The Screening And Identification Of Associated Protein With Gallbladder Cancer

Background:Primary carcinoma of the gallbladder (PCG) is a highly malignant tumor. And the morbidity seem step up year by year.Historically, the 5-year survival of advanced stage Gallbladder cancer are reported to be<5%. However, in the case of early GBC in which the depth of cancer invasion is limited to the lamina propria or the muscle layer of the gallbladder wall without nodal involvement, hematogenous metastasis or peritoneal implantation, a 5-year survival rate of >80% can be expected even when a simple cholecystectomy is performed. But, those gallbladder cancer in the early stage which lack specificity marker have no special clinical manifestation almost found accidental. Cholecystolithiasis affiliate chronic cholecystitis (CC) is one of its major risk factors.Sing stone diameter>3cm verify one of imprtant cause with PCG.Gallbladder cancer which affilinated Cholecystolithiasis are high to 94%. Objective:To identify the proteins involved in PCG carcinogenesis, wishing find special maker associated with gallbladder cancer dignosis,treatment and prognosis,So as to more comprehend pathogenesis of PCG Methods:we firstly employed two-dimensional electrophoresis (2-DE) coupled with MAILDE-TOF MS/MS analysis to identify differentially expressed proteins among PCG ,adjacent nontumor tissue samples, cholecystolithiasis Samples ,and nomal gallbladder samples.from 10 gallbladder cancer patients were analyzed. A total of 21 spots were significantly up-regulated (ratio≥2, p≤0.01) in tumor samples, while 45 spots were down-regulated (ratio≤-2, p≤0.01). Twenty-one gene products were identified among these spots.with 6 of these being reported for the first time in PCG, Of the 6 newly implicated proteins.Altered expression of representative proteins including Moesin, AnnexinA4, Retinal dehydrogenase1, Vimentin, Heat shock protein 90βwas validated by RT-PCR and Western blotting. Moesin, Annexin A4 was demonstrated to be up-regulated in cancer at both the mRNA and protein levels, adversely Retinal dehydrogenase 1showed in cancer. Therefore, Moesin, Annexin A4, Retinal dehydrogenase 1 were chosen for detailed analysis. Results: There are 66 spots associate gallballder cancer contrast with adjacent nontumor tissue samples and cholecystolithiasis samples, and 26 spot were employed for MAILDE-TOF MS/MS analysis. A total of Twenty-one spots were vertified to be protein. including ten up-regulated in tumor samples: Actin,cytoplasmic 2,Annexin A4,Fibrinogen beta chain, Moesin, Tropomyosin alpha-1 chain, eleven up-regulated including Collagen alpha-1(VI) chain, Heat shock protein, Gelsolin, Retinal dehydrogenase 1, Vimentin, Tropomyosin alpha-1 chain ,Actin,aortic smooth muscle, Keratin,type II cytoskeletal 1, Galectin-1 ,Hemoglobin subunit beta ,Superoxide dismutase [Cu-Zn](some protein repeat. RT-PCR and Western blotting show that Moesin, Annexin A4 was up-regulated in cancer adversely Retinal dehydrogenase 1 down-regulated in cancer. Immunohistochemical examination in 59 case gallbladder cancer showed that the enhanced expression of AnnexinA4 was correlated with clinical stage, lymph node involvement and enhanced expression of Moesin was correlated with sex, as well as Liver metastasis Wall invasion, All those being known indicators of a relatively poor prognosis in PCG patients.On the other hand, enhanced expression of Retinal dehydrogenase 1 negative correlation tumor stage and tissue Differentiation. Conclusions:The result suggest that Moesin,AnnexinA4 may be a potential biomarker for diagnosis, prognosis, monitoring in the therapy of PCG, While Retinal dehydrogenase 1 may be a potential prognosis factor.Further, this comprehensive strategy could be used for identifying other differentially expressed proteins that have potential to be candidate biomarkers of Primary carcinoma of the gallbladder.In summary, we for the first time profiled proteome alterations in PCG tissues and these results may provide useful insights for understanding the mechanism involved in the process of Primary carcinoma of the gallbladder.