The Effect And Mechanisms Of LncRNA SNHG16 On Proliferation,invasion And Apoptosis Of Gastric Cancer AGS Cells

Objective:Knockdown of SNHG16 by RNA interference in vitro to indicate the effect of cell proliferation,migration,invasion,apoptosis and cell cycle and the potential mechanism in gastric cancer AGS cells,and to explore the potential possibilities as a target for the treatment and prognosis of gastric cancer.Methods:?1?Silencing of SNHG16 was performed by Lipofectamine?RNAiMAX transfection,and the expression of SNHG16 was detected by Quantitative Reverse Transcription PCR?qRT-PCR?.?2?The cell viability,proliferation,invasion and migration of gastric cancer AGS cells after SNHG16 knockdown were determined by CCK-8assay,clone formation assay,wound healing assay and transwell assay,respectively.Survival rate of AGS cells after combined use of 5-Fu was perfomed by CCK-8 assay.The cell cycle and apoptosis were detected by flow cytometry and the morphology of cell apoptosis was observed by Hoechst 33342 staining.?3?The relative expression of cyclinD,cyclinA,cyclinE,cdk2,cdk6,cdc25A,c-myc,PCNA,p27,p21 and p53 were were revealed by western blot,respectively.Results:?1?SNHG16 was successfully silenced in AGS cells,the interference efficiency of si-SNHG16 was?70.3±6.5%?%,and the transfection efficiency observed by fluorescence microscope was up to90±2.56%.?2?The CCK8 assay results show that the cells viability percentage of AGS were decreased after knocking down SNHG16,and the cells viability percentage of si-SNHG16 group was further decreased when combined with 5-Fu.Clone formation assay showed that the proliferation of AGS cells was inhibited after knockdown of SNHG16.The expression of PCNA was also significantly reduced.?P<0.01??3?Wound healing assay and transwell assay results showed that the migrant and invasive ability of AGS was weakened after knockdown of SNHG16.?4?The results of Hoechst33342 staining and flow cytometry showed that apoptotic cells and apoptosis rate of AGS cells were increased after knockdown of SNHG16.?5?The results of flow cytometry showed that after knockdown of SNHG16,the proportion of G₀/G₁ phase was increased,the proportion of S phase was decreased,G₂/M phase was no difference,and cells were arrested in G₀/G₁ phase.?6?Western blot assay indicated that SNHG16 knockdown resulted in upregulated expression of p21 caused by the high expression of p53 and decreased cdc25A and up-regulated p27 when the expression of c-myc was suppressed respectively in AGS cells,and then ultimately leaded to the suppression of cdk2,cdk6,cyclinA,cyclinE and cyclinD.Finally silencing SNHG16 resulted in accumulation of G₀/G₁ phase cells and inhibited the cell proliferation.Conclusion:?1?LncRNA-SNHG16 can promote the cell proliferatio-n,migration,invasion,apoptosis and cell cycle progression of Gastric cancer cell AGS.?2?SNHG16 plays an important role as regulator in up-regulating the expression of c-myc and down-regulating of p53.Over expression SNHG16 may accelerate the progression of cell cycle and promote the proliferation of gastric cancer by enhancing c-myc expression and inhibiting the expression of p53,which devote to the development of gastric cancer.?3?SNHG16 may be involved in the development of gastric cancer as an oncogene and is expected to be a molecular target for the treatment and prognosis of gastric cancer.