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

---

M. E. El-Houseini¹, A. Ismail², A. A. Abdelaal², A. H. El-Habashy³, Z. F. Abdallah¹, M. Z. Mohamed⁴, M. El-Hadidi⁵, W. C. S. Cho⁶, H. Ahmed⁷, and T. A. Al-Shafie^1,8,a,b,c*

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

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

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

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

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

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

⁷GlycoMantra, Inc., Baltimore MD 21227, USA

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

^* To whom correspondence should be addressed.

Received March 19, 2019; Revised April 19, 2019; Accepted April 20, 2019

---

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.

---

KEY WORDS: hepatocellular carcinoma, chronic liver disease, TGF-β1, liver stem cells, C-Kit mutations

DOI: 10.1134/S0006297919080108

---