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Role of TGF-β1 and C-Kit Mutations in the Development of Hepatocellular Carcinoma in Hepatitis C Virus-Infected Patients: in vitro Study

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Abstract

Transforming growth factor beta (TGF-β) acts as a tumor-suppressing cytokine in healthy tissues and non-malignant tumors. Yet, in malignancy, TGF-β can exert the opposite effects that can promote proliferation of cancer cells. C-Kit plays a prominent role in stem cell activation and liver regeneration after injury. However, little is known about the cross-talk between TGF-β and C-Kit and its role in the progression of hepatocellular carcinoma (HCC). Here, we studied the effect of increasing doses of TGF-β1 on CD44+CD90+ liver stem cells (LSCs) and C-Kit gene expression in malignant and adjacent non-malignant liver tissues excised from 32 HCC patients. The percentage of LSCs in malignant tumors was two times higher compared to their counterparts from the non-malignant tissues. When treated with increasing doses of TGF-β1, proliferation of both malignant and non-malignant LSCs was progressively suppressed, but low TGF-β1 dose failed to suppress the growth of malignant LSCs. Moreover, C-Kit exons 9 and 11 were expressed in malignant LSCs, but not in their non-malignant counterparts. Analysis of C-Kit detected mutations in exon 9 (but not in exon 11) in some malignant liver cells resulting in the changes in the amino acid sequence and dysregulation of protein structure and function. Interestingly, in malignant liver cells, mutations in exon 9 were associated with high-viremia hepatitis C virus (HCV), and expression of this exon was not suppressed by the TGF-β1 treatment at all doses. To our knowledge, this is the first report that mutations in the C-Kit gene in HCC patients are associated with high-viremia HCV. Our study emphasizes the need for investigation of the TGF-β1 level and C-Kit mutations in patients with chronic HCV for HCC prevention and better therapy management.

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Abbreviations

C-Kit:

stem cell factor receptor

CSC:

cancer stem cell

HCC:

hepatocellular carcinoma

HCV:

hepatitis C virus

LSC:

liver stem cell

Peg-INF:

pegylated interferon

SCF:

stem cell factor

TGF-β1:

transforming growth factor beta 1

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Acknowledgments

This study was supported by the National Cancer Institute, Cairo University, Egypt. The authors would like to thank all members of the Cancer Biology Department for their help.

Funding

Funding. This study was supported in part by the National Institutes of Health (grant CA203420 to HA).

Author information

Authors and Affiliations

  1. Cairo University, National Cancer Institute, Department of Cancer Biology, 11796, Cairo, Egypt

    M. E. El-Houseini, Z. F. Abdallah & T. A. Al-Shafie

  2. Ain Shams University, Faculty of Medicine, Department of Surgery, 11566, Cairo, Egypt

    A. Ismail & A. A. Abdelaal

  3. Cairo University, National Cancer Institute, Department of Pathology, 11796, Cairo, Egypt

    A. H. El-Habashy

  4. Medical Center of Egyptian Railways, Department of Medical Laboratory, 11669, Cairo, Egypt

    M. Z. Mohamed

  5. Nile University, Center of Informatics Science, 12525, Giza, Egypt

    M. El-Hadidi

  6. Queen Elizabeth Hospital, Department of Clinical Oncology, Kowloon, Hong Kong, China

    W. C. S. Cho

  7. GlycoMantra, Inc., Baltimore, MD, 21227, USA

    H. Ahmed

  8. Pharos University in Alexandria, Faculty of Pharmacy and Drug Manufacturing, Department of Pharmacology and Therapeutics, 21311, Alexandria, Egypt

    T. A. Al-Shafie

Authors
  1. M. E. El-Houseini
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  2. A. Ismail
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  3. A. A. Abdelaal
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  4. A. H. El-Habashy
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  5. Z. F. Abdallah
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  6. M. Z. Mohamed
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  7. M. El-Hadidi
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  8. W. C. S. Cho
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  9. H. Ahmed
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  10. T. A. Al-Shafie
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Contributions

Authors’ contributions. All authors were involved in the preparation of this manuscript. MZ collected the data; TA, HA, and WC wrote the manuscript. AI and AA performed surgery and designed the study. MH, ZF, and MH summarized the data and revised the manuscript. AH substantially contributed to the study design, performed surgery, and revised the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to T. A. Al-Shafie.

Ethics declarations

Conflict of interest. The authors declare no conflict of interest.

Ethical approval. The study was carried out with permission from the Institution Review Board (IRB) of National Cancer Institute, Cairo University (#IRB00004025) in accordance with applicable institutional and international regulations and guidelines and confirmed to the provisions of the Declaration of Helsinki.

Patient consent for publication. All patients were informed of the investigational nature of this study and provided their written informed consent.

Availability of data and materials. The datasets generated and/or analyzed in this study are available from http://www.chemcomp.com.

Additional information

Published in Russian in Biokhimiya, 2019, Vol. 84, No. 8, pp. 1189–1204.

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El-Houseini, M.E., Ismail, A., Abdelaal, A.A. et al. Role of TGF-β1 and C-Kit Mutations in the Development of Hepatocellular Carcinoma in Hepatitis C Virus-Infected Patients: in vitro Study. Biochemistry Moscow 84, 941–953 (2019). https://doi.org/10.1134/S0006297919080108

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