Expression And Clinical Significance Of Caspase-3,Bim And C-Cbl In Gastric Carcinoma

IntroductionGastric cancer is a common gastrointestinal carcinoma in China,whose genesis is a poly stages and poly factors development process.The disequilibrium between cell proliferation and apoptosis is the important factor in inducing carcinoma absoluteness growth.For the past few years,people pay more and more attention to carcinoma apoptosis.Apoptosis named as programmed cell death(PCD).Carcinoma genesis is associated with overexpression of oncogene in controlling cell proliferation and with hypsoexpression of Bcl-2 gene as mutation and unactivation of antioncogene induced apoptosis.The important hypothesis of cell canceration is unduly termination of apoptotic mechanism.The restrain of apoptosis is significant effect to carcinoma genesis.The three targets,Caspase-3,Bim and c-Cbl,are closely correlated with gastric carcinoma genesis and prognosis.Caspase-3,which the important member of Caspase family and the most key effector molecule in apoptotic mechanism,participates physiological and pathological death procedure and is payed closed attention in apoptpsis research,but the relationship researchs of Caspase-3 and carcinoma biological behaviour are few.Bim,which only-BH3 domain protein in Bcl-2 family,processes activitiest promoting apoptosis and tumor-inhibiting factor and its physiological functions are correlated with its functions of regulating apoptosis. c-Cbl,which the important member of Cbl family,may mediate degradation of acceptor and nonreceptor tyrosine kinase and down-regulate protein activity as ubiquitin ligase E3 which is correlated with apoptosis.There was no report on combinated determination of Caspase-3,Bim and c-Cbl both in domestic and foreign countries.So this study was designed to investigate the expression of Caspase-3,Bim and c-Cbl and their clinical significance in gastric carcinomas.Methods1,Patients124 gastric cancer cases underwent completely resected operation,which were enrolled from the first affiliated hospital of China Medical University(CMU)and Liaoning Cancer Hospital from October 2005 to August 2006,and none of them underwent chemotherapeutics before operation.All the cases included 92 male and 32 female patients with median age of 60 years,ranged from 24 to 85 years.The histologic type of all cases is adencarcinoma with 11 cases of well differentiation(8.9%),29 moderated diferentiation(23.4%)and 84 poor diferentiation (67.7%).There were 76 cases with muscular layer invasion(61.3%)and 48 placenta percreta invasion(38.7%).There were 100 cases with lymph node metastasis (80.6%).There were 17 cases in stageⅠ(13.70;),46 in stageⅡ(37.1%),37 in stageⅢ(29.8%)and 24 in stageⅣ(19.40;).24 normal tissue cases were selected as controls, which were 5 centimeters far from gastric cancer tissues.2,MaterialsThe polyclonal antibodies against Caspase-3 was purchased from Santa Cruz corporation and the monoclonal antibodies against c-Cbl was purchased from BD corporation.Bim,which the polyclonal antibodies,S-P immunohistochemical kits and DAB substrate solution were purchased from Fuzhou Maixin Biotechnology,Inc.3,MethodsImmunohistochemical staining S-P method was performed on tissue microarray sections made of 124 cases of gastric carcinoma and 24 cases of paracancerous tissue. All slides were treated with citric acid buffer(PH=6.0)to recover the antigen activity in autoclave.4,AssessmentOutcome definition of Caspase-3 and Bim and were stained yellow particles in cytosol,outcome definition of c-Cbl was stained yellow particles in membrane and cytosol.Judging reference is followed by dyeing cells numbers and intensity,which product 0～1 as negative,2～4 as weakly positive,≥5 as strongly positive.5,Statistical analysisStatistical methods Chi-square test,Spearman rank correlation test and Fisher exact test were performed by SPSS software version 13.0.The statistical difference was considered significant when P＜0.05.Results1,The expression of Caspase-3,Bim and c-Cbl in patients with gastric carcinomaThe positive expression rates of Caspase-3 and Bim in gastric carcinoma were 45.2%and 37.9%,while the positive expression rate of c-Cbl in gastric carcinoma was 71.0%.2,The relationship between expression of Caspase-3,Bim and c-Cbl and clinicopathologic parameters in patients with gastric carcinomaThe positive expression rates of Caspase-3 was correlated with differentiation (P＜0.05);the positive expression rate of Bim and c-Cbl were correlated with TNM staging and degree of invasion(P＜0.05).3,The relationship between Caspase-3,Bim and c-CblThere was a positive relationship between the expression of Caspase-3 and Bim (r_s=0.403,P＜0.05),there was a negative relationship between the expression of Caspase-3 and c-Cbl as well as Bim and c-Cbl(r_s=-0.185,r_s=-0.442,P＜0.05)Conclusion1,The positive expression rates of Caspase-3 and Bim in gastric carcinoma were low,respectively,while the positive expression rate of c-Cbl in gastric carcinoma was high.2,The positive expression rates of Caspase-3 was correlated with differentiation,the positive expression rate of Bim and c-Cbl were correlated with TNM staging and degree of invasion.3,There was a positive relationship between the expression of Caspase-3 and Bim,,while there was a negative relationship between the expression of Caspase-3 and c-Cbl as well as Bim and c-Cbl.None of them was associate with age,sex,lymph node metastasis.