| [Objective] The literature reports that B4GALT3 gene is abnormally expressed in colon cancer,cervical cancer,neuroblastoma and other cells and regulates their biological behavior.In order to investigate whether B4GALT3 can also regulate its biological behavior in gastric cancer cells,the research team confirmed that the gene is highly expressed in BGC823 and SGC7901 gastric cancer cells.Therefore,this study further explores the interference of this gene with siRNA to BGC823 and SGC7901.The biological behavior of two gastric cancer cells provides a theoretical basis for the clinical treatment of gastric cancer in the future.[Methods] 1.Design and synthesize three pairs of B4GALT3 siRNAs;2.Western Blot to screen down the most significant pair of siRNAs and concentrations of B4GALT3 gene from three pairs of B4GALT3 siRNAs;3.Transiently transfect the selected B4GALT3 siRNA into BGC823 And SGC7901 two gastric cancer cells,and the migration and invasion abilities of two gastric cancer cells BGC823 and SGC7901 were detected by scratch testand transwell migration assay.The proliferation activities of two gastric cancer cells BGC823 and SGC7901 were detected by MTT assay.4.Western Blot examine the effect of down-regulating the B4GALT3 gene on the expression of EMT-related proteins such as E-cadherin.[Results] 1.Three pairs of siRNAs were successfully constructed.2.Western Blot screening results showed that B4GALT3 siRNAs interfered with B4GALT3 successfully,GGAGGAGTCTCAGCACTTA was the best interference sequence,and the optimal interference concentration was 8μM.3.The results of scratch test showed that after B4GALT3 gene down-regulation,the migration distance of BGC823 and SGC7901 cells decreased,P<0.05,the difference was statistically significant,indicating that down-regulation of B4GALT3 gene can inhibit the migration rate of these two gastric cancer cells.Transwell migration assay showed that after B4GALT3 gene down-regulation,the migration ability of BGC823 and SGC7901 gastric cancer cells decreased,P<0.05,the difference was statistically significant,indicating that down-regulation of B4GALT3 gene can inhibit the migration ability of these two gastric cancer cells;Transwell invasion results showed that the invasive ability of BGC823 and SGC7901 gastric cancer cells decreased after B4GALT3 gene down-regulation,P<0.05,the difference was statistically significant,indicating that down-regulation of B4GALT3 gene can inhibit the invasion ability of these two gastric cancer cells.4.The results of MTT assay showed that the proliferation of BGC823 and SGC7901 cells was decreasedafter B4GALT3 gene down-regulation,P<0.05.The difference was statistically significant,indicating that down-regulation of B4GALT3 gene could inhibit the proliferation of these two gastric cancer cells.5.The results of EMT showed that the expression of EMT-related protein in BGC823 and SGC7901 gastric cancer cells was down-regulated after B4GALT3 gene expression: transfection group relative to control group and siCTL group Claudin-1 protein and E-cadherin protein expression level was significantly decreased,P<0.05,the difference was statistically significant.The protein expression levels of Vimentin,Slug and β-catenin were significantly increased,P<0.05,the difference was statistically significant.The results showed that down-regulation of B4GALT3 gene expression could affect EMT in two gastric cancer cells.[Conclusion] The expression of B4GALT3 gene by siRNA can inhibit the migration,invasion and proliferation of gastric cancer cells BGC823 and SGC7901,and the specific mechanism may be related to EMT. |
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