Abstract
Background
Autophagy plays multifaceted roles in regulating hepatocellular carcinoma (HCC) and the mechanisms involved are under-explored. Regulatory microRNAs (miRNAs) have been reported to target autophagy proteins but their roles in HCC is not well studied. Using HCC patient tissues, this study aims to investigate the association of autophagy with several clinicopathological parameters as well as identifying the autophagy-related miRNAs and the possible pathways.
Methods and results
Autophagy level in the HCC patient-derived cancer and non-cancer tissues was determined by immunohistochemistry (IHC) targeting SQSTM1, LC3A and LC3B proteins. Significance tests of clinicopathological variables were tested using the Fisher’s exact or Chi-square tests. Gene and miRNA expression assays were carried out and analyzed using Nanostring platform and software followed by validation of other online bioinformatics tools, namely String and miRabel. Autophagy expression was significantly higher in cancerous tissues compared to adjacent non-cancer tissues. High LC3B expression was associated with advanced tumor histology grade and tumor location. Nanostring gene expression analysis revealed that SQSTM1, PARP1 and ATG9A genes were upregulated in HCC tissues compared to non-cancer tissues while SIRT1 gene was downregulated. These genes are closely related to an autophagy pathway in HCC. Further, using miRabel tool, three downregulated miRNAs (hsa-miR-16b-5p, hsa-miR-34a-5p, and hsa-miR-660-5p) and one upregulated miRNA (hsa-miR-539-5p) were found to closely interact with the abovementioned autophagy-related genes. We then mapped out the possible pathway involving the genes and miRNAs in HCC tissues.
Conclusions
We conclude that autophagy events are more active in HCC tissues compared to the adjacent non-cancer tissues. We also reported the possible role of several miRNAs in regulating autophagy-related genes in the autophagy pathway in HCC. This may contribute to the development of potential therapeutic targets for improving HCC therapy. Future investigations are warranted to validate the target genes reported in this study using a larger sample size and more targeted molecular technique.
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Data availability
No datasets were generated or analysed during the current study.
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Acknowledgements
Magdelyn Wong is the recipient of Sunway University Postgraduate’s Degree by Research Studentship. We also thank Wenzhou-Kean University for continuously supporting the research work.
Funding
This work was partly funded by Ministry of Higher Education Malaysia (FRGS/1/2019/SKK08/SYUC/02/1) and Sunway Medical Centre Research Funds (SRC/002/2017/FR and SRC/003/ 2017/ FR). We also thank Wenzhou-Kean University (WKU) Student Partnering with Faculty (SpF) Research Program [grant number WKUSPF2023025 and WKUSPF2023026], Internal Start-Up Research Grant [grant number ISRG2023030] and Zhejiang Province Department of Education General Program 2023 [grant number Y202353358] for partly supporting this work.
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All authors contributed to the study conception and design as well as the analysis and interpretation of the data. Material preparation and data collection were performed by Magdelyn Mei-Theng Wong. The first draft of the manuscript was written by Magdelyn Mei-Theng Wong and Sin-Yeang Teow. Sin-Yeang Teow reviewed and edited the manuscript. All authors read and approved the final manuscript.
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This study was performed in line with the principles of the Declaration of Helsinki. Ethical approval for this study was granted from Sunway University Research Ethics Committee (SUNREC 2017/051) and Sunway Medical Centre Independent Research Ethics Committee (013/2017/ER). Archived FFPE tissue blocks of HCC patients from year 2012 to 2016 from Sunway Medical Centre were used in this project. The written informed consent was waived with the approval of the Institutional Review Board (IRB).
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Wong, M.MT., Aziz, N.A., Ch’ng, E.S. et al. Expression of LC3A, LC3B and p62/SQSTM1 autophagy proteins in hepatocellular carcinoma (HCC) tissues and the predicted microRNAs involved in the autophagy-related pathway. J Mol Histol (2024). https://doi.org/10.1007/s10735-024-10191-8
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DOI: https://doi.org/10.1007/s10735-024-10191-8