Objective To investigate the impacts of long non-coding RNA TMPO antisense RNA1(lncRNA TMPO-AS1)on the proliferation,migration and invasion of colorectal cancer(CRC)cells by regulating the miR-340-5p/RUNT related transcription factor 1(RUNX1)axis.Methods The quantitative reverse transcriptase PCR(qRT-PCR)was applied to detect the mRNA levels of TMPO-AS1,miR-340-5p,and RUNX1 in human normal colon epithelial cells HCoEpiC,human CRC cells SW480,HCT116,LOVO,and HT-29.HCT116 cells were transfected with si-ctrl,si-TMPO-AS1,mimics NC,miR-340-5p,si-TMPO-AS1+anti miR-ctrl,and si-TMPO-AS1+anti miR-340-5p.qRT-PCR was applied to detect the mRNA levels of TMPO-AS1,miR-340-5p,and RUNX1 in transfected HCT116 cells.Cell activity,and proliferation were detected by CCK-8 assay and EdU assay,respectively.Transwell assay was applied to detect migration and invasion of HCT116 cells.Western blot was applied to detect the protein expressions of RUNX1,proliferating cell nuclear antigen(PCNA),and matrix metalloprotease-2(MMP-2)in HCT116 cells.Dual-luciferase reporter assay was applied to verify the relationship between TMPO-AS1 and miR-340-5p,and that between miR-340-5p and RUNX1.Results Compared with those of HCoEpiC cells,TMPO-AS1 and RUNX1 were significantly upregulated,and miR-424-5p was significantly downregulated in CRC cell lines(P<0.05).Expression changes in HCT116 cells were the most obvious,and therefore,HCT116 cells were selected for subsequent experiments.Compared with those transfected with si-ctrl,HCT116 cells transfected with si-TMPO-AS1 presented significantly lower mRNA levels of TMPO-AS1 and RUNX1,A value of cell viability,EdU-positive cell rate,numbers of migrating and invading cells,and protein levels of RUNX1,PCNA,and MMP-2,but higher mRNA level of miR-340-5p(P<0.05).Compared with those transfected with mimics NC,HCT116 cells transfected with miR-340-5p presented significantly lower mRNA level of RUNX1,A-value of cell viability,EdU-positive cell rate,numbers of migrating and invading cells,and protein levels of RUNX1,PCNA,and MMP-2,but higher mRNA level of miR-340-5p(P<0.05).Compared with those transfected with si-TMPO-AS1+ anti miR-ctrl,HCT116 cells transfected with si-TMPO-AS1+anti miR-340-5p presented significantly higher mRNA level RUNX1,A-value of cell viability,EdU-positive cell rate,numbers of migrating and invading cells,and protein levels of RUNX1,PCNA,and MMP-2,but lower mRNA level of miR-340-5p(P<0.05).TMPO-AS1 targeted miR-340-5p and negatively regulated its expression,while miR-340-5p targeted RUNX1 and negatively regulated its expression.Conclusion Silencing TMPO-AS1 targets up-regulation of miR-340-5p and upregulates it,thereby down-regulating RUNX1 and inhibiting proliferation,migration,and invasion of HCT116 cells.
long non-coding RNA TMPO antisense RNA1(lncRNA TMPO-AS1)miR-340-5pRUNT related transcription factor 1(RUNX1)colorectal cancer cells
万方数据
维普
