| Gastric carcinoma(GC),mainly referred to as gastric adenocarcinoma,is more common in malignant tumors of the upper gastrointestinal tract.According to China’s epidemiological statistics from 2009 to 2011,the incidence and mortality of gastric cancer in the national malignant tumors are all ranked second.Although the clinical manifestations of early gastric cancer are not characteristic,the diagnostic probability of early gastric cancer is significantly increased with the enhancement of people’s health awareness and the current level of endoscopy and diagnosis,Which not only prolongs the survival time of gastric cancer patients,but also improves the quality of life of patients with gastric cancer.Moreover,under the impetus of precision medicine,the prevention of gastric cancer from the source-molecμle level of gastric cancer has become a top priority in current scientific research.Under the influence of some carcinogenic factors,The stability of the genome is weakened:the oncogene activity is enhanced,the activity of tumor suppressor genes is weakened,and the tumor susceptibility gene is mutated.All of these can cause changes in the protein expression profile of the cell and further induce tumorigenesis.The role of micro RNAs in the development of tumors is obvious,Micro RNAs play an important role in the development of tumors.The principle of their function and the mechanism of regμlating target genes have been confirmed by numerous studies.β-1,4-galactosyltransferase Ⅲ(B4GALT3)is the third member of the β-1,4-galactosyltransferase family,which is involved in the formation of poly-N-acetylgalactosamine,which is closely related to tumorigenesis.Up to now,the expression of B4GALT3 in gastric cancer cells and its influence on the biological behavior of gastric cancer cells are still unknown.Objective:To investigate the effect of mi R-27a-3p on the expression of B4GALT3 and the effects of mi R-27a-3p on the biological behavior of gastric cancer BGC-823 cell,which to lay a foundation for the further study of the effects of B4GALT3 and glycosylation on the biological behavior of gastric cancer.Methods: Western blot was used to detect the expression of B4GALT3 protein in gastric cancer cell lines——SGC-7901(Medium differentiation),MGC-803(low differentiation),BGC-823(low differentiation)and normal gastric mucosa cells(GES-1).Bioinformatics methods were used to obtain mi RNAs that specifically bind to B4GALT3.Western blot was used to study the expression of B4GALT3 in gastric cancer through up-/ down-regulation of mi R-27a-3p levels in BGC-823 cells by transient transfection,Then preliminary analysis of the relationship between the mi R-27a-3p and B4GALT3.The effects of mi R-27a-3p on the biological behaviors of gastric cancer BGC-823 cells were observed using MTT,wound healing,Transwell migration and invasion,and Western blot.Results: Western blot showed that the expression of B4GALT3 in gastric cancer SGC-7901,MGC-803 and BGC-823 cells was significantly high expression than GES-1 cells,P<0.05.mi R-27 a,mi R-27 b,mi R-338-3p,mi R-125 b,mi R-28-5p,mi R-379 and mi R-491-5p et.al seven mi RNAs were identified by bioinformatics methods.Combined with the literature,we selected mi R-27 a for follow-up studies.In addition,It is found that mi R-27a-3p binds to 213~220 positions of the 3 ’UTR of B4GALT3,and the binding sequences are 5’-ACUGUGA-3’,so mi R-27a-3p was selected for further study.Western blot showed that the expression of B4GALT3 protein was decreased after mi R-27a-3p was up-regulated,while the expression of B4GALT3 protein was increased after mi R-27a-3p was inhibited,P<0.05.The results of MTT assay showed that the proliferation activity of mi R-27a-3p cells was significantly enhanced in BGC-823 cells,and the effect was more obvious at 72 h~96 h.On the contrary,after mi R-27a-3p was inhibited,cell proliferation was observed.The activity was significantly decreased,P<0.05.Wound healing experiments showed that mi R-27a-3p mimic group migration distance is higher than the control group,mi R-27a-3p inhibited migration distance was significantly lower than the control group,P<0.05.Trasnwell migration experiments showed that the mi R-27a-3p mimic group migrated through the cell significantly more than the control group;inhibit mi R-27a-3p,the number of migrating cells was significantly reduced,P<0.05.Trasnwell invasion experiments showed that the number of cells invaded by mi R-27a-3p mimic group was greater than that in the control group after invading the BD Matrigel cell,while the number of cells after mi R-27a-3p was inhibited was less than that in the control group,P<0.05.Western blot results showed that mi R-27a-3p mimic could down-regulate the expression of E-cadherin and Claudin-1 and increase the expression of Vimentin,Slug and β-catenin(P<0.05).On the contrary,after mi R-27a-3p was inhibited,The expression of E-cadherin and Claudin-1 was up-regulated while the expression of Vimentin,Slug,and β-catenin were down-regulated(P < 0.05).Conclusion: mi R-27a-3p may promote the proliferation activity,migration and invasion and EMT progression of BGC-823 cells by inhibiting the expression of B4GALT3. |
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