| Background and ObjectiveGastric cancer is a malignant neoplasm which has a high in incidence and is prone to invasive local organ and metastasis. Though the comprehensive treatments including surgery has been used in gastric cancer,advanced gastric cancer patients have poor prognosis. So find some biological markers to diagnose and predict the progress become important, our research are designed to study the expression of C4.4A in gastric cancer and surround normal gastric issues,analyze the correlation between the expression and the clinical date in order to Provide a theoretical basis to explore its mechanism in gastric cancinogenesis of furtherly prevention and clinical treatmentMethodsClinical date and tissue construction:All the specimens of121urgically resected patients of gastric cancer,121cases of paried normal gastric mucosa(>5cm Away from the edge of the tumour) were collected from Huashan Hospital of Fudan University, one small piece of tissue was put into the nitrogen canister and then stored in-80℃refrigerator, the rest tissue were immersed in10%Formalin solution.then the gastric cancer tissues were cut into4μm sections to performance Immunohistochemical staining to detect the expression of C4.4A protein Immunohistochemical method andassement:S-P method was used to detect the expression of C4.4A in gastric cancer and gastric mucosa.Staining C4.4A was located both in the nucleus and cytoplasm/two independent observers who were blind to the message of the patients evaluated the outcome of C4.4A staining in200cells from two selected representative fields. the staining intensity was classified as0(negative),1(weak),2(moderate).and3(strong).the percentage of positive cells that in200cells were divided into0(<5%),1(5%-25%),2(26%-50%) and3(>50%)。the resulte of the staining was the product of the two aspects. RT-PCR:According to the kit instruction, respectively, extracting the total mRNA in gastric cancer and normal mucosa tissue, using apectrophometer measurement and quantitative purity, usinging the total mRNA as the first cDNA chain, adding according reaction system of the substrate and enzyme, after cDNA preparation joining the PCR amplification, DNA amplification, decting the amplification of CT valuesResult:positive C4.4A staining was found mainly in Cell membrane and in Cytoplasm of gastric cancer and metastatic lymph nodes and it is not found in normal surround gastric tissue,C4.4AmRNA expression was significantly up-regulated in gastric cancer as compared with that in normal tissues(p<0.05),88.4%cases had positive staining of C4.4A by immunohistochemistry.the depth dying of C4.4A is associated with the size,depth of invasion,lymh node metestasis,distant metastasis,the TNM stage of gastric cancer,it has no relationship with patients’s sex, age, histological types,Moreover,Kaplan-Meier analsis showed that C4.4Aoverexpression was corresponding a worse survival (p<0.05),multivariable analysis indicated that C4.4Aexpression was an independent prognostic indicator for gastric cancerConclusion:These results suggest a role for C4.4A in the incidence and progress of gastric cancer the potential increase of C4.4A expression in the development, invasion, and metastasis of gastric cancer... |
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