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Identification of circATG9A as a novel biomarker for renal cell carcinoma

Cancer Gene Therapy volume 31, pages 82–93 (2024)Cite this article

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Abstract

The incidence and mortality rates of renal cell carcinoma (RCC) have rapidly increased worldwide. To gain new insights into the regulatory role of circular RNAs (circRNAs) in RCC progression, we conducted RNA sequencing on three pairs of ccRCC and adjacent normal tissues. RT-PCR was utilized to analyze RNA expression. We investigated the effects of circATG9A on RCC cells through various assays including CCK-8, Transwell, wound healing, and colony formation assays. Furthermore, we employed FISH, RNA pull-down, luciferase reporter, and RIP assays to elucidate the mechanism by which circATG9A regulates RCC. Ultimately, we identified 118 differentially expressed circRNAs in RCC, including a novel circRNA, circATG9A, which was found to promote RCC progression both in vitro and in vivo. Moreover, mRNA sequencing, western blotting, and rescue experiments indicated that TRPM3 is the target of circATG9A in RCC progression. Bioinformatic analysis, RNA pull-down, FISH, and RIP assays suggested that circATG9A regulates TRPM3 expression by acting as a sponge for miR-497-5p. Finally, Western blotting revealed that circATG9A promotes the epithelial-mesenchymal transition (EMT) process through the Wnt/β-catenin signaling pathway. Our findings demonstrate that circATG9A is a novel circRNA upregulated in RCC that plays a crucial role in the EMT process through the miR-497-5p/TRPM3/Wnt/β-catenin axis. These results suggest that circATG9A could be a promising target for RCC prognosis and therapy.

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Fig. 1: The identification of the DE circRNAs in RCC (Drawn by BioRender).
Fig. 2: The characteristics of circATG9A.
Fig. 3: CircATG9A facilitated the progression of RCC cells in vivo.
Fig. 4: TRPM3 is the target of circATG9A in RCC progression.
Fig. 5: CircATG9A regulates the expression of TRMP3 by sponging miR-497-5p.
Fig. 6: MiR-497-5p could rescue the facilitative effect of circATG9A on the phenotypes of CCR cells.
Fig. 7: CircATG9A promotes the epithelial–mesenchymal transition (EMT) process via WNT/β‐catenin signaling pathway in RCC.
Fig. 8: CircATG9A could promote RCC progression and EMT in vivo.
Fig. 9: The schematic diagram of CircATG9A facilitating the process RCC is drawn using BioRender.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Author notes

  1. These authors contributed equally: Ye Shen, Ruipeng Wu

Authors and Affiliations

  1. Department of Urology, Northern Jiangsu People’s Hospital, Clinical Medical College of Yangzhou University, Yangzhou, China

    Ye Shen, Ruipeng Wu, Zou Zhuo, Ximeng Deng, Weijian Li & Changkun Liu

  2. Graduate School, Dalian Medical University, Dalian, China

    Ruipeng Wu & Zou Zhuo

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  1. Ye Shen
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Contributions

CL conceived and designed this experiment. YS and WL performed the experiments and collected the samples. RW and XD performed the data analysis. YS and ZZ wrote the manuscript. CL performed manuscript revision. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Ye Shen or Changkun Liu.

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The authors declare no competing interests.

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The animal studies got the approval of the Ethics Committee of Yangzhou University (NO. 202212002).

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The collection of tissue samples was approved by the Ethics Committee of the Yangzhou University. Written informed consent was obtained from the patients before collecting all tissues.

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Shen, Y., Wu, R., Zhuo, Z. et al. Identification of circATG9A as a novel biomarker for renal cell carcinoma. Cancer Gene Ther 31, 82–93 (2024). https://doi.org/10.1038/s41417-023-00684-1

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