A novel upregulated hsa_circ_0032746 regulates the oncogenesis of esophageal squamous cell carcinoma by regulating miR-4270/MCM3 axis

Sep 21, 2026

Uploaded by Jiven

A novel upregulated circular RNA, hsa_circ_0032746, is investigated for its role in the oncogenesis of esophageal squamous cell carcinoma (ESCC). CircRNAs are discussed as non-coding RNAs with regulatory potential via miRNA interactions through back-splicing. Using high-throughput RNA sequencing, expression patterns are validated and hsa_circ_0032746 is silenced to assess proliferation, migration, and invasion in vitro and in vivo. Bioinformatic predictions and dual luciferase assays support circRNA/miRNA/mRNA interactions along a miR-4270/MCM3 axis, linking knockdown effects to ESCC progression.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages14

Reading Time35 min

AccessFree to download

Introduction

Background on ESCC and circular RNAs

Materials and methods

Tissue selection and RNA sequencing

Functional assays and silencing strategies

Results

Expression validation of hsa_circ_0032746

CircRNA-miRNA binding and luciferase validation

miR-4270/MCM3 axis and rescue experiments

Conclusion

Shrestha etal. Human Genomics (2024) 18:3 [https://doi.org/10.1186/s40246-023-00564-7](https://doi.org/10.1186/s40246-023-00564-7) Human Genomics RESEARCH Open Access A novel upregulated hsa_circ_ 0032746 regulates the oncogenesis of esophageal squamous cell carcinoma by regulating miR-4270/MCM3 axis Sachin Mulmi Shrestha1, Xin Fang1, Hui Ye2, Lihua Ren2, Qinghua Ji2 and Ruihua Shi1,2* Abstract Introduction Circular RNAs (CircRNA) have emerged as an interest of research in recent years due to its regulatory role in various kinds of cancers of human body. Esophageal squamous cell carcinoma (ESCC) is one of the major disease subtype in Asian countries, including China. CircRNAs are formed by back-splicing covalently joined 3′-and 5′-ends rather than canonical splicing and are found to have binding affinity with miRNAs that conjointly contribute to oncogenesis. Materials and methods 4 pairs of normal, cancer adjacent tissues and cancer tissues were analyzed by highthroughput RNA sequencing and 84 differentially upregulated circRNAs were detected in cancer tissues. hsa_circ_0032746 was silenced by siRNA and lentivirus and then further proliferation, migration and invasion were performed by CCK-8 and transwell assays. Bioinformatic analysis predicted binding affinity of circRNA/miRNA/mRNA axis. Results After qPCR validation, we selected a novel upregulated hsa_circ_0032746 to explore its biogenetic functions which showed high expression in cancer tissues but not in cancer adjacent tissues. The clinicopathological relation of hsa_circ_0032746 showed positive correlation with the tumor location (P = 0 . 026) and gender (P = 0 . 05) . We also predicted that hsa_circ_0032746 could sponge with microRNA. Bioinformatic analysis predicted 11 microRNA response element (MRE) sequences of hsa_circ_0032746 and dual luciferase reporter assay confirmed binding affinity with miR4270 evidencing further study of circRNA/miRNA role. The knockdown of hsa_circ_ 0032746 by siRNA and lentivirus demonstrated that proliferation, invasion and migration of ESCC were inhibited in vitro and vivo experiments. Bioinformatic analysis further predicted MCM3 as a target of miR-4270 and was found upregulated in ESCC upon validation. miR4270 mimic decreased the level of hsa_circ_0032746 and MCM3 while further rescue experiments demonstrated that hsa_circ_0032746 was dependent on miR4270/MCM3 axis on the development process of ESCC. Conclusion We revealed for the first time that circ_ 0032746/mir4270/MCM3 contributes in proliferation, migration and invasion of ESCC and could have potential prognostic and therapeutic significance. Keywords circRNA, Esophageal squamous cell carcinoma, microRNA, Upregulated *Correspondence: Ruihua Shi [ruihuashi@126.com](ruihuashi@126.com) Full list of author information is available at the end of the article © The Author(s) 2024. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creativecommons.org/licenses/by/4.0/](http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ()[. The Creative Commons Public Domain Dedication waiver (](http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication

FAQs

What role does hsa_circ_0032746 play in ESCC?

hsa_circ_0032746 shows high expression in cancer tissues and its knockdown suppresses ESCC proliferation, migration, and invasion, indicating a pro-oncogenic function.

How is hsa_circ_0032746 linked to miR-4270?

Bioinformatics predicts microRNA response elements on hsa_circ_0032746, and the dual luciferase reporter assay confirms binding affinity with miR-4270.

What downstream target is involved in the miR-4270/MCM3 axis?

Bioinformatic analysis predicts MCM3 as a target of miR-4270, and MCM3 is upregulated in ESCC upon validation.

What do the rescue experiments show about the mechanism?

miR-4270 mimic decreases hsa_circ_0032746 and MCM3 levels, and rescue experiments demonstrate that hsa_circ_0032746 depends on the miR-4270/MCM3 axis during ESCC development.

Sep 21, 2026

Uploaded by Jiven