Effects Of CXCL2 On The Biological Function Of SMMC-7721 Hepatocarcinoma

Background and objective:In China,Hepatocellular carcinoma is one of the most common malignancies,ranking secondly in the cause of cancer death across the country;the occurrence and development of cancer are closely related to its microenvironment of inflammation.With the development of molecular biology and bioinformatics,the same or different researchers have discovered that tumor cells can promote the formation of tumor inflammatory microenvironment in the process of co-evolution of microenvironment.Studying the relationship between high expression of pro-inflammatory cytokines and inflammation-cancer signaling pathways in the tumor microenvironment have became the focus of current researchs on tumor microenvironment.CXCL2 is a kind of chemokine,which can promote the chemotaxis of neutrophils,the growth and migration of tumor cells with the combination of specific receptor CXCR2.Our previous study found that differential secretory protein CXCL2 was found in the co-culture system of M2 macrophages and hepatocellular carcinoma cells,and the migration and invasion ability of hepatocellular carcinoma cells were reduced after neutralized the CXCL2 in the co-culture system,suggesting that CXCL2 had an impact on the biological function of hepatocellular carcinoma cells.At present,there are relatively few reports on the relationship between CXCL2 and the development of hepatocellular carcinoma.In this study we will further verify the effect of CXCL2 overexpression on the biological function of hepatocellular carcinoma cells,and provide the basis for the early diagnose of liver cancer,the new treatment plan and the improvement of patients ' prognosis.METHODS: Lentiviral vector over-expressing CXCL2 gene was used to infect SMMC-7721 hepatocellular carcinoma cells,that was set to lentivirus infection group;and the blank control group was the SMMC-7721 hepatocellular carcinoma cells without any infection treatment.The expression of CXCL2 gene mRNA and CXCL2 protein were detected by real-time fluorescence quantitative PCR(qRT-PCR)and Western Blot respectively.The cell viability was tested by CCK-8 to verify the cell proliferation ability.The cell invasion experiment(Transwell)and the cell scratch test were used to evaluate the metastasis and migration ability of hepatocellular carcinoma.The adhesion of cells was detected by cell adhesion experiment.The cell cycle was measured by flow cytometry technique.Statistical analysis was performed by SPSS23.0 statistical software.Results: The human SMMC-7721 hepatocellular carcinoma cells line stably overexpressing CXCL2 was successfully constructed.The expression of CXCL2 mRNA and CXCL2 protein in the lentivirus infected group were higher than that in the blank control group;the CCK-8 assay detected the cell proliferation ability and found that the proliferation of cells in the lentivirus infected group was relatively higher(P<0.05).The Transwell assay showed that compared with the blank control group,the number of cells passing through the stroma in the lentivirus infection group were relatively higher and the difference was statistically significant(P<0.05).The cell migration at 0h,8h,and 24 h after scratching was observed in the cell scratch test.The migration rate was been calculated.The results showed that the lentivirus infection group had higher cell migration rate from 8 h(8 h P= 0.01,24 h P = 0.035).Cell adhesion experiments revealed that the number of cell adhesion in the lentivirus infection group was lesser(P<0.05).Two groups of cell cycle test results showed that the proportion of G1 phase cells in the Lentiviral infection group was higher(P=0.034),the proportion of S phase was lower(P=0.017),and there was no significant change in the G2/M phase.Conclusion: The stable overexpression of CXCL2 gene in SMMC-7721 hepatoma cell can promote the proliferation of SMMC-7721 cells,enhance cell migration and invasion ability and inhibit the adhesion of cells through slow viral infection.According to the cell cycle results of the two groups,it is inferred that the cell cycle could be blocked during the G1 period.