Clinical Application Of Chitinase 3 Like Protein 1 (CHI3L1) Detection In Hepatocellular Carcinoma And Liver Cirrhosis

Primary liver cancer(PLC)is one of the most common malignant tumors.It mainly includes three types: hepatocellular carcinoma(HCC),intrahepatic cholangiocarcinoma(ICC)and HCC-ICC mixed cell carcinoma.Hepatocellular carcinoma accounts for 85-90 percent.According to the report of cancer epidemic situation in China in 2015,the incidence of liver cancer was as high as 26.92/100000,ranking the fourth among malignant tumors in China.The mortality rate was 23.72/100000,ranking second in China,only next to lung cancer.The main risk factors of primary liver cancer are hepatitis virus infection,alcohol,aflatoxin in food intake and so on.Detection of serum markers is usefull mean for early detection and diagnosis of liver cancer.In recent years,many protein markers related to the occurrence and development of tumor have been found in basic research,and some of them have entered clinical application.However,the application value of these new markers needs further clinical research.,As a kind of secreted glycoprotein,Chitinase 3 like protein 1(CHI3L1)expression increases in many solid tumors and inflammatory diseases.In this study,we dectected the expression of CHI3L1 in tissues and peripheral blood of HCC patients to explore the clinical application of CHI3L1 in primary liver cancer.Part one: expression of CHI3L1 in hepatocellular carcinoma tissuesFirstly,we analyzed the expression and prognosis value of CHI3L1 in tumor tissues and matched adjacent tissues of 50 HCC patients from TCGA database.We found that the expression of CHI3L1 in tumor tissues were higher than that in adjacent tissues(t = 2.691,P = 0.010).High expression of CHI3L1 in tumor tissues was positively correlated with poor prognosis of HCC patients.Then,we decided to investigate the expression of CHI3L1 in HCC.In order to providing proof for the clinical application of CHI3L1 expression in HCC patients,Western blot(WB),reverse transcription polymerase chain reaction(RT-PCR)and tissue microarray immunohistochemical staining were used to dectect the expression of CHI3L1 in tumor tissues and paired adjacent tissues of HCC patients.There was no significant difference in CHI3L1 transcription level(RT-PCR)and protein level(WB)between cancer tissues and adjacent tissues of 20 patients with HCC(P = 0.199,0.064).The CHI3L1 staining index of adjacent tissues was higher than that of tumor tissues(z =-2.343,P = 0.019).There was no significant difference in the expression of CHI3L1 between HCC patients with cirrhosis and those without cirrhosis(z =-1.781,-1.030,P = 0.075,0.303),either in tumor tissues or adjacent tissues.We analyzed relationship between pathological features and CHI3L1 staining index,found that chi3l1 staining index was related to tumor size,daughter stove(satellite stove)and microvascular invasion(z = 2.387,2.112,2.092,P = 0.017,0.035,0.036).The survival analysis showed that there was a significant correlation between satellite stove,tumor stage,tumor size,AFP level,white blood cell count,and plasma fibrinogen level(P = 0.012,0.028,0.001,0.039,0.034,0.041,0.037).In addition,difference between CHI3L1 staining index of adjacent tissues and cancer tissues showed significant value in survival in HCC patients,The patients with high △ CHI3L1(paracancerous tissue-cancer tissue)has poorer survival(P = 0.011).Part two: Changes and clinical significance of CHI3L1 in peripheral blood of patients with liver cirrhosis and hepatocellular carcinomaWe detected CHI3L1 protein in peripheral blood of patients with liver cirrhosis and hepatocellular carcinoma to analyzed its clinical management values.The level of CHI3L1 in HCC、LC、HC groups is 118.2(74.9,201.0),195.8(103.3,330.4)、46.8(30.7,66.4)μg/L,respectively,significant differences between the three groups were observed.In LC group,CHI3L1 protein was positively correlated with FIB-4(r = 0.237,P = 0.013).The level of CHI3L1 in patients with Child Pugh B and C was higher than that of Child Pugh A,which may showe that level of CHI3L1 relating to liver fibrosis and liver function in LC patients;ROC curve was used to analyze diagnostic value of CHI3L1 in LC patients.the area under the curve(AUC)was 0.934,which indicated that CHI3L1 has diagnostic efficiency for LC.Patients in HCC group were divided into two groups: HCC with cirrhosis and HCC without cirrhosis,no significant difference was observed between the two groups(P =0.775),which was consistent with the results of tissue microarray in the first part.The correlation between serum CHI3L1 level and clinical laboratory indexes was analyzed.Serum CHI3L1 level was positively correlated with hyaluronic acid(HA),type Ⅲprocollagen N-terminal peptide(PCⅢNP),type Ⅳ collagen(Ⅳ-C),FIB-4(r = 0.202,0.159,r = 0.299,0.221,P < 0.05 respectively),and negatively correlated with ALB(r =-0.326,P < 0.001),positively correlated with PT(r = 0.160,P = 0.010)and AST(r = 0.138,P = 0.009).The correlation between serum CHI3L1 and pathological features of HCC patients was analyzed.The serum CHI3L1 was positively correlated with the maximum tumor diameter(r = 0.284,P < 0.05)Although there was no correlation between serum CHI3L1 level and MVI in HCC patients,we found that the level of CHI3L1 in MVIpositive group was significantly higher than that in MVI negative group,in AFP negative HCC patients.Serum levels of CHI3L1 of 30 patients with HCC after surgical resection were significantly higher than basic line,which may be related to tissue repair and liver regeneration.The level of CHI3L1 in peripheral blood of LC patients was higher than that of HCC patients.Serum CHI3L1 level in HCC patients was related to liver fibrosis markers,liver function indexes,tumor size and tumor stage.Detection of serum CHI3L1 level is helpful to clinical hierarchical management of HCC patients.To summarize,we found that both CHI3L1 m RNA and protein was highly expressed in hepatocellular carcinoma tissues,and the level of CHI3L1 in peripheral blood of patients with liver cirrhosis and hepatocellular carcinoma was also increased.Compared with healthy controls,peripheral blood CHI3L1 has good diagnostic efficiency for liver cirrhosis.Detecting CHI3L1 is helpful for disease stratification and clinical management of HCC patients.Fibrosis and precancerous changes exist in the adjacent tissues of HCC patients,which may be the influencing factors of inconsistent reports on HCC and LC in the current literature.The influence of tumor,inflammation,fibrosis and other factors should be considered in the clinical application of CHI3L1.