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Investigating the roles of microRNAs associated with cancer stem cells, drug resistance, metastasis and recurrence in hepatocellular carcinoma: A systematic review, network analysis and experimental verification

  • Authors:
    • Yuanyuan Jiang
    • Xiaoning Zhu
    • Qinghua Peng
    • Paniz Zarghami Dastjerdi
    • Mengyun Peng
    • Ding Zheng
    • Xiaodong Wang
    • Yoke Kqueen Cheah
    • Jing Wang
  • View Affiliations / Copyright

    Affiliations: Hepatobiliary Department, The Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University, Luzhou, Sichuan 646000, P.R. China, Department of Biomedical Science, Institute of Bioscience, University Putra Malaysia, Serdang, Selangor 43400, Malaysia, Department of Biomedical Science, Faculty of Medicine and Health Sciences, University Putra Malaysia, Serdang, Selangor 43400, Malaysia
    Copyright: © Jiang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 3
    |
    Published online on: October 23, 2025
       https://doi.org/10.3892/ol.2025.15356
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Abstract

Cancer stem cells (CSCs) are responsible for drug therapy resistance, recurrence and metastasis in hepatocellular carcinoma (HCC). Certain microRNAs (miRNAs/miRs) are involved in various pathological cancer pathways via their binding interactions with target mRNAs. The objectives of the present study were to explore the potential microRNAs associated with HCC‑CSCs, and to investigate their roles in recurrence, metastasis and drug resistance. Initially, a registered systematic review (CRD42024508526; International Prospective Register of Systematic Reviews) identified a group of miRNAs associated with HCC prognosis, and the Coremine Medical data mining tool identified another group of potential miRNAs associated with HCC‑CSCs, recurrence, metastasis and drug resistance. Secondly, potential miRNAs that were associated with HCC prognosis, CSCs, recurrence, metastasis and drug resistance were detected by comparing the two groups of miRNAs. Subsequently, the expression levels of a potential miRNA in HCC tissues and its prognostic predictive power were evaluated using the Encyclopedia of RNA Interactomes and Kaplan‑Meier plotter online tools. In addition, the expression levels of the target miRNA in Huh7‑CSCs and Huh7 cells were measured using reverse transcription‑quantitative PCR (RT‑qPCR). Finally, the potential biological processes of the miRNA target genes were examined through bioinformatics analysis using the Enrichr database. Briefly, hsa‑miR‑17‑5p was predicted to be associated with stemness, metastasis, recurrence and drug resistance in HCC. Bioinformatics analysis demonstrated that miR‑17‑5p expression was higher in HCC tissues compared with that in para‑tumor tissues and that patients with low miR‑17‑5p expression demonstrated higher overall survival rates (months). The RT‑qPCR results indicated that miR‑17‑5p expression in Huh7‑CSCs was significantly higher compared with that in Huh7 cells. Further bioinformatics analysis suggested that miR‑17‑5p maintains stemness by targeting hypoxia‑inducible factor‑1α (HIF1A) and Myc. Additionally, the target genes of miR‑17‑5p were revealed to be involved in cell fate and metabolic reprogramming pathways. In conclusion, miR‑17‑5p may be a potential miRNA associated with CSCs, metastasis, recurrence and drug resistance in HCC via cell fate and metabolic reprogramming pathways. miR‑17‑5p exhibited higher expression in HCC‑CSCs compared with that in HCC cell lines, and may target HIF1A and Myc to maintain HCC‑CSCs stemness.
View Figures

Figure 1

Flow diagram depicting the systematic
review article selection process.

Figure 2

miR-17-5p is a miRNA potentially
associated with CSCs, metastasis, recurrence and drug resistance in
HCC. (A) Venn diagram illustrating how the potential miRNA
(miR-17-5p) was selected by exploring the intersection between the
nine miRNAs associated with the prognosis of HCC, and the 34 miRNAs
associated with HCC-CSCs, metastasis, tumor recurrence and drug
resistance. (B) Box plot of miR-17-5p expression in LIHC. (C)
Survival analysis of miR-17-5p in LIHC. (D) Interaction network of
targeted genes of miR-17-5p, with the 12 genes involved in
metastasis, stemness and drug resistance specially marked.
miR/miRNA, microRNA; CSC, cancer stem cell; HCC, hepatocellular
carcinoma; LIHC, liver hepatocellular carcinoma; RPM, reads per
million.

Figure 3

Morphology of HCC-CSCs and expression
levels of miR-17-5p. (A) Huh7 cell line and Huh7-CSCs
(magnification, ×10). (B) Expression levels of miR-17-5p in Huh7
and Huh7-CSCs. **P<0.005. CSC, cancer stem cell; HCC,
hepatocellular carcinoma; miR, microRNA.
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Jiang Y, Zhu X, Peng Q, Dastjerdi PZ, Peng M, Zheng D, Wang X, Cheah YK and Wang J: Investigating the roles of microRNAs associated with cancer stem cells, drug resistance, metastasis and recurrence in hepatocellular carcinoma: A systematic review, network analysis and experimental verification. Oncol Lett 31: 3, 2026.
APA
Jiang, Y., Zhu, X., Peng, Q., Dastjerdi, P.Z., Peng, M., Zheng, D. ... Wang, J. (2026). Investigating the roles of microRNAs associated with cancer stem cells, drug resistance, metastasis and recurrence in hepatocellular carcinoma: A systematic review, network analysis and experimental verification. Oncology Letters, 31, 3. https://doi.org/10.3892/ol.2025.15356
MLA
Jiang, Y., Zhu, X., Peng, Q., Dastjerdi, P. Z., Peng, M., Zheng, D., Wang, X., Cheah, Y. K., Wang, J."Investigating the roles of microRNAs associated with cancer stem cells, drug resistance, metastasis and recurrence in hepatocellular carcinoma: A systematic review, network analysis and experimental verification". Oncology Letters 31.1 (2026): 3.
Chicago
Jiang, Y., Zhu, X., Peng, Q., Dastjerdi, P. Z., Peng, M., Zheng, D., Wang, X., Cheah, Y. K., Wang, J."Investigating the roles of microRNAs associated with cancer stem cells, drug resistance, metastasis and recurrence in hepatocellular carcinoma: A systematic review, network analysis and experimental verification". Oncology Letters 31, no. 1 (2026): 3. https://doi.org/10.3892/ol.2025.15356
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Spandidos Publications style
Jiang Y, Zhu X, Peng Q, Dastjerdi PZ, Peng M, Zheng D, Wang X, Cheah YK and Wang J: Investigating the roles of microRNAs associated with cancer stem cells, drug resistance, metastasis and recurrence in hepatocellular carcinoma: A systematic review, network analysis and experimental verification. Oncol Lett 31: 3, 2026.
APA
Jiang, Y., Zhu, X., Peng, Q., Dastjerdi, P.Z., Peng, M., Zheng, D. ... Wang, J. (2026). Investigating the roles of microRNAs associated with cancer stem cells, drug resistance, metastasis and recurrence in hepatocellular carcinoma: A systematic review, network analysis and experimental verification. Oncology Letters, 31, 3. https://doi.org/10.3892/ol.2025.15356
MLA
Jiang, Y., Zhu, X., Peng, Q., Dastjerdi, P. Z., Peng, M., Zheng, D., Wang, X., Cheah, Y. K., Wang, J."Investigating the roles of microRNAs associated with cancer stem cells, drug resistance, metastasis and recurrence in hepatocellular carcinoma: A systematic review, network analysis and experimental verification". Oncology Letters 31.1 (2026): 3.
Chicago
Jiang, Y., Zhu, X., Peng, Q., Dastjerdi, P. Z., Peng, M., Zheng, D., Wang, X., Cheah, Y. K., Wang, J."Investigating the roles of microRNAs associated with cancer stem cells, drug resistance, metastasis and recurrence in hepatocellular carcinoma: A systematic review, network analysis and experimental verification". Oncology Letters 31, no. 1 (2026): 3. https://doi.org/10.3892/ol.2025.15356
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