| BackgroundPrimary liver cancer(PLC)is one of the most common malignant tumors leading to human death around the world.Data from the World Health Organization’s International Agency for Research on Cancer(IARC)show that in 2020,905,677 new cases of liver cancer were reported worldwide,accounting for 4.7%of the total number of new cancers,ranking sixth,and 830,180 cases of liver cancer deaths,accounting for 8.3%of the total number of cancer deaths,ranking third.Hepatocellular carcinoma(HCC)is the most common pathological type,accounting for 85%to 90%.There are many pathogenic factors of liver cancer.At present,it is believed that the common pathogenic factors of liver cancer mainly include hepatitis virus infection,cirrhosis,consumption of aflatoxin-contaminated food,excessive drinking and so on.Currently,many studies have proved that hepatitis B virus(HBV)is the main cause of liver cancer.Although the vaccination of hepatitis B vaccine has led to a gradual decline in the number of new cases of hepatitis B in China,an increasing number of patients infected with HBV have developed into Chronic hepatitis B(CHB),which brings a heavy burden to the health of the Chinese people.Chronic HBV carriers have a 10 to 25-times greater lifetime risk of developing HCC compared with uninfected people.Therefore,the study of hepatitis B virus infection-associated liver cancer is a hot spot and difficult point.Currently,studies have confirmed that hepatitis B virus mediates the occurrence of HCC through a variety of pathways,among which,the epigenetic mechanism has received more and more attention,and DNA methylation is the main mechanism of epigenetic control.Studies have shown that global hypomethylation and selective hypermethylation of DNA can occur in the early stage of tumor,which silencing the promoters of some tumor suppressor genes and reducing gene expression,thus promoting the occurrence and progression of tumors.Currently,early screening methods for HCC are mainly carried out by tumor indicators such as liver Ultrasound(US)and serum alpha-fetoprotein(AFP).If liver ultrasound or AFP and other tumor indicators are found to be abnormal,we need to conduct further examination to confirm the diagnosis,including:Computer tomography(CT),Magnetic resonance imaging(MRI),digital subtraction angiography(DSA),positron emission computed tomography(PET-CT),biopsy and other auxiliary examinations.Among them,AFP,as the most commonly used serological index in the clinical diagnosis of liver cancer,has some limitations.A number of studies have confirmed that the level of AFP is also increased in many other diseases.Therefore,the serological markers of liver cancer still need further research and exploration.Interleukin-8 is a member of the chemokine family,which was first discovered by Kownatzki et al.A number of studies have shown that IL-8 not only plays a role in chemotactic neutrophil-mediated inflammation,but also plays a role in chemotactic neutrophil-mediated inflammation.It can also promote the development and metastasis of tumors by increasing the activity of NF-κB,increasing the angiogenesis in tumors,recruiting immunosuppressive cells and changing the immune microenvironment.Previous studies have shown that IL-8 is highly expressed in liver cancer patients,and there are many studies on the mechanism of its high expression,but there is no study on the role of IL-8DNA methylation in liver cancer at presentObjectiveBy detecting the methylation level of IL-8 promoter in peripheral blood mononuclear cells(PBMC)of HCC patients,CHB patients and Healthy controls(HCs),we can explore its diagnostic value in HCCMethodA total of 64 patients with HCC,34 patients with CHB,and 26 patients with HCs were enrolled.Fastering peripheral blood of the research subjects was collected to extract mononuclear cells from the peripheral blood.DNA was extracted from the mononuclear cells by Trizol method,and the extracted DNA was methylated and modified.Methylight technology was used to detect the methylation level of IL-8 DNA in the samples.Percentage of methylated reference(PMR)of IL-8 was calculated,and the relationship between PMR and other clinical indicators was explored by statistical methods.Results1.Contrast of the basic clinical characteristics of the three groups:We found that there were no significant differences in age and gender among the subjects in the HCC group,CHB group and HCs group(P>0.05).There were no significant differences in HBeAg,HBV DNA,ALT,AST,GGT,ALP,TBil,ALB,PTA and PLT between HCC group and CHB group(P>0.05).AFP value in the HCC group was higher than that in the CHB group,which was statistically significant(P<0.05).2.Contrast of the methylation degree of IL-8 promoter among the three groups:The methylation degree of IL-8 promoter in HCC group,namely PMR(0.680±0.62%),was significantly lower than that in CHB group(2.15±0.89%,P<0.05)and HCs group(21.40±4.81%,P<0.05).The methylation level of IL-8 promoter in CHB group was also significantly lower than that in HC group(P<0.05).3.Correlation between IL-8 promoter methylation degree and clinical data in the HCC group:In patients with HCC,The methylation degree of IL-8 promoter was not significantly correlated with ALT(P=0.673),AST(P=0.792),GGT(P=0.508),ALP(P=0.915),ALB(P=0.517),PTA(P=0.090),PLT(P=0.232)and AFP(P=0.983)Correlation was positively correlated with TBIL degree(r=0.379,P=0.002).4.Correlation between IL-8 promoter methylation and clinicopathological features in the HCC group:In patients with HCC,The methylation degree of IL-8 promoter was correlated with gender(P=0.855),age(P=0.660),smoking(P=0.973),alcohol consumption(P=0.957),HBeAg(P=0.766),tumor size(P=0.314),tumor number(P=0.110),vascular invasion(P=0.151),and there wereThere was no significant correlation between distant metastasis(P=0.315)and pathological stage(P=0.847).5.The level of serum IL-8 promoter methylation and the diagnostic value of AFP in HCC:AFP had a sensitivity of 42.2%,a specificity of 90.9%,and a cutoff value of 150.00ng/mL for distinguishing HCC from CHB.The IL-8 promoter methylation index PMR had a sensitivity of 82.4%,a specificity of 90.6%,and a cutoff value of 1.45%for distinguishing HCC from CHB.The AUC of IL-8 promoter methylation index PMR for distinguishing HCC and CHB was 0.916(Se 0.030,95%CI 0.857-0.975),which was significantly higher than that of AFP(Se 0.055,95%CI 0.563-0.777),and the difference was statistically significant(P<0.05).Conclusions1.The methylation degree of IL-8 promoter in HCC patients was significantly lower than that in CHB group and HCs group.It suggests that the hypomethylation of IL-8 promoter may be one of the reasons for the high expression of IL-8 gene in HCC patients.2.In HCC patients,the methylation level of IL-8 promoter was positively correlated with total bilirubin(TBIL),suggesting that IL-8 gene expression may be affected by bilirubin.The methylation degree of IL-8 promoter in HCC patients was not significantly correlated with ALT,AST,AFP and other clinical parameters,suggesting that the methylation degree of IL-8 gene was less affected by other factors and was relatively stable.3.In HCC patients,the degree of IL-8 promoter methylation was not significantly correlated with the clinicopathological characteristics such as tumor size,number,vascular invasion or distant metastasis,and pathological stage,suggesting that the degree of IL-8 promoter methylation could not reflect the severity of the lesion.4.The diagnostic accuracy of IL-8 promoter methylation in HCC patients is higher than that of AFP,suggesting that the PMR value of IL-8 promoter methylation can be used as a non-invasive marker for the diagnosis of HCC in patients with chronic hepatitis B,providing a new direction for the diagnosis of HCC. |
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