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Molecular Diagnostics

DRD4 promotes chemo-resistance and cancer stem cell-like phenotypes by mediating the activation of the Akt/β-catenin signaling axis in liver cancer

British Journal of Cancer (2024)Cite this article

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Abstract

Background

Liver cancer stem cells (LCSCs) significantly impact chemo-resistance and recurrence in liver cancer. Dopamine receptor D4 (DRD4) is known to enhance the cancer stem cell (CSC) phenotype in glioblastoma and correlates with poor prognosis in some non-central nervous system tumors; however, its influence on LCSCs remains uncertain.

Methods

To investigate the gene and protein expression profiles of DRD4 in LCSCs and non-LCSCs, we utilized transcriptome sequencing and Western blotting analysis. Bioinformatics analysis and immunohistochemistry were employed to assess the correlation between DRD4 expression levels and the pathological characteristics of liver cancer patients. The impact of DRD4 on LCSC phenotypes and signaling pathways were explored using pharmacological or gene-editing techniques. Additionally, the effect of DRD4 on the protein expression and intracellular localization of β-catenin were examined using Western blotting and immunofluorescence.

Results

DRD4 expression is significantly elevated in LCSCs and correlates with short survival in liver cancer. The expression and activity of DRD4 are positive to resistance, self renewal and tumorigenicity in HCC. Mechanistically, DRD4 stabilizes β-catenin and promotes its entry into the nucleus via activating the PI3K/Akt/GSK-3β pathway, thereby enhancing LCSC phenotypes.

Conclusions

Inhibiting DRD4 expression and activation offers a promising targeted therapy for eradicating LCSCs and relieve chemo-resistance.

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Fig. 1: DRD4 expression was significantly increased in HepG2-R and HuH-7-R cells.
Fig. 2: DRD4 is highly expressed in liver cancer specimens and associated with poor prognosis.
Fig. 3: DRD4 activation and overexpression promote multidrug resistance and self-renewal in hepatocellular carcinoma cells.
Fig. 4: DRD4 knockout or inhibition reverses drug resistance and CSC-like phenotypes in HCC cells.
Fig. 5: DRD4 enhances β-catenin expression in the nucleus via PI3K/Akt signaling pathway.
Fig. 6: DRD4 upregulates p-β-catenin(Ser552) and p-GSK3β(Ser9) expression to promote the nucleation of β-catenin by activating PI3K/Akt.
Fig. 7: DRD4, p-Akt, β-catenin, and P-gp are coexpressed in human liver cancer tissues.

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Data availability

Almost all data generated or analysed during this study are included in this manuscript and additional files, while others are available from the corresponding author on reasonable request, except for the information related to patient privacy. RNA-Seq data generated in this study are available on Gene Expression Omnibus (GEO) with accession number GSE272591 at: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE272591.

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Acknowledgements

We sincerely appreciate the contributions from the TCGA, GEO, GSEA, and HPA databases.

Funding

This work was supported by the Youth Fund of National Natural Science Foundation of China (Nos. 81803575 and 31902287), Key R&D and Promotion Projects of Henan Province (No.242102310467), Key Specialized Research and Promotion Project of Henan Province in 2023 (No. 232102311205), Henan Medical Science and Technology Research Program Project (No. LHGJ20210801), Program for Innovative Talents of Science and Technology in Henan Province (No. 23HASTIT043), College Students Innovation and Entrepreneurship Training Program of Henan University (No. 20231022007).

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Author notes

  1. These authors contributed equally: Zhengyan Yang, Pai Zhang, Yiwei Zhao.

Authors and Affiliations

  1. Henan International Joint Laboratory for Nuclear Protein Regulation, Department of Pathology, School of Basic Medical Sciences, Henan University, Kaifeng, 475004, China

    Zhengyan Yang, Yiwei Zhao, Ran Guo, Jinglin Hu, Qi Wang, Handi Liu, Shuangyu Lv & Zhiguang Ren

  2. Medical and Industrial Interdisciplinary Research Institute, Henan University, Kaifeng, 475004, China

    Zhengyan Yang, Qi Wang, Shuangyu Lv, Zhiguang Ren & Daxiang Cui

  3. Joint National Laboratory for Antibody Drug Engineering, Henan University, Kaifeng, 475004, China

    Pai Zhang, Yiwei Zhao & Yanzhong Hu

  4. Genetics and Prenatal Diagnosis Department, Luoyang Maternal and Child Health Hospital, Luoyang, 471023, China

    Pai Zhang

  5. Henan University-affiliated Zhengzhou Yihe Hospital, Zhengzhou, 450000, China

    Zhi Zhao

  6. Key Laboratory of Clinical Resources Translation, The First Affiliated Hospital, Henan University, Kaifeng, 475004, China

    Zhiguang Ren

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  1. Zhengyan Yang
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Contributions

ZY, PZ, YZ, RG, JH, QW, and HL participated in the experimental data collection; ZZ provided technical assistance; ZR designed experiments and drafted the manuscript; SL, YH, and DC helped revise the manuscript. All authors contributed to the article and approved the submitted version.

Corresponding authors

Correspondence to Zhiguang Ren, Yanzhong Hu or Daxiang Cui.

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Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

The experiments involving animal studies and patient specimens were reviewed and approved by the Ethics Committee of the Medical School of Henan University (HUSOM2023-221 and HUSOM2023-222). Informed consent was obtained from all participants involved in the study. All authors agree with the content of the manuscript.

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Yang, Z., Zhang, P., Zhao, Y. et al. DRD4 promotes chemo-resistance and cancer stem cell-like phenotypes by mediating the activation of the Akt/β-catenin signaling axis in liver cancer. Br J Cancer (2024). https://doi.org/10.1038/s41416-024-02811-7

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