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MiR-330-5p and miR-1270 target essential components of RNA polymerase I transcription and exhibit a novel tumor suppressor role in lung adenocarcinoma

Cancer Gene Therapy volume 30, pages 288–301 (2023)Cite this article

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Abstract

Upregulation of RNA polymerase I (Pol I) transcription and the overexpression of Pol I transcriptional machinery are crucial molecular alterations favoring malignant transformation. However, the causal molecular mechanism(s) of this aberration remain largely unknown. Here, we found that Pol I transcription and its core machinery are upregulated in lung adenocarcinoma (LUAD). We show that the loss of miRNAs (miR)-330-5p and miR-1270 expression contributes to the upregulation of Pol I transcription in LUAD. Constitutive overexpression of these miRs in LUAD cell lines suppressed the expression of core components of Pol I transcription, and reduced global ribosomal RNA synthesis. Importantly, miR-330-5p/miR-1270-mediated repression of Pol I transcription exerted multiple tumor suppressive functions including reduced proliferation, cell cycle arrest, enhanced apoptosis, reduced migration, increased drug sensitivity, and reduced tumor burden in a mouse xenograft model. Mechanistically, the downregulation of miR-330-5p and miR-1270 is regulated by Pol I subunit-derived circular RNA circ_0055467 and DNA hypermethylation, respectively. This study uncovers a novel miR-330-5p/miR-1270 mediated post-transcriptional regulation of Pol I transcription, and establish tumor suppressor properties of these miRs in LUAD. Ultimately, our findings provide a rationale for the therapeutic targeting of Pol I transcriptional machinery for LUAD.

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Fig. 1: Pol I transcription in LUAD.
Fig. 2: miR-330-5p and miR-1270 target PIC components of Pol I transcription.
Fig. 3: Overexpression of miR-330-5p and miR-1270 inhibits rRNA synthesis.
Fig. 4: Effect of miR-330-5p and miR-1270 on cell proliferation, cell cycle and apoptosis.
Fig. 5: Effect of miR-330-5p and miR-1270 on cell migration and cisplatin sensitivity.
Fig. 6: Regulation of miR-330-5p and miR-1270.
Fig. 7: MiR-330-5p and miR-1270 reduce tumor burden in-vivo.
Fig. 8: Graphical summary depicting regulatory networks involved in the regulation of Pol I transcription in LUAD.

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Data availability

The data generated in this study are available within the article and its supplementary data files. TCGA gene expression data for LUAD is available from UCSC Xena browser. The CpG island methylation track (cg19204924) of LUAD or normal is available from TCGA Wanderer database (http://maplab.imppc.org/wanderer/).

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Acknowledgements

We are grateful to IIT Ropar for funding. We thank the animal facility at IISER, Mohali, Punjab, India for in-vivo studies. We thank Dr. Juhi Tayal and Dr. Anurag Mehta, Biorepository facility, RGCIRC, New Delhi, India, for providing tumor samples. We heartfully thank Prof. Galande IISER Pune, for H23 cells and Dr. Harshan, CCMB Hyderabad, for ZEB1 expression vector. Special thanks to Arpita Karmakar and Deepika for their help during revision work, and Sheetal Yadav for manuscript editing.

Funding

Seed grant—IIT Ropar granted to Dr. Srivatsava Naidu.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Indian Institute of Technology Ropar, Rupnagar, Punjab, India

    Sheetanshu Saproo, Shashanka S. Sarkar, Ekta Gupta, Sourav Chattopadhyay, Aarzoo Charaya & Srivatsava Naidu

  2. Department of Computational Biology, Indraprastha Institute of Information Technology-Delhi, New Delhi, India

    Siddhant Kalra & Gaurav Ahuja

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  1. Sheetanshu Saproo
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Contributions

S.N. conceived and designed the study, interpreted the data, drafted and revised the manuscript. S.S. acquired data, handled animal work and played role in interpreting the results, drafted the manuscript. E.G. acquired data and prepared figures. S.S.S. acquired data, played a role in computational analysis. S.C. and A.C. contributed in animal work and project management. S.K., G.A. contributed in computational analysis. All authors approved the final version, agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

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Correspondence to Srivatsava Naidu.

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The authors declare no competing interests.

Ethical approval

Mice work protocols were approved (IISERM/SAFE/PRT/2020/005) by the institutional animal ethics committee, Indian Institute of Science Education and Research (IISER) Mohali, as per the standards of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), and comply with the ARRIVE guidelines. Frozen specimens of LUAD or NAT were obtained from Rajiv Gandhi Cancer Institute and Research Centre (RGCI&RC), New Delhi, India, in accordance with the guidelines of the institutional review board (Approval number: RGCIRC/IRB/343/2019) and the Declaration of Helsinki.

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Saproo, S., Sarkar, S.S., Gupta, E. et al. MiR-330-5p and miR-1270 target essential components of RNA polymerase I transcription and exhibit a novel tumor suppressor role in lung adenocarcinoma. Cancer Gene Ther 30, 288–301 (2023). https://doi.org/10.1038/s41417-022-00544-4

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