Polypeptide: N-acetylgalactosaminytransferases 2's Role During Adhesion And Migration Of Gastric Cancer Cell SGC7901

Objective: The initiation step of mucin-type O-glycosylation is controlled by a large family of homologous N-acetylgalactosaminyltrans- ferases (ppGalNAcT) which transfer GalNAc to serine and threonine acceptor sites .Their activity is closely concerned with tumor. Invasion and migration are essential during tumor growth and development. Invasion is internal agent of tumor metastasis. During tumor cell invasion and metastasis it is committed step that how to pass through biological containment. In the process, tumor cell adhere basement membrane or extracellular matrix , then release and stimulate many proteolytic enzyme to degradation basement membrane or extracellular matrix and migration to target tissue with help of chemotactic factor,autocrine and paracrine growth factor, accordingly emerge metastasis by cloning and proliferation. This thesis aims to approach the function of ppGalNAcT2 during invasion and migration of gastric cancer cell .Methods:Transfected ppGalNAc-T2 plus sense vecto(rpEGFP-C1-T2),vacuity vector(pEGFP-C1)and interference vector (pSilenCircle) to SGC7901. We detected expression of tumor infiltration correlated molecule (TGF-β1,MMP2,MMP14)by RT-PCR and Western-blot. MTT detected the role of ppGalNAcT2 during its proliferation. Flow cytometer detected cell cycle. 2. Took fibronectin,matrigel and hyaluronic acid to detected the adhesive attraction between different subtype SGC7901 and extracellular matrix (ECM). Utilized Transwell cabin, paved matrigel on the film and constructed recombination basement membrane that fibronectin was chemotactic factor. Experimental animal was nude mouse, masculinity, 4-6 week.Twenty nude mouses were divided randomly into 4 grops. Took different subtype SGC7901 which was occupied exponential phase of growth ft. cell suspension . Carried out subcutaneous inoculation in right armpit by the concentration of 1×107/200μL. In order to make the model of human gastric carcinoma transcellular animal. Statistics analyzed the rate of nude mouse growing tumor and the weight of the tumor. Carried out HE staining to tumor tissue.The result: ppGalNAc-T2 has negative correlation with TGF-β1,MMP2,but no manifest correlation with mmp14 detected by RT-PCR,also by Western-blot. ppGalNAc-T2's high expression suppressor cell proliferation detected by MTT and Flow cytometer .Contrast to SGC7901 which interference vector, the one transfected ppGalNAc-T2 plus sense vector had low adhesion capability to Matrigel, Hyaluronic Acid and Fibronectin, but alike migration capability。The rate of nude mouse growing tumor had no statistics difference(P>0.05), but the weight of tumor had(P<0.05).Conclusion: It is possible that ppGalNAc-T2's low expression and adhesion and migration of gastric cancer have correlation.