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Effect of DKK1 on proliferation, cell cycle and apoptosis of gastric cancer AGS cells
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Abstract:
Objective：To study the effect of silencing DKK1 (Dickkopf1) gene on the proliferation, cell cycle and apoptosis of gastric cancer AGS cells and the action mechanism. Methods：The DKK1-shRNA vector was constructed and transfected into AGS cells. The stably transfected cell lines were screened. The total protein and RNA of the transfected cells were extracted and the mRNA and protein expressions of DKK1 were detected by qPCR and WB, respectively. The experiment was divided into blank control group (Control),negative control group (shNC) and DKK1 silence group (DKK1-shRNA). CCK8 assay was used to detect the proliferation of AGS cells of each group cultured for 0, 24, 48, 72, 96, 120 and 144 h, and flow cytometry was used to analyze the cell cycle and apoptosis in each group. The relationship between DKK1 and clinicopathological features of gastric cancer was analyzed after searching HPA database.Results：The gastric cancer AGS cells with stable DKK1 gene knockdown was successfully established, and it was confirmed that the mRNA and proteinexpressions of DKK1 in DKK1-shRNA group decreased by 72% and 47%, respectively, compared to shNC group (all P<0.05). The cell proliferation curve showed that, the cell proliferation in DKKl-shRNA group significantly decreased after 72 hour of culture compared with that in control and shNC groups (P<0.05). The cell number of S phase decreased from 32.06% to 25.87%,while the number of G2/M phase increased from 8.49% to 21.26% compared with shNC group (all P<0.05). The number of apoptotic cells also statistically increased from 10.34% to 20.65% (all P<0.05). The data of HPA database showed that DKK1 mRNA level in gastric cancer tissues was significantly higher than that in normal tissues, and the high expression of DKK1 mRNA was negatively correlat‐ed with the survival rate of gastric cancer patients. Conclusion: Silencing DKK1 gene can inhibit the proliferation of gastric cancer cells, arrest cells in G2/M phase and promote cell apoptosis. DKK1 plays a pro-carcinogenic effect in gastric cancer.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No.31560326, No. 31760328,No.31660031), Science and Technology Planning Project of Guizhou Province (No. [2017]5652), and the Science and Technology Planning Project of Guiyang (No. [2017]5-16, [2017]30-4)
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