 400ng/ml(P>0.05),the difference between the other treatment groups was statistically significance(P<0.05).The results showed that HMGB1 stimulation could upregulate NF-?B P65 and MMP2 protein expression.Conclusion: HMGB1 can promote the invasion and metastasis of esophageal cancer TE13 cells and regulate the expression of P65 and MMP2 at RNA and protein levels.HMGB1 may activate the P65 signaling pathway and then activate MMP2,thus promoting metastasis and invasion of esophageal cancer cells.