Study Of The Expression Of Fascin In Gastric Cancer And Study Of Prognosis-Realted Proteomics In Advanced Gastric Cancer

Objective:Fascin,a structurally unique actin cross-linking protein,which functions in the organization of two major forms of actin-based strctures:cell protrusions and cytoplasmic microfilament bundles.Cell protrusions include filopodia,spikes, microvilli and dendrites of dendritic cells,which have roles in cell-matrix adhesion, cell interactions and cell migration.While cytoplasmic bundles participate in the formation of cell architecture.Recent data showed that fascin is up-regualted in some epithelial carcinomas and correlates with the poor survival.But the expression of fascin in the gastric carcinoma is still not known much.Methods:Paraffin sections of gastric carcinoma of 249 patients were investigated by immunohistochemistry with monoclonal antifascin antibody,those that were stained more than 5%of tumor cells were recorded as positive immuno reactivity.Results:In 249 patients,55 were found positive fascin immunoreactivity(22.1%),37 were classified "1+"(14.9%),and 18 of the patients were classified "2+"(7.2%). Furthermore,with investigating the relationships between fascin expression and clinicopathological characteristics,we found the expression of fascin correlated with the patient ages(p=0.032),and the differentiation degree of the gastric carcinoma.The expression of fascin was significantly higher in well differentiated carcinoma than those in poorly-differentiated group(p=0.001).while no correlations were found between the sex,T stage,N stage,M stage and the tumor location).Those patients with no fascin expression seemed to have a better prognosis than those patients with fascin(2+) expression.Conclusion:1.There is no expression of fascin in normal stomach epithelium,but there is fascin expression in gastric cancer,which indicated that the change of skeleton of gastric carcinoma ceils.2.There are different fascin expression in different differentiated gastric carcinoma. Which indicated that the expression of fascin is correlated with the tumor differentiation.3.Those patients with fascin expression may have a poor prognosis. [Objective]Gastric cancer is one of the main malignant tumors in our country,and it has certain morbility and mortility.Although the 5-year survival rate of early stage gastric cancer can proceed about 85%,but the number of these patitent is just only 10% to 30%.Most of the patients with gastrc cancer are found to be late stage,so the prognosis of these patients are poor,and different from each other.So,it is important to find some markers to staging the tumor stage correctly,and to judge the real prognosis of the advanced gastric cancer.A tumor marker is a substance found in the blood,urine,or body tissues that can be elevated in cancer,among other tissue types.It can be produced by the tumor cell or by the reaction of the body,it can relect the existence and the growth of the tumor.Tumor marker can be used as the detection,diagnosis,treatment,surveillance and the judgement of the prognosis.CEA and CA19-9 are the markers used now in clinic. Because of the low sensitivity and specificity,these markers are mostly be used as the prescreening and the surveillance for the patients with gastric cancers.The use of these tumor marekers are limited in the judgement of the prognosis of the patients with gastric cancers.There are still no tumor markers that could be used to judge the prognosis of the advanced gastric cancer.Proteomics,a newly technique used to analysis the cellular protein composition, expression and modification,is very useful for identifying disease-associated proteins that show changes in expression and modification corresponding to a disease condition. And with rapid development of technique of Mass Spectrum and 2-DE,it si possible to find out some tumor-realted proteins as the tumor mareker to judge the stage and the prognosis of the patients with gastric cancer.These tumor-related proteins are expected to be biomarkers for diagnosis and putative targeted proteins for treatment[Methods]In the study,to identify the prognosis related proteins of advanced gastric cancer with different prognosis,we selected two groups of cancer tissues from stage Ⅲ～Ⅳgastric cancer patients with very different survival time for study(For good survival group,the average survival time is＞34 months,and for poor survival group, the average survival time is＜15 months.) Nanoflow liquid chromatography system interfaced with a liner ion trap LTQ mass spectrometer was used and database searches were done against NCBI non-redundant database and SWISS-PROT database on an in-house Mascot server and the identified proteins were classified through an online "Web Gene Ontology Annotation Plot" tool.With so many different proteins,we choose two obviously different proteins S100P and HnRNP A2/B1.With the method of Immunohistochemistry,we detect the expression of S100P and HnRNP A2/B1 in 93 patients with advanced gastric cancer.Furthermore,we also analyse the result with the clinicopathological parameters with SPSS 13.0.[Results]With the method of proteomics,There were 284 and 213 proteins identified for group G and group P Prespectively.Among those,130 proteins were detected exclusively in group G and 58 proteins exclusively in group P.There were 14 proteins whose fold change is more than 3 times and 28 proteins whose fold change is more than 2 times.These protein markers function in calcium ion signaling pathway,cellular metabolism,cytoskeleton formation,stress reaction,etc.We found that in these two groups,the difference lies in numbers but not categories and several of these proteins were verified to be related with prognosis or the metastasis of gastric cancer patients.With the statitic analysis of the two group of advanced gastric cancer with different prognosis.We found that the status of metastatic(＜N1 vs＞N1) lymph node is predominant in the prognosis of advanced gastric cancer.So we detect two different protein S100P and HnRNP A2/B1.We found the expression of S100P is about 47.3,and the expression is closely related with prognosis P=0.0375),also the expression is related with the depth of tumor invasion and the status of lymph node metastasis(＜N1 vs＞N1)(P = 0.006,P = 0.006).While the expression of HnRNP A2/B1 is about 92.6%.and there is no significant with the expression and teh prognosis.Furthermore, we also detect the expression of HnRNP A2/B1 in the paratumor tissue.[Conclusion]1.With the method of nanoflow liquid chromatography tandem mass spectrometric approach,we can select the different expressed-protein in the advanced gastric cancer patients with different prognosis.These protein markers function in calcium ion signaling pathway,cellular metabolism,cytoskeleton formation,stress reaction,etc. These can be provide the basis of tumor markers to judge the stage and the prognosis.2.The status of lymph metastasis(lymph node exceed the N1) is a key factor to jude the prognosis of advanced gastric cancer.3.S100P can reflect the status of lymph nodes in patients with advanced gastric cancer.So it can be a tumor marker to judge the prognosis for patients with advanced gastric cancer.4.HnRNP A2/B1 is not a ideal marekr to judge the prognosis for patients with advanced gastric cancer.5.The proteins selected by proteomics to reflect the prognosis need to be verified further.In all,the nanoflow liquid chromatography tandem mass spectrometric approach is efficient in identifying differentially expressed proteins.With the identification of some protein selected,we know that the method is relaible and need to be further verified.Using proteomic method,several prognosis-related proteins were identified and it provide a basis for the further reasearch.