Proteomic Study Of HCC And Evaluation Of Diagnostic Efficacy Of Several Differential Proteins Combinations

Objective In this study,we improve the mass spectrometry level of protein identification through improving protein separation technology and use the improved technology to explore the differentially expressed proteins in patients with HBV related hepatocellular carcinoma(HCC).After clinical verification of the differential proteins by commercial kit,evaluate the the efficacy of individual and joint detection in early diagnosis of hepatocellular carcinoma,and to explore new tumor markers for early diagnosis of hepatocellular carcinoma.Methods Collect the serum of 60 HBV-related HCC patients,70 cirrhosis patients associated to HBV and 70 patients with chronic hepatitis B who were all diagnosed originally in the liver medicine and surgery departments of Tianjin Third Central Hospital from January,2014 to February,2015 and the serum samples of 50 healthy human in preventive care for regular physical examination at the same time.Use the nano HPLC-MS technology to process,analysis and identify the serum samples.Then screen the differential expressed proteins though OMMSA software identifying and analyzing the relative expression quantity.Then run a clinical trial on the differential expressed proteins through commercialization kits.Compare HBV related HCC group and healthy group,HBV related HCC group and chronic hepatitis B group,HBV related HCC group and hepatitis B cirrhosis group and the different stages in HBV related HCC group to explore the value of these differential proteins in early diagnosis of liver cancer,and to find a new tumor marker for early diagnosis of hepatocellular cancer.Results(1)Use the improved proteomic technology,which used the Self-owned trypsin to hydrolyze the protein,isolate protein through one-dimensional capillary column and quantify accurately the proteins through LTQ Orbitrap XL mass spectrometer,at last this study screen out 3578 kinds of proteins through the OMSSA software.(2)Through statistical analysis select 10 differential proteins related to hepatocellular cancer,then run a clinical trial on the obvious four kinds(A1BG,AT-?,VTN and AFP).The outcomes of A1 BG and AT-? is consistent with the previous and have a higher diagnostic efficiency.But the mechanism of VTN shall to be discussed.(3)In health examination,A1 BG shall choose to be the hepatocellular carcinoma marker.For identification of hepatocellular carcinoma and hepatitis,AFP and AT-? have a higher diagnostic efficiency.For identification of hepatocellular carcinoma and liver cirrhosis,AFP and A1 BG shall be considered to combine to improve the diagnostic efficiency.Conclusions The application of self modified trypsin which can improve efficiency and two dimensional integrated chromatographic column technique which integrates the scx-strong cation exchange and C18 reversed phase liquid chromatography to eliminate the pollution of the samples and improve the repeatability shall improve the overall sensitivity of protein identification.Using the improved technology and reasonable statistical analysis method,we successfully screened out the differential proteins which can use in hepatocellular carcinoma early diagnosis and disease monitoring.Evaluate the diagnostic efficiency of differentially expressed protein,it can improve the dianostic rate of hepatocellular carcinoma if choose a right combination method,win valuable time of diagnosis and treatment for clinicians and at last improve the survival rate of patients with HCC.