| BackgroundGastric cancer is currently the highest incidence of malignant digestive system tumors.The onset of gastric cancer is insidious.It often lacks specific symptoms,signs and early diagnosis indicators in the early stage.Therefore,it is used worldwide,especially in developing countries and less developed countries or regions,especially In the absence of local people's awareness of regular physical examination,gastric cancer is often misdiagnosed as peptic ulcer,chronic gastritis,Crohn's disease and other digestive diseases,thus delaying the important timing of gastric cancer treatment.This research group has long been committed to the reseach on regulatory role of the membrane rivet repeat sequence protein 6Ankrd6(diversin)in tumorigenesis and development of tumor tissues and cells and its molecular mechanism.it is shown in previous study that the central domain of Diversin can be combined with seious of signaling molecules and key proteins in celluler signaling pathways to play a role of regulatory in tumorigenesis and development.In addition to the abnormal expression of tumor genes at the genesis and development of tumors,post-translational modification of proteins can affect the function of more than half of the proteins.And it is illustrated in previous studies that sialylation of proteins plays an important role in the occurrence and development of gastric cancer cells.We collected tissue wax blocks from gastroscopy specimens of 77 gastric cancer patients from the Central Hospital Affiliated to Shenyang Medical College in the early stage,and evaluated the expression level of Diversin by immunohistochemistry.The results showed that the expression level of Diversin in gastric cancer tissues was significantly higher than that in adjacent tissues.The expression of Diversin is related to gastric cancer tumor diameter,TNM stage,lymph node metastasis and other factors,which provides an important preliminary basis for the development of this experiment.MethodsDiversin overexpression plasmids carrying and Diversin sh RNA interference plasmids carrying neomycin and puromycin resistance repectively have been constructed.We screened the screening concentrations of gastric undifferentiated cancer cells HGC27 and gastric differentiated cells MNK45 in G418 and puromycin.Using the above plasmids to transfect HGC27 cells and MNK45 cells and under the conditions of the G418 concentration medium previously screened Diversin-overexpressing cells were cultured,and gastric cancer cell lines down-regulated by Diversin were cultured in a medium at an appropriate puromycin concentration to obtain gastric cancer cell lines that were more up-regulated and down-regulated.Then MTS,transwell,wound healing,and clone formation were used to evaluate the capablity of proliferation,invasion,migration,and malignancy transformation of gastric cancer.To further explore the function of Diversin,lectin blot was used to check the level of glycosylation,western blot were uesd to evaluate the expression level of pryoptosis and autophagy of HGC27 cell line.ResultsMTS cell proliferation experiments show that Diversin can up-regulate the proliferation of HGC27 and MNK45 cells in the stomach;clone formation test results show that Diversin can up-regulate gastric undifferentiated cancer cells HGC27 cells and gastric differentiated cancer cells MNK45 Malignant transformation ability of cells;Transwell results show that Diversin can up-regulate the invasion and invasion of gastric undifferentiated cancer cells HGC27 cells and gastric differentiated cancer cell MNK45 cells;agglutination The results of the Western blot test showed that Diversin can increase the level of ?2,6 sialylation in the proteome of gastric undifferentiated cancer cells HGC27 cells.ConclusionDiversin promotes the proliferation,invasion and metastasis of gastric cancer cells;Diversin can increase the level of ?2,6 sialylationthe of proteome of gastric undifferentiated cancer cells HGC27 cells... |
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