Mechanism of miR-210 involved in epithelial–mesenchymal transition of pancreatic cancer cells under hypoxia

Abstract

Purpose: To investigate the possible mechanism of miR-210 involved in epithelial–mesenchymal transition (EMT) of pancreatic cancer cells under hypoxia.

Methods: In this study, we used the following approaches. Hypoxic microenvironment was stimulated in vitro, and the CCK-8 assay was used to analyze cell viability. The MiRNA expression level was measured by qRT-PCR. HOXA9, EMT-related proteins, and NF-κB activities were examined by immunoblotting assay. Dual luciferase reporter assay was used to assess whether HOXA9 was a target of miR-210.

Results: Under hypoxia condition, miR-210, HIF-1α and NF-κB were increased, and the HOXA9 was reduced in PANC-1 cells. When miR-210 was overexpressed in normoxic PANC-1 cells, EMT epithelial markers of E-cadherin and β-catenin were down-regulated, and mesenchymal markers of vimentin and N-cadherin were up-regulated to promote cell migration/invasive ability, and the HOXA9 level was decreased. After HOXA9 level decreased, the sensitivity to chemotherapeutic drug of gemcitabine was reduced, NF-κB expression level and cell migration/invasive ability was enhanced. Whereas, miR-210 antagonist into hypoxic PANC-1 cells, which up-regulated E-cadherin, β-catenin level, and down-regulated vimentin and N-cadherin levels to decrease cell migration/invasive ability, and increase the HOXA9. Furthermore, increasing HOXA9 level decreased NF-κB expression level and cell migration/invasive ability, enhanced the sensitivity to gemcitabine. At last, miRDB and TargetScan predicted that HOXA9 was a target of miR-210, and dual luciferase reporter assay verified this hypothesis.

Conclusion: MiR-210 inhibited the expression of HOXA9 to activate the NF-κB signaling pathway and mediated the occurrence of EMT of pancreatic cancer cells induced by HIF-1α under hypoxia.

Keywords:

• Pancreatic cancer cells
• hypoxia
• miR-210
• nuclear factor κB
• hypoxia-inducible factor-1α

Acknowledgments

The authors thank Dr Li-Ping Cao and his research team for direction and support regarding experimental techniques. The authors also thank Rebecca Porter, PhD, and Mitchell Arico from Liwen Bianji, Edanz Editing China (www.liwenbianji.cn/ac), for editing the English text of a draft of this manuscript.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors.

Disclosure statement

The authors declared no conflict of interest.

Additional information

Funding

The authors received financial support from the Zhejiang Health Science and Technology Talents Project.