Study On Relevant Proteome Of Human Gastric Carcinoma

The incidence of stomach cancer stands the second position in the malignant tumor in the word and the first in china. Its process is very complex . Scores of factors make their roles in it .According to the research of modern molecule biology , human Genome makes up of about 20,000 or 25,000 genes . They all have their specific properties during the process of monitoring and regulating the origination and expandance of tumor cells. So,the research on rous of gene in gastric caranoma tissue is very important .It can help to make cancer mechanism clear and offer the theorial basis to diagnosing and treatment of stomach cancer . We sieve the discrepancy proteins and research their characters by means of Proteome analysis technique. In this research, We contrast the outcomes of lineage analysis to immunity histochemistry [IHC] and try to find the key or "on-off "gene on the base of gene arrangement and location. In the first part of our study, we verify the association between Her-2, bcl-2, p53, VEGF, MMP-2 and clinical characters of gastric carcinoma by IHC. Moreover, we examine the serum level of ICAM-1 among the relevant masses and analyse the relationship between ICAM-1 and invasion of stomach cancer cells. In the second part ,we sieve the particular protein from normal and tumor tissue and find the specific sites of discrepancy protein . We choose ten sites from them and identificate them by MALDI-TOF. Meanwhile, we probe the discrepancy protein by means of data-bank retrieval. As our conclusion , We find that the positive ratio of Her-2, bcl-2, p53,VEGF and MMp-2 is associated with the invasion of tumour cells respectly (P < 0.05). The positive expression of Pl85which acts as the protein coded by Her-2 is distinctly different in poor differentiation group (79.2%) from in high differentiation one (42.1%). In addition, there is no statistical significance of bcl-2 which vary between clinical stage Ⅲ+ Ⅳ(74.1%)or poor differentiation group(75%)and stage Ⅰ+ Ⅱ(68.8%) or high differentiation one (68.4%). The positive ratio of p53 is much higher in stageⅢ+Ⅳgroup(70.4%) than in stage Ⅰ+Ⅱ(31.3%).VEGF is associated with tumor cell's differentiation and invision . whose positive ratio is higher in poor differentiation group (62.5%)than in high differentiation one (36.8%). Moreover, the positive ratio of MMP-2 is also different between high differentiation group(57.9%) or stage Ⅲ+Ⅳgroup (85.2%) and poor differentiation one (87.5%) or stage Ⅰ+Ⅱ(56.3%). The positive ratio of VEGF in gastric carcinoma where p53 express positively is 66.7% nd 31.6% in negative one(P<0.05). The positive ratio of VEGF is much higher in diffuse-pattern group than in other groups(P<0.05). MVC shoot its data upto 76.5±14.7 in diffuse group.The variation of them have its statistical significance (p<0.05) Immunohistochemical staining and FACS were used in this study to detect the level of ICAM-1 expression in tissue and peripheral blood of various group. ICAM-1 was not observed in 20 gastric ulcer tissue, in spite of significantly infiltration of lymphocyte. ICAM-1 was expression in both adenocarcinoma and mucoid cytocarcinoma, among which the expression level of highly differentiated adenocarcinoma was higher than less differentiated adenocarcinoma (P<0.01). It's not significantly different between Adenocarcinoma and mucoid cytocarcinoma (P > 0.05). Twodistinct cellular distributions of ICAM-1 were observed, either diffuse cytoplasmic staining and /or expression on the cell membrane, both patterns could be found in the same tumor. sICAM-1 was detected in serum in all of gastric cancer group and control group, when we detected sICAM-1 in various group, the level of sICAM-1 was higher in the serum of patients with gastric cancer than normal group (P<0.01), liver metastatic cancer group higher than non-liver metastatic cancer group (P<0.01), it's not significantly different between less differentiated adenocarcinoma and highly differentiated adenocarcinoma, between adenocarcinoma and mucoid cytocarcinoma.(P>0.05). After sieving the protein from 4 gastric carcinoma specimens and 2 normal mucous membrane samples, We analyse the data by Imagemaster 2D platinum analysis software. We verify 1,147 protein spots in gastric carcinoma tissue and 1,079 ones in normal specimens on the formed glue-plate. Among them, there are 164 discrepancy protein spots. We find that 41 spots only emerge in gastric carcinoma and 27 only in normal mucous. In our study, we verify 9 discrepancy spots in gastric carcinoma and 1 spot in normal mucous by MALDI-TOF. Finally, we confirm 8 frequently shown proteins by data-bank analysis. They are Chain F, Cypa Complexed With Hvgpia;peptidylprolyl isomerase A;proapo-A-I protein;Chain B, Crystal Structure Of The Human Cu, Zn Superoxide Dismutase;similar to Cofilin, non-muscle isoform (Cofilin-1);protein PP4-X;pyruvate dehydrogenase E1-beta subunit precursor; Drug-Protein Interactions: Structure Of Sulfonamide Drug Complexed With Human Carbonic Anhydrase。In recent years, some new techniques are used in the research of stomach cancer such as contrast gene group hybridize andmicro-array. But there are amount of falt positive signals during the process. In the other side, the alteration of DNA or mRNA probably confine the function of them, too . The study of protein group probably make up the above two weakness. The results from IHC show that Pl85, VEGF and MMP-2 are obviously associated with pathologic classification and clinical stage. So, we maybe evaluate the process of gastric carcinoma by means of verifying their positive ratio .The extra-expression of bcl-2 mainly have its role during initial stage of tumors. In the meanwhile , We find that p53 is closely associated with pathologic classification though there is no relationship between p53 and clinical stage of gastric carcinoma. Furthermore, as a key protein, p53 maybe up-regulate the expression of VEGF and urge the proass of angiogenesis. At the second stage of our study , we find that 8 specimens from the verified 10 protean spots are different from normal ones reported in the newly publications. We can infer that protein-group-analysis technique maybe offer a new way to severing the specific markers of gastric carcinoma and lay a basis for explaining the mechanism of tumor by gene research. Nowadays, the relevant study on proteome of gastric carcinoma is rarely reported in the well-known publications. It's at the initial stage in the domestic research yet . We take the lead in learning about discrepancy protein by way of 2-DG and computer-graph analysis. Meanwhile,We analyse the datum of IHC and 2-DG.. Finally,8 specific protein specimens are verified. Particularly, these proteins are different from known ones. It maybe explain the limitness of studying on gene-group. In the other hand , the way of proteome study offer a good idea to sever the specific markers of...