The Study On The Detection Of Mir-26b Combined With GP73 In The Diagnosis Of Hepatocellular Carcinoma

Background and purpose:Hepatocellular carcinoma(hepatocellularcarcinoma,HCC)is one of the malignant tumorswith a high rate of malignancy,rapid progression and poor prognosis.Due to the lack of effective early diagnosis index,most liver cancer has been diagnosedinmiddle or advanced stage.Therefore,early diagnosis and early treatment is extremely important.In recent years,a lot of new serum markers of liver cancer have been found,and it is expected to be an ideal target for early diagnosis of liver cancer.Alpha fetoprotein(AFP)is an important index which most widelyused at present in clinical diagnosis of HCC,but in the diagnosis of HCC,AFP is still limited.Because in the clinical work about 40% of HCCpatients,AFP was at low level or negative.Therefore,to a certain extent,this will affect the clinical diagnosis of HCC.In our previous study,we found that the expression of mi R-26 b was significantly down regulated in HCC tissues,and correlated with the proliferation,invasion and metastasis of HCC.However,the expression of mir-26 b in peripheral blood and whether it can be used in the early detection of HCC need to be further validated.Golgi glycoprotein-73(GP73)Golgi,a type II transmembrane protein,mainly expressed in bile duct epithelial cells and little or no expression in liver cells.In acute and chronic hepatitis or hepatic fibrosis,the expression of GP73 is significantly increased,when in the development of liver Cirrhosis,the expression of GP73 is reach a higher level.Its expression reaches the peak when in HCC.This study focuses on the expression of mi R-26 b,GP73 and AFP in peripheral blood of people with normal,hepatitis and cirrhosis and HCC,and to analyze its sensitivity and specificity as the molecular diagnosis for HCC.The combined detection of AFP,mi R-26,GP73,in order to improve the diagnosis sensitivity and positive detection rate,may be a useful detection method for early diagnosis of HCC.Methods:1.The experiments were divided into 4 groups,including HCC group(n = 50),liver cirrhosis group(n = 40),chronic hepatitis group(n = 45)and healthy control group(n = 50).Real time fluorescent quantitative polymerase chain reaction(q RT-PCR)was used to detect the expression level of mi R-26 b in whole blood of all groups.2.Enzyme linked immunosorbent assay(ELISA)was used to determine the concentration of GP73 in serum.And the level of alpha fetoprotein(AFP)was measured by chemiluminescencemicroparticle immunoassay.3.The optimal cutoff values of mi R-26,GP73 and AFP were determined using the receiver operating characteristic curve(ROC curve)in peripheral blood.Early diagnostic values for HCC of this three markers and combined detection(mi R-26-GP73?mi R-26-AFP?GP73-AFP?mi R-26-GP73-AFP)were analyzed by ROC curve.Results:1.Compared with the liver cirrhosis group,chronic hepatitis group and healthy control group,mi R-26 b showed low level expression in peripheral blood of HCC,the difference was statistically significant(P<0.05).There was no significant difference between liver cirrhosis group,chronic hepatitis group and healthy control group(p> 0.05).2.The serum GP73 level in the HCC group(234.58(166.50-309.49)ng/ml)was significantly higher than that in liver cirrhosis group(133.25(46.50-146.60)ng/ml),chronic hepatitis group(141.01(82.77-149.56)ng/ml)and control group(43.50(37.06-49.63)ng/ml),and the difference was statistically significant(P<0.05).There was no significant difference between the liver cirrhosis group and the chronic hepatitis group(p> 0.05).3.The ROC curve of mi R-26 bshowed that AUCwas 0.936,the optimal cut-off value(cutoff value)was 0.888,the diagnostic sensitivity and specificity of HCC were 92% and 87.5%;GP73 ROC curve showed that AUC was 0.913,cutoff value was 144.82ng/ml,and the sensitivity and specificity were 84% and 77%;respectively,AFP HCC in the diagnosis of AUC was 0.718,the optimal cut-off value of 27.99ng/ml,sensitivity and specificity were 62% and 68.9%.Among them,the highest sensitivity and specificity were mi R-26 b.Also,the largest AUC wasmi R-26 b.4.Among 28 cases with low serum AFP concentration(AFP < 400 ng/ml)in HCC,24 patients with mi R-26 b positive(cutoff = 0.888)account for 85.7%(24/28)of AFP patients with low concentration of HCC;There were 16 patients with GP73 positive(GP73 ?150 ng / ml),accounting for 57.1%(16/28)of AFP low level of HCC.The positive detection rate of mi R-26 b and GP73 was much higher than that of AFP(P<0.01).5,With GP73?150ng / ml,AFP?400ng / ml as the diagnostic criteria,the sensitivity of mi R-26 b and GP73 in HCC was 86.0% and 78.0%,respectively,which was significantly better than that of AFP(44.0%)(P <0.01),but their specificitiesare worse than AFP.From the ROC curve to observe the diagnosis of HCC,mi R-26 b,GP73 and AFP curve area was significantly different(P<0.05).5.Mi R-26 b and GP73 are superior to AFP in the diagnosis of HCC,and to a certain extent they can make up for the missed diagnosis rate of AFP,and may become a new serological marker of liver cancer.Combined detection of mi R-26 b,GP73,AFP can improve the accuracy of early diagnosis of liver cancer.Conclusion:1.The expression of mi R-26 b was decreased in peripheral blood of HCC group,which may serve as a new molecular marker for the diagnosis of HCC.When combined with serum AFP detection,mi R-26 b has important clinical value for the diagnosis of HCC.2.The expression of GP73 in the serum of HCC group was significantly increased,suggesting that it can be used as a kind of significant HCC serum markers,and provide a marker for tumor diagnosis.3.The combined detection of mi R-26 b,GP73,AFP in blood has positive significance for the early diagnosis of HCC,and it can also be used for screening of high-risk population of liver cancer.