MiR-301a transcriptionally activated by HIF-2αpromotes hypoxiainduced epithelial-mesenchymal transition by targeting TP63 in pancreatic cancer

BACKGROUND Pancreatic cancer(PC)is one of the deadliest cancers worldwide.PC metastasis involves a complex set of events,including epithelial-mesenchymal transition(EMT),that increase tumor cell invasiveness.Recent evidence has shown that hypoxia is a major EMT regulator in pancreatic cancer cells and facilitates metastasis;however,the mechanisms remain elusive.AIM To investigate the role of miR-301a in hypoxia-induced EMT in PC cells.METHODS Real-time PCR and Western blot analysis were used to detect the expression of miR-301a and EMT markers in PDAC cells cultured in hypoxic and normoxic conditions.Western blot analysis was used to detect the expression of EMT markers in PDAC cells with miR-301a overexpression.Wound healing assay and Transwell assay were used to detect the migration capabilities of PDAC cells with miR-301a overexpression and knockout.Luciferase assay was used to detect the miR-301a promoter and the 3’untranslated region activity of TP63.Orthotopic PC mouse models were used to study the role of miR-301a in metastasis of PDAC cells in vivo.In situ hybridization assay was used to detect the expression of miR-301a in PDAC patient samples(adjacent paratumor and paired tumor tissues).RESULTS Hypoxic environment could directly promote the EMT of PC cells.The expression level of miR-301a was increased in a HIF2αdependent manner in hypoxia-cultured CFPAC-1 and BxPC-3 cells.Overexpression of miR-301a enhanced the hypoxia-induced EMT of PC cells,while knocking out miR-301a result in the suppression of hypoxia-induced EMT.TP63 was a direct target of miR-301a and involved in the metastatic process of PC cells.Furthermore,miR-301a upregulation facilitated PDAC distant metastasis and lymph node metastasis in vivo.Additionally,miR-301a overexpression was indicative of aggressive clinicopathological behaviors and poor prognosis.CONCLUSION The newly identified HIF-2α-miR301a-TP63 signaling pathway may play a crucial role in hypoxia-induced EMT in PDAC cells.

Long noncoding RNA negative regulator of antiviral response contributes to pancreatic ductal adenocarcinoma progression via targeting miR-299-3p

BACKGROUND Pancreatic ductal cancer(PDAC)has high malignancy and poor prognosis.Long noncoding RNAs(lncRNAs)are associated with high levels of malignancy,including PDAC.However,the biological and clinical significance of negative regulator of antiviral response(NRAV)in PDAC is unclear.AIM To study the regulatory role of lncRNA NRAV in PDAC.METHODS GEPIA analyzed lncRNA NRAV and miRNA(miR-299-3p)expression levels in PDAC tissues and measured them in PDAC cells by quantitative measurements in real time.The specific role of NRAV and miR-299-3p in cell proliferation and transfer potential was evaluated by cell formation analysis,Cell Counting Kit-8 and Transwell analysis.The relationship between NRAV and miR-299-3p was studied by predictive bioinformatics,RNA immunoassay,and fluorescence enzyme analysis.In vivo experiments included transplantation of simulated tumor cells under naked mice.RESULTS The expression level of lncRNA NRAV was higher in both tumor tissues and cell lines of PDAC and was negatively associated with the clinical survival of PDAC patients.Functionally,overexpression of NRAV promoted cell proliferation and metastasis of PDAC cells,while knockdown of NRAV reversed these effects.Finally,NRAV was performed as a molecular sponge of miR-299-3p.Moreover,overexpression of miR-299-3p could reverse the promoting effects of NRAV on cell proliferation and metastasis of PDAC cells.CONCLUSION NRAV facilitates progression of PDAC as a molecular sponge of miR-299-3p and may be a potential molecular marker for diagnosis and treatment of PDAC.

Genome-wide CRISPR-Cas9 screening identifies that hypoxiainducible factor-1a-induced CBX8 transcription promotes pancreatic cancer progression via IRS1/AKT axis

BACKGROUND Pancreatic cancer(PC)is one of the most lethal malignancies worldwide.It is known that the proliferation of PC cells is a critical process in the disease.Previous studies have failed to identify the key genes associated with PC cell proliferation,using bioinformatic analysis,genome-wide association studies,and candidate gene testing.AIM To investigate the function of the chromobox 8(CBX8)/receptor substrate 1(IRS1)/AKT axis in PC.METHODS A genome-wide CRISPR-Cas9 screening was performed to select genes that could facilitate PC cell proliferation.Quantitative reverse transcription-polymerase chain reaction was used to detect the expression of CBX8 in PC tissues and cells.The regulatory roles of CBX8 in cell proliferation,migration,and invasion were verified by in vivo and in vitro functional assays.RESULTS CBX8 was upregulated in PC tissues and shown to drive PC cell proliferation.Higher expression of CBX8 was correlated with worse outcomes of PC patients from two independent cohorts comprising a total of 116 cases.CBX8 was also proved to serve as a promising therapeutic target for a PC xenograft model.We demonstrated that hypoxia-inducible factor(HIF)-1a induced CBX8 transcription by binding to the promoter of CBX8.CBX8 efficiently activated the PI3K/AKT signaling by upregulating insulin IRS1.CONCLUSION CBX8 is a key gene regulated by HIF-1α,and activates the IRS1/AKT pathway,which suggests that targeting CBX8 may be a promising therapeutic strategy for PC.

Correction to“Genome-wide CRISPR-Cas9 screening identifies that hypoxia-inducible factor-1a-induced CBX8 transcription promotes pancreatic cancer progression via IRS1/AKT axis”

Correction to“Genome-wide CRISPR-Cas9 screening identifies that hypoxiainducible factor-1a-induced CBX8 transcription promotes pancreatic cancer progression via IRS1/AKT axis”(PMID:34853645 PMCID:PMC8603463 DOI:10.4251/wjgo.v13.i11.1709).In this article,the picture of Figure 6C was misused due to our carelessness while typesetting.We corrected this mistake,and replaced the incorrect image with the correct one.