ABSTRACT
BACKGROUND/AIMS: We conducted a case-control study in China to clarify the association between XRCC1-Arg-399Gin polymorphism and HCC risk. METHODOLOGY: A total of 150 cases and 158 controls were selected from May 2008 to May 2010. XRCC1-Arg399Gin and XRCC3-Thr241Met polymorphisms were based upon duplex polymerase-chain-reaction with the confronting-two-pair primer (PCR-CTPP) method. All analysis was performed by using the STATA statistical package. RESULTS: A significant increased risk of HCC was associated with XRCC1 399Arg/Gin, and a heavy risk of HCC was also found in individuals with XRCC3 241Met/Met genotypes. A significant association was found between positive HBsAg and Arg/Gin. XRCC3 Thr/Met genotypes had a significant positive association with HBsAg (+) and a heavy risk of HCC was found in HBsAg (+) individuals with XRCC3 Met/Met genotype. Individuals carrying XRCC1 Gin/Gin genotypes showed significantly lower median survival than XRCC1 Arg/ Arg genotypes and significant hazard ratio (HR=l.38, 95% CI=l.04-1.84) was found. Meanwhile, we found a moderate HR for XRCC3 Thr/Met (HR=l.96, 95% CI=l.23-3.15) and a heavy HR for XRCC3 Met/Met (HR=2.98, 95% CI=1.77-7.54). CONCLUSIONS: In conclusion, we observed that XRCC1-Arg399Gln and XRCC3-Thr241Met polymorphism is associated with susceptibility to HCC and XRCC1 Gin allele and XRCC3 Met allele genotype showed significant poor prognosis of HCC.
Subject(s)
Carcinoma, Hepatocellular/genetics , DNA Repair , DNA-Binding Proteins/genetics , Liver Neoplasms/genetics , Polymorphism, Genetic , Adult , Alcohol Drinking/adverse effects , Biomarkers/blood , Carcinoma, Hepatocellular/etiology , Carcinoma, Hepatocellular/mortality , Carcinoma, Hepatocellular/pathology , Carcinoma, Hepatocellular/therapy , Case-Control Studies , Chi-Square Distribution , China , Female , Genetic Predisposition to Disease , Hepatitis B/complications , Hepatitis B/diagnosis , Hepatitis B Surface Antigens/blood , Humans , Kaplan-Meier Estimate , Liver Neoplasms/etiology , Liver Neoplasms/mortality , Liver Neoplasms/pathology , Liver Neoplasms/therapy , Logistic Models , Male , Middle Aged , Odds Ratio , Polymerase Chain Reaction/methods , Prognosis , Proportional Hazards Models , Risk Factors , X-ray Repair Cross Complementing Protein 1ABSTRACT
Objective To prepare the PSMA7 hybridoma cell lines using classical hybridoma technique and to perform purification and elementary identification of the monoclonal antibodies(McAbs)against PSMA7 antigen for further study.Methods BALB/c mice were immunized with purified recombinant PSMA7 protein.Once the antibody titer of blood taken from mouse tail reached 1∶6 400,a cell fusion was performed between mouse splenic cells and myeloma cells(Sp2/0),and then the hybridoma cell lines secreting monoclonal antibodies against PSMA7 antigen were screened by indirect enzyme linked immunosorbent assay(ELISA).The immunoglobulin subtypes and titer of the monoclonal antibodies against PSMA7 antigen were identified and measured by Western blot analysis and enzyme linked immunosorbent assay,respectively.Results After cell fusion and subcloning,two hybridoma cell lines that stably secreted monoclonal antibodies against PSMA7 antigen were successfully obtained:B013 and B001,which belong to the subtypes of IgG1.The antibody titers in the hybridoma culture supernatant were 1∶128 and 1∶256,and those in the ascites fluid were 1∶25 600 and 1∶32 000,respectively.Western blot analysis and immunohistochemistry showed that the two antibodies can specifically bind with PSMA7 antigen derived from human eucaryotic cells or tissue.Conclusions Two monoclonal antibodies against PSMA7 antigen with high titers and specificity have been successfully prepared.These antibodies can be used for the identification of PSMA7 protein,and may be a useful tool for studying the biological properties of PSMA7 protein.
