The Molecular Mechanism Of CXCL8 And Its Receptors CXCR1 And CXCR2 In Transformation From Inflammation To Carcinoma In The Patients With Hepatocellular Carcinoma

Objective To study the expression level of CXCL8 and its receptor CXCR1/2 in peripheral blood mononuclear cells(PBMCs)and polymorphonuclearces(PMNs)of patients with hepatocellular carcinoma(HCC),to further detect the content of CXCL8 and its receptor CXCR1/2,to analyze the relationship between serum enzymology and viral load,and to explore the role of CXCL8 and its receptor CXCR1/2 in the molecular mechanism of HCC-cancer transformation.Methods Based on the criteria for diagnosis and treatment of primary HCC issued in 2017 in China,42 patients with HBV-related HCC were selected.According to the size of nodules examined by imaging,the patients were divided into two groups:the massive group(d>10cm)and the nodule group(d<5cm).Serum CXCL8 protein level was determined by ELISA;PBMCs and PMNs were separated from peripheral blood by density gradient separation,and cRNA was amplified after retroviral transcription by Triozl extraction.CXCL8,CXCR1 and CXCR2 in PBMCs and PMNs of patients with HCC were detected by qPCR.GAPDH was taken as internal reference and the final content was represented by 1gDNA/1gGAPDH.IBM SPSS Statistics 20.0 was used to analyze their interaction with viral load,serum GGT and ALT.Relationship.The expression of CXCL8 and its receptor CXCR1/2 and their relationship were observed by strept avidin-biotin complex(SABC)immunohistochemistry.Results The levels of serum enzyme markers,CXCL8 protein and CXCL8-CXCR1/2 from PBMCs,PMNs in peripheral blood were significantly increased in HCC(P<0.001).There was significant difference in ALT expression between massive and nodular HCC(P=0.0013).The level of serum CXCL8 in massive group was significant correlation with ALT and GGT(r=0.763;r=0.741),serum CXCL8 was highly correlation with ALT and GGT in nodular HCC(r=0.832;r=0.887),serum CXCL8 were low correlation(r=0.478)and significant correlation(r=0.642)with GGT/ALT in patients with massive and nodular HCC.The mRNA levels of CXCL8,CXCR1 and CXCR2 in peripheral blood of patients were shown as follows,such as(massive-PBMCs:0.80�0.30,massive-PMNs:0.93�0.33;nodular-PBMCs:0.89�0.28,nodular-PMNs:1.04�0.29),(massive-PBMCs:0.72�0.28,massive-PMNs:0.90�0.26;nodular-PBMCs:0.88�0.18 nodular-PMNs:1.05�0.17),(massive-PBMCs:0.92�0.20,massive-PMNs:0.73�0.21;nodular-PBMCs:1.08�0.20 nodular-PMNs:0.92�0.20).There were significant differences in CXCL8 and CXCR1 mRNA between the control group and HCC group(P<0.001),but there was no significant difference in CXCR2 mRNA between the two groups(PBMCs:P=0.160;PMNs:P=0.400).There was no significant difference in HBV-DNA in PBMCs and PMNs of massive and nodular hepatocellular carcinoma(PBMCs:P=0.055;PMNs:P=0.710).The expression of CXCL8-CXCR1 mRNA in PBMCs and PMNs of massive and nodular HCC were highly correlated(massive-PBMCs:r=0.930,massive-PMNs:r=0.912;nodular-PBMCs:r=0.806,nodular-PMNs:r=0.802),and CXCL8-CXCL8-CXCR2 mRNA were low correlated(massive-PBMCs:r=0.369,massive-PMNs:r=0.414;nodular-PBMCs:r=0.579,nodular-PMNs:r=0.414),there were no statistical significance(P>0.05).HBV-DNA in PBMCs and PMNs were positively correlated with CXCL8 and CXCR1,but no significantly(PBMCs-CXCL8:r=0.269,PBMCs-CXCRl:r=0.057;PMNs-CXCL8:r=0.517,PMNs-CXCR1:r=0.335).The levels of CXCL8,CXCR1 and CXCR2 in PMNs were higher than those in PBMCs,the levels of intracellular CXCL8,CXCR1 and CXCR2 in patients with HBV-DNA(+)were higher than those in patients with HBV-DNA(-).CXCL8 and CXCR1 in PBMCs and PMNs were highly correlated with GGT/ALT(PBMCS-CXCL8:r=0.900,PBMCs-CXCR1:r=0.837;PMNs-CXCL8:r=0.865,PMNs-CXCR1:r=0.849);CXCL8,CXCR1 in nodular HCC PBMCs and PMNs were significantly correlated with GGT/ALT(PBMCs-CXCL8:r=0.704,PBMCs-CXCXR1:r=0.574,PMNs-CXCL8:r=0.694,PMNs-CXCR1:r=0.673);CXCR2 in PBMCs and PMNs of massive HCC showed weak correlation with GGT/ALT(r=0.281)and low correlation(r=0.357),CXCR2 in PBMCs and PMNs of nodular HCC were significantly correlated with GGT/ALT(r=0.566)and low correlated with GGT/ALT(r=0.426).SABC immunohistochemical staining showed that CXCL8 and CXCR1 were highly expressed in liver cancer tissues of HCC(P<0.01),and poor expression of CXCR2 in those of liver cancer tissues.CXCL8 and its receptors CXCR1/2 were diffusely expressed in the cytoplasm of hepatoma cells,inflammatory cells and vascular endothelial cells.Conclusion The expression of CXCL8 and its receptor CXCR1 in peripheral blood and local cancer tissues of HCC increased,which is positively correlated with concentration of ALT,GGT,GGT/ALT and HBV-DNA load.The expression of CXCL8-CXCR1 axis in PMNs is higher than that in PBMCs.The high levels of CXCL8 and its receptor CXCR1 can be increased in local lesions,which play an important role in the infiltration and growth of HCC,malignant hyperplasia and transformation from inflammation to cancer.However,the effect of CXCR2 may be relatively weaker.Figure[26]table[22]reference[54]...