The Effect Of SiRNA Silencing β-catenin Gene On Human Telomerase Reverse Transcriptase In Gastric Cancer Cells

Objective:To investigate the effect of using RNA interference technology targeting to silence β-catenin gene expression on cell proliferation, division cycle, COX-2 and hTERT in gastric cancer AGS cells.Methods:Using RNA interference technology to transfect the β-catenin-specific small interfering RNA sequence into gastric cancer AGS cells to silence the β-catenin gene expression. Then set the gastric cancer AGS cells into three groups, including the experimental group transfecting with siRNA-β-catenin sequence, the negative control group transfecting with siRNA-control sequence, and the Blank group is normal AGS cells without transfection. Using the fluorescent microscope to detect the transfection efficiency of fluorescence label fragments. Using qRT-PCR to detect the targeting β-catenin gene expression level, as well as the gene expression levels of hTERT and COX-2. Using Western blot to detect the protein expression levels of β-catenin, hTERT and COX-2. Using MTT assay to detect the cell proliferation and using flow cytometry to analyze the cell cycle.Results:Transfect with siRNA specific fragment target to silence β-catenin gene can significantry decrease the expression level of P-catenin mRNA (1.76±0.045 vs 1.00±0.043). Differences are significant between the negative control group and the experimental group (P<0.05). Comparing with the negative control group, hTERT mRNA expression level is significantry decrease (2.37±0.022 vs 1.00±0.030), the difference is significant (P<0.05). Wnt/β-catenin signaling pathway downstream target genes COX-2 mRNA expression level is lower (1.84±0.057 vs 1.00±0.038), comparing with the negative control group the difference is statistically significant (P<0.05). After silencing p-catenin gene, hTERT and COX-2 protein relative expression level is inhibited in the experimental group comparing with the negative control group, and the difference is statistically significant (P<0.05). We find that cell proliferation significantly decreasing at the third day by using MTT assay (P<0.05). We find that silencing β-catenin gene can inhibit cell cycle after 72 hours by using the flow cytometry method. And the cell cycle difference of experimental group is statistically significant comparing with the negative control group (P<0.05).Conclusion:Inhibiting β-catenin gene of AGS cells can reduce the proliferation, interdict the cell cycle and reduce the expression of COX-2. β-catenin gene may become a new target for treatment of gastric cancer. Wnt/β-catenin signaling pathway may be involved in hTERT regulation, and plays some biological function through hTERT and telomerase.