Identification of the possible therapeutic targets in the insulin-like growth factor 1 receptor pathway in a cohort of Egyptian hepatocellular carcinoma complicating chronic hepatitis C type 4
Background: Molecular targeted drugs are the first line of treatment of advanced hepatocellular carcinoma (HCC) due to its chemo- and radioresistant nature. HCC has several well-documented etiologic factors that drive hepatocarcinogenesis through different molecular pathways. Currently, hepatitis C virus (HCV) is a leading cause of HCC. Therefore, we included a unified cohort of HCV genotype 4-related HCCs to study the expression levels of genes involved in the insulin-like growth factor 1 receptor (IGF1R) pathway, which is known to be involved in all aspects of cancer growth and progression.
Aim: Determine the gene expression patterns of IGF1R pathway genes in a cohort of Egyptian HCV-related HCCs. Correlate them with different patient/tumor characteristics. Determine the activity status of involved pathways.
Methods: Total ribonucleic acid (RNA) was extracted from 32 formalin-fixed paraffin-embedded tissues of human HCV-related HCCs and 6 healthy liver donors as controls. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) using RT2 Profiler PCR Array for Human Insulin Signaling Pathway was done to determine significantly up- and downregulated genes with identification of most frequently coregulated genes, followed by correlation of gene expression with different patient/tumor characteristics. Finally, canonical pathway analysis was performed using the Ingenuity Pathway Analysis software.
Results: Six genes – AEBP1, AKT2, C-FOS, PIK3R1, PRKCI, SHC1 – were significantly overexpressed. Thirteen genes – ADRB3, CEBPA, DUSP14, ERCC1, FRS3, IGF2, INS, IRS1, JUN, MTOR, PIK3R2, PPP1CA, RPS6KA1 – were significantly underexpressed. Several differentially expressed genes were related to different tumor/patient characteristics. Nitric oxide and reactive oxygen species production pathway was significantly activated in the present cohort, while the growth hormone signaling pathway was inactive.
Conclusions: The gene expression patterns identified in this study may serve as possible therapeutic targets in HCV-related HCCs. The most frequently coregulated genes may serve to guide combined molecular targeted therapies. The IGF1R pathway showed evidence of inactivity in the present cohort of HCV-related HCCs, so targeting this pathway in therapy may not be effective.
International Agency for Research on Cancer (IARC). Globocan 2012: Estimated cancer incidence, mortality and prevalence worldwide in 2012. France: IARC; 2012.
Shi J, Zhu L, Liu S, Xie WF. A meta-analysis of case-control studies on the combined effect of hepatitis B and C virus infections in causing hepatocellular carcinoma in China. Br J Cancer. 2005;92(3):607–612.
Llovet JM, Villanueva A, Lachenmayer A, Finn RS. Advances in targeted therapies for hepatocellular carcinoma in the genomic era. Nat Rev Clin Oncol. 2015;12(7):408–424.
El-zanaty F, Way A. Egypt Demographic and Health Survey 2008. Cairo, Egypt: Ministry of Health, El-Zanaty and Associates, and Macro International; 2009.
Maucort-Boulch D, de Martel C, Franceschi S, Plummer M. Fraction and incidence of liver cancer attributable to hepatitis B and C viruses worldwide. Int J Cancer. 2018;142(12):2471-7.
Ray SC, Arthur RR, Carella A, Bukh J, Thomas DL. Genetic epidemiology of hepatitis C virus throughout Egypt. J Infect Dis. 2000;182(3):698–707.
Wang Y, Sun Y. Insulin-like growth factor receptor-1 as an anti-cancer target: blocking transformation and inducing apoptosis. Curr Cancer Drug Targets. 2002;2(3):191–207.
Christopoulos PF, Msaouel P, Koutsilieris M. The role of the insulin-like growth factor-1 system in breast cancer. Mol Cancer. 2015;14(1):43.
Papa V, Gliozzo B, Clark GM, et al. Insulin-like growth factor-I receptors are overexpressed and predict a low risk in human breast cancer. Cancer Res. 1993;53(16):3736–3740.
Jones R, Campbell C, Gunther E, et al. Transgenic overexpression of IGF-IR disrupts mammary ductal morphogenesis and induces tumor formation. Oncogene. 2007;26(11):1636–1644.
Samani AA, Yakar S, LeRoith D, Brodt P. The role of the IGF system in cancer growth and metastasis: overview and recent insights. Endocr Rev. 2007;28(1):20–47.
Vescovo T, Refolo G, Vitagliano G, Fimia GM, Piacentini M. Molecular mechanisms of hepatitis c virus–induced hepatocellular carcinoma. Clin Microbiol Infect. 2016;22(10):853–861.
Patel A, Sun W. Molecular targeted therapy in hepatocellular carcinoma: from biology to clinical practice and future. Curr Treat Opt Oncol. 2014;15(3):380–394.
Lu L-C, Hsu C-H, Hsu C, Cheng A-L. Tumor heterogeneity in hepatocellular carcinoma: facing the challenges. Liver Cancer. 2016;5(2):128–138.
Luo JH, Ren B, Keryanov S, et al. Transcriptomic and genomic analysis of human hepatocellular carcinomas and hepatoblastomas. Hepatology. 2006;44(4):1012–1024.
Lee JS, Chu IS, Heo J, et al. Classification and prediction of survival in hepatocellular carcinoma by gene expression profiling. Hepatology. 2004;40(3):667–676.
Shirota Y, Kaneko S, Honda M, Kawai HF, Kobayashi K. Identification of differentially expressed genes in hepatocellular carcinoma with cDNA microarrays. Hepatology. 2001;33(4):832–840.
Tackels-Horne D, Goodman MD, Williams AJ, et al. Identification of differentially expressed genes in hepatocellular carcinoma and metastatic liver tumors by oligonucleotide expression profiling. Cancer. 2001;92(2):395–405.
Patil MA, Chua M-S, Pan K-H, et al. An integrated data analysis approach to characterize genes highly expressed in hepatocellular carcinoma. Oncogene. 2005;24(23):3737–3747.
Guimei M, Baddour N, ElKaffash D, Abdou L, Taher Y. Gremlin in the pathogenesis of hepatocellular carcinoma complicating chronic hepatitis C: an immunohistochemical and PCR study of human liver biopsies. BMC Res Notes. 2012;5(1):390.
Huynh H. Molecularly targeted therapy in hepatocellular carcinoma. Biochem Pharmacol. 2010;80(5):550–560.
Saez E, Rutberg SE, Mueller E, et al. C-fos is required for malignant progression of skin tumors. Cell. 1995;82(5):721–732.
Yuen MF, Wu PC, Lai VCH, Lau JYN, Lai CL. Expression of c-Myc, c-Fos, and c-Jun in hepatocellular carcinoma. Cancer. 2001; 91(1):106–112.
Kamide D, Yamashita T, Araki K, et al. Selective activator protein-1 inhibitor t-5224 prevents lymph node metastasis in an oral cancer model. Cancer Sci. 2016;107(5):666–673.
Wang L, Yao J, Zhang X, et al. Mirna-302b suppresses human hepatocellular carcinoma by targeting Akt2. Mol Cancer Res. 2014;12(2):190–202.
Zhang Y, Guo X, Yang M, Yu L, Li Z, Lin N. Identification of AKT kinases as unfavorable prognostic factors for hepatocellular carcinoma by a combination of expression profile, interaction network analysis and clinical validation. Mol BioSyst. 2014;10(2):215–222.
Kurokawa Y, Matoba R, Takemasa I, et al. Molecular features of non-B, non-C hepatocellular carcinoma: a PCR-array gene expression profiling study. J Hepatol. 2003;39(6):1004–1012.
Huang C-Y, Huang X-P, Zhu J-Y, et al. miR-128-3p suppresses hepatocellular carcinoma proliferation by regulating PIK3R1 and is correlated with the prognosis of HCC patients. Oncol Rep. 2015;33(6):2889–2898.
Wang JM, Li Q, Du GS, Lu JX, Zou SQ. Significance and expression of atypical protein kinase C-iota in human hepatocellular carcinoma. J Surg Res. 2009;154(1):143–149.
Chen X, Cheung ST, So S, et al. Gene expression patterns in human liver cancers. Mol Biol Cell. 2002;13(6):1929–1939.
Ro H-S, Roncari D. The C/EBP-binding region and adjacent sites regulate expression of the adipose p2 gene in human preadipocytes. Mol Cell Biol. 1991;11(4):2303–2306.
Majdalawieh A, Ro HS. Regulation of IkappaBalpha function and NF-kappaB signaling: AEBP1 is a novel proinflammatory mediator in macrophages. Mediators Inflamm. 2010;2010:823–821.
Zhang L, Reidy SP, Nicholson TE, et al. The role of AEBP1 in sex-specific diet-induced obesity. Mol Med. 2005;11(1–12):39.
Ladha J, Sinha S, Bhat V, Donakonda S, Rao SM. Identification of genomic targets of transcription factor AEBP1 and its role in survival of glioma cells. Mol Cancer Res. 2012;10(8):1039–1051.
Han S-J, Cho YL, Nam GH, Kim CK, Seo J-S, Ahn WS. CDNA microarray analysis of gene expression profiles associated with cervical cancer. Cancer Res Treat. 2003;35(5):451–459.
Ahn WS, Bae SM, Lee JM, et al. Searching for pathogenic gene functions to cervical cancer. Gynecol Oncol. 2004;93(1):41–48.
Reddy SP, Britto R, Vinnakota K, et al. Novel glioblastoma markers with diagnostic and prognostic value identified through transcriptome analysis. Clin Cancer Res. 2008;14(10):2978–2987.
Grigoriadis A, Mackay A, Reis-Filho JS, et al. Establishment of the epithelial-specific transcriptome of normal and malignant human breast cells based on MPSS and array expression data. Breast Cancer Res. 2006;8(5):R56.
Wang L, Yao J, Sun H, et al. miR-302b suppresses cell invasion and metastasis by directly targeting AKT2 in human hepatocellular carcinoma cells. Tumor Biol. 2016;37(1):847–855.
Jilkova ZM, Kuyucu AZ, Kurma K, et al. Combination of AKT inhibitor ARQ 092 and sorafenib potentiates inhibition of tumor progression in cirrhotic rat model of hepatocellular carcinoma. Oncotarget. 2018;9(13):11145–11158.
Ma C, Yang Y, Wang J, et al. The aPKCι blocking agent atm negatively regulates EMT and invasion of hepatocellular carcinoma. Cell Death Dis. 2014;5(3):e1129.
Acevedo-Duncan M. Method of treating colorectal cancers using a PKC inhibitor. In: Google Patents; 2018.
Wang J-M, Li Q, Du G-S, Lu J-X, Zou S-Q. Significance and expression of atypical protein kinase c-ι in human hepatocellular carcinoma. J Surg Res. 2009;154(1):143–149.
Du G-S, Wang J-M, Lu J-X, et al. Expression of P-aPKC-ι, E-cadherin, and β-catenin related to invasion and metastasis in hepatocellular carcinoma. Ann Surg Oncol. 2009;16(6):1578–1586.
Chiang DY, Villanueva A, Hoshida Y, et al. Focal gains of VEGFA and molecular classification of hepatocellular carcinoma. Cancer Res. 2008;68(16):6779–6788.
Boyault S, Rickman DS, De Reyniès A, et al. Transcriptome classification of HCC is related to gene alterations and to new therapeutic targets. Hepatology. 2007;45(1):42–52.
De Yun F, Hui Z, Tan Y, Cheng RX. Effect of phosphorylation of MAPK and Stat3 and expression of c-fos and c-jun proteins on hepatocarcinogenesis and their clinical significance. World J Gastroenterol. 2001;7(1):33–36.
He S, Zhang J, Zhang W, Chen F, Luo R. Foxa1 inhibits hepatocellular carcinoma progression by suppressing PIK3R1 expression in male patients. J Exp Clin Cancer Res. 2017;36(1):175.
Rodon J, Tabernero J. Improving the armamentarium of PI3K inhibitors with isoform-selective agents: a new light in the darkness. Cancer Discov. 2017;7(7):666–669.
Schuster MB, Porse BT. C/EBPalpha: a tumour suppressor in multiple tissues? Biochim Biophys Acta. 2006;1766(1):88–103.
Costa DB, Li S, Kocher O, et al. Immunohistochemical analysis of C/EBPalpha in non-small cell lung cancer reveals frequent down-regulation in stage II and IIIA tumors: a correlative study of e3590. Lung Cancer (Amsterdam, Netherlands). 2007; 56(1):97–103.
Gery S, Tanosaki S, Bose S, Bose N, Vadgama J, Koeffler HP. Down-regulation and growth inhibitory role of C/EBPalpha in breast cancer. Clin Cancer Res. 2005;11(9):3184–3190.
Bennett KL, Hackanson B, Smith LT, et al. Tumor suppressor activity of CCAAT/enhancer binding protein alpha is epigenetically down-regulated in head and neck squamous cell carcinoma. Cancer Res. 2007;67(10):4657–4664.
Tseng HH, Hwang YH, Yeh KT, Chang JG, Chen YL, Yu HS. Reduced expression of C/EBPalpha protein in hepatocellular carcinoma is associated with advanced tumor stage and shortened patient survival. J Cancer Res Clin Oncol. 2009;135(2): 241–247.
Tomizawa M, Watanabe K, Saisho H, Nakagawara A, Tagawa M. Down-regulated expression of the CCAAT/enhancer binding protein alpha and beta genes in human hepatocellular carcinoma: a possible prognostic marker. Anticancer Res. 2003;23(1a):351–354.
Voutila J, Reebye V, Roberts TC, et al. Development and mechanism of small activating RNA targeting CEBPA, a novel therapeutic in clinical trials for liver cancer. Mol Ther. 2017;25(12): 2705–2714.
Reebye V, Huang KW. Gene activation of CEBPA using saRNA: preclinical studies of the first in human saRNA drug candidate for liver cancer. 2018;37(24):3216-28.
Huang L, Watanabe M, Chikamori M, et al. Unique role of SNT-2/FRS2β/FRS3 docking/adaptor protein for negative regulation in EGF receptor tyrosine kinase signaling pathways. Oncogene. 2006;25(49):6457.
Iejima D, Minegishi Y, Takenaka K, et al. FRS2β, a potential prognostic gene for non-small cell lung cancer, encodes a feedback inhibitor of EGF receptor family members by ERK binding. Oncogene. 2010;29(21):3087.
Turhal N, Bas E, Er O, et al. ERCC1 is not expressed in hepatocellular cancer: a Turkish oncology group, gastrointestinal oncology subgroup study. J Buon. 2010;15(4):794–796.
Cheng L, Spitz MR, Hong WK, Wei Q. Reduced expression levels of nucleotide excision repair genes in lung cancer: a case-control analysis. Carcinogenesis. 2000;21(8):1527–1530.
Simon GR, Sharma S, Cantor A, Smith P, Bepler G. ERCC1 expression is a predictor of survival in resected patients with non-small cell lung cancer. Chest. 2005;127(3):978–983.
Cheng L, Sturgis EM, Eicher SA, Spitz MR, Wei Q. Expression of nucleotide excision repair genes and the risk for squamous cell carcinoma of the head and neck. Cancer. 2002;94(2):393–397.
Ueda S, Shirabe K, Morita K, et al. Evaluation of ERCC1 expression for cisplatin sensitivity in human hepatocellular carcinoma. Ann Surg Oncol. 2011;18(4):1204–1211.
Fautrel A, Andrieux L, Musso O, Boudjema K, Guillouzo A, Langouët S: Overexpression of the two nucleotide excision repair genes ERCC1 and XPC in human hepatocellular carcinoma. J Hepatol. 2005;43(2):288–293.
Lopez PM, Patel P, Uva P, Villanueva A, Llovet JM. Current management of liver cancer. Eur J Cancer Suppl. 2007;5(5):444–446.
Xu XR, Huang J, Xu ZG, Qian BZ, et al. Insight into hepatocellular carcinogenesis at transcriptome level by comparing gene expression profiles of hepatocellular carcinoma with those of corresponding noncancerous liver. Proc Natl Acad Sci USA. 2001;98(26):15089–15094.
Boyault S, Rickman DS, de Reynies A, et al. Transcriptome classification of HCC is related to gene alterations and to new therapeutic targets. Hepatology. 2007;45(1):42–52.
Chuma M, Sakamoto M, Yamazaki K, et al. Expression profiling in multistage hepatocarcinogenesis: identification of HSP70 as a molecular marker of early hepatocellular carcinoma. Hepatology. 2003;37(1):198–207.
Makowska Z, Boldanova T, Adametz D, et al. Gene expression analysis of biopsy samples reveals critical limitations of transcriptome-based molecular classifications of hepatocellular carcinoma. J Pathol Clin Res. 2016;2(2):80–92.
Waxman S, Wurmbach E. De-regulation of common housekeeping genes in hepatocellular carcinoma. BMC Genomics. 2007;8:243.
Zucman-Rossi J, Villanueva A, Nault J-C, Llovet JM. Genetic landscape and biomarkers of hepatocellular carcinoma. Gastroenterology. 2015;149(5):1226–1239. e1224.
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