The Effect Of CircALPL Sponges MiR-127 By Competing With MTDH On Gastric Cancer Cells

Objective:To explore the specific mechanism of circ ALPL on gastric cancer cells by competitive binding mi R-127.Methods: In this study,we chose circALPL for further research;q RT-PCR was used to detect the abundance of target molecules in the isolated cell nucleus and cytoplasm part;bioinformatics was used to predict the binding micro RNA of circ ALPL and the target gene of micro RNA.The expression level of mi R-127 and MTDH in gastric cancer cells was further detected by q RT-PCR;luciferase reporter gene experiment and RNA immunocoprecipitation technology were used to verify the binding relationship between circ ALPL-mi R-127-MTDH.RNA interference technology inhibited the expression of circ ALPL.Western blot and q RT-PCR were used to detect the transcriptional and post transcriptional changes of genes after circ ALPL inhibition.Results: isolating nuclear and cytoplasmic part,it was found that circ ALPL was mainly distributed in the cytoplasm.Bioinformatics method predicted that there was mi R-127 binding site in circ ALPL.The expression of mi R-127-5p generally decreased in gastric cancer cell lines.Luciferase report gene experiment and RIP test confirmed that circ ALPL could bind to mi R-127;targetscan was used to predict the target gene of mi R-127,and it found that there was a binding site between mi R-127 and MTDH;MTDH was up-regulated in gastric cancer cells,and mi R-127 could bind to MTDH confirmed by luciferase reporter gene test,which indicated that MTDH could be regulated by mi R-127.Overexpression mi R-127 can inhibit MTDH expression.Rip experiment result showed that circ ALPL,MTDH and mi R-127 were enriched in ago 2,indicating that circ ALPL and MTDH could competitively combine with mi R-127 to form ago 2 related silencing complex.The knockout of circ ALPL can reduce the enrichment of circ ALPL on ago 2,but increase the enrichment of MTDH on ago 2.Inhibition of circ ALPL can reduce the expression of MTDH.These results indicate that circ ALPL acts as ce RNA and competes with MTDH to bind mi R-127.Conclusion:1.circALPL competes with MTDH to bind miR-127.2.Interfering with circALPL increases the binding of mi R-127 to MTDH and inhibits the expression of MTDH.