Upregulation of the oestrogen target gene SIX1 is associated with higher growth speed and decreased survival in HCV‑positive women with hepatocellular carcinoma

Critelli,Rosina Maria; Critelli,Rosina Maria; Milosa,Fabiola; Milosa,Fabiola; Romanzi,Adriana; Romanzi,Adriana; Lasagni,Simone; Lasagni,Simone; Marcelli,Gemma; Marcelli,Gemma; Di Marco,Lorenza; Di Marco,Lorenza; Pivetti,Alessandra; Pivetti,Alessandra; Schepis,Filippo; Schepis,Filippo; Romagnoli,Dante; Romagnoli,Dante; Mancarella,Serena; Mancarella,Serena; Dituri,Francesco; Dituri,Francesco; Martinez-Chantar,Maria-Luz; Martinez-Chantar,Maria-Luz; Giannelli,Gianluigi; Giannelli,Gianluigi; Villa,Erica; Villa,Erica

doi:10.3892/ol.2022.13515

Upregulation of the oestrogen target gene SIX1 is associated with higher growth speed and decreased survival in HCV‑positive women with hepatocellular carcinoma

Authors:
- Rosina Maria Critelli
- Fabiola Milosa
- Adriana Romanzi
- Simone Lasagni
- Gemma Marcelli
- Lorenza Di Marco
- Alessandra Pivetti
- Filippo Schepis
- Dante Romagnoli
- Serena Mancarella
- Francesco Dituri
- Maria-Luz Martinez-Chantar
- Gianluigi Giannelli
- Erica Villa
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Affiliations: Department of Medical Specialties, Gastroenterology Unit, University of Modena and Reggio Emilia and University Hospital of Modena, I‑41124 Modena, Italy, National Institute of Gastroenterology ‘Saverio de Bellis’, Research Hospital, I‑70013 Castellana Grotte, Italy, Liver Disease Laboratory, Centre for Cooperative Research in Biosciences (CIC bioGUNE), 48160 Derio, Spain
Published online on: September 21, 2022 https://doi.org/10.3892/ol.2022.13515
Article Number: 395
Copyright: © Critelli et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The male/female ratio of patients with hepatocellular carcinoma (HCC) is often unbalanced towards the male sex, indicating a sex predisposition for HCC development. A possible explanation may be attributed to different hormonal statuses, including the pro‑inflammatory action of androgens in men and the protective effects of oestrogen against excessive inflammation in women. Although several studies have studied gene expression in patients with HCC, very few have attempted to identify features that could be distinctive between male and female patients. The present study aimed to identify distinctive signalling mechanisms between men and women that may be associated with HCC progression. The present study analysed a detailed microarray database that was obtained from the prospective study of 78 patients with HCC to study gene expression according to sex. In addition, the present study aimed to evaluate whether the differentially expressed genes were known oestrogen targets. Moreover, RNAs from the HCC cohort were evaluated for microRNA (miRNA/miR) expression, and a relationship between miRNA and gene expression according to sex was investigated. One gene, sineoculis homeobox homolog 1 (SIX1), which is known to be an oestrogen target gene, was revealed to be highly upregulated in hepatitis virus C (HCV)‑positive female patients with HCC but not in HCV‑positive male patients. In addition, SIX1 upregulation had a significant relationship with tumour growth speed (assessed as tumour doubling time in two CTs performed 6 weeks apart) and survival (P=0.009 and P=0.042, respectively) in female patients only. Furthermore, SIX1 upregulation was related with miR‑421 and miR‑9‑5p only in male patients; however, in female patients, SIX1 upregulation had a direct relationship with miR‑181b, miR‑503‑5p and miR‑125b (miRNAs with potential oncogenic capacity), and an inverse correlation with miR139‑5p, miR‑26b, let7c‑3p and let7c‑5p (putatively oncosuppressive microRNAs). These data suggested a distinctive model for liver carcinogenesis in HCV‑positive women, with downregulation of protective mechanisms against tumour progression and the activation of potential oncogenes, in relation to the oestrogen target gene SIX1. (IRB10/08_CE_UniRer; ClinicalTrials ID: NCT01657695).

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1	Villa E, Baldini GM, Pasquinelli C, Melegari M, Cariani E, Di Chirico G and Manenti F: Risk factors for hepatocellular carcinoma in Italy. Male sex, hepatitis B virus, non-A non-B infection, and alcohol. Cancer. 62:611–615. 1988. View Article : Google Scholar : PubMed/NCBI
2	Zhang X, El-Serag HB and Thrift AP: Sex and race disparities in the incidence of hepatocellular carcinoma in the United States examined through age-period-cohort analysis. Cancer Epidemiol Biomarkers Prev. 29:88–94. 2020. View Article : Google Scholar : PubMed/NCBI
3	McGlynn KA, Petrick JL and El-Serag HB: Epidemiology of hepatocellular carcinoma. Hepatology. 73 (Suppl 1):S4–S13. 2021. View Article : Google Scholar : PubMed/NCBI
4	Ma WL, Lai HC, Yeh S, Cai X and Chang C: Androgen receptor roles in hepatocellular carcinoma, cirrhosis, and hepatitis. Endocr Relat Cancer. 21:R165–R182. 2014. View Article : Google Scholar : PubMed/NCBI
5	Li Y, Xu A, Jia S and Huang J: Recent advances in the molecular mechanism of sex disparity in hepatocellular carcinoma. Oncol Lett. 17:4222–4228. 2019.PubMed/NCBI
6	Villa E: Role of estrogen in liver cancer. Womens Health (Lond). 4:41–50. 2008. View Article : Google Scholar : PubMed/NCBI
7	Shi L, Feng Y, Lin H, Ma R and Cai X: Role of estrogen in hepatocellular carcinoma: Is inflammation the key? J Transl Med. 12:932014. View Article : Google Scholar : PubMed/NCBI
8	Naugler WE, Sakurai T, Kim S, Maeda S, Kim K, Elsharkawy AM and Karin M: Gender disparity in liver cancer due to sex differences in MyD88-dependent IL-6 production. Science. 317:121–124. 2007. View Article : Google Scholar : PubMed/NCBI
9	Villa E, Critelli R, Lei B, Marzocchi G, Cammà C, Giannelli G, Pontisso P, Cabibbo G, Enea M, Colopi S, et al: Neoangiogenesis-related genes are hallmarks of fast-growing hepatocellular carcinomas and worst survival. Results from a prospective study. Gut. 65:861–869. 2016. View Article : Google Scholar : PubMed/NCBI
10	Berglund JA, Voelker R, Barber P, Diegel J, Mahady A and Bodner M: RNA regulation by estrogen. Oregon Univ Eugene; 2011, View Article : Google Scholar
11	Li H, Han D, Hou Y, Chen H and Chen Z: Statistical inference methods for two crossing survival curves: A comparison of methods. PLoS One. 10:e01167742015. View Article : Google Scholar : PubMed/NCBI
12	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
13	Martik ML and McClay DR: Deployment of a retinal determination gene network drives directed cell migration in the sea urchin embryo. Elife. 4:e088272015. View Article : Google Scholar : PubMed/NCBI
14	Liu Y, Han N, Zhou S, Zhou R, Yuan X, Xu H, Zhang C, Yin T and Wu K: The DACH/EYA/SIX gene network and its role in tumor initiation and progression. Int J Cancer. 138:1067–1075. 2016. View Article : Google Scholar : PubMed/NCBI
15	Micalizzi DS, Christensen KL, Jedlicka P, Coletta CD, Barón AE, Harrel JC, Horwitz KB, Billheimer D, Heichman KA, Welm AL, et al: The Six1 homeoprotein induces human mammary carcinoma cells to undergo epithelial-mesenchymal transition and metastasis in mice through increasing TGF-beta signaling. J Clin Invest. 119:2678–2690. 2009. View Article : Google Scholar : PubMed/NCBI
16	Micalizzi DS, Wang CA, Farabaugh SM, Schiemann WP and Ford HL: Homeoprotein Six1 increases TGF-beta type I receptor and converts TGF-beta signaling from suppressive to supportive for tumor growth. Cancer Res. 70:10371–10380. 2010. View Article : Google Scholar : PubMed/NCBI
17	Min WP and Wei XF: Silencing SIX1 inhibits epithelial mesenchymal transition through regulating TGF-β/Smad2/3 signaling pathway in papillary thyroid carcinoma. Auris Nasus Larynx. 48:487–495. 2021. View Article : Google Scholar : PubMed/NCBI
18	Liu W, Gao M, Li L, Chen Y, Fan H, Cai Q, Shi Y, Pan C, Liu J, Cheng L, et al: Homeoprotein SIX1 compromises antitumor immunity through TGF-β-mediated regulation of collagens. Cell Mol Immunol. 18:2660–2672. 2021. View Article : Google Scholar : PubMed/NCBI
19	Critelli R, Milosa F, Faillaci F, Condello R, Turola E, Marzi L, Lei B, Dituri F, Andreani S, Sighinolfi P, et al: Microenvironment inflammatory infiltrate drives growth speed and outcome of hepatocellular carcinoma: A prospective clinical study. Cell Death Dis. 8:e30172017. View Article : Google Scholar : PubMed/NCBI
20	Christensen KL, Patrick AN, McCoy EL and Ford HL: The six family of homeobox genes in development and cancer. Adv Cancer Res. 101:93–126. 2008. View Article : Google Scholar : PubMed/NCBI
21	Behbakht K, Qamar L, Aldridge CS, Coletta RD, Davidson SA, Thorburn A and Ford HL: Six1 overexpression in ovarian carcinoma causes resistance to TRAIL-mediated apoptosis and is associated with poor survival. Cancer Res. 67:3036–3042. 2007. View Article : Google Scholar : PubMed/NCBI
22	Cheng Q, Ning D, Chen J, Li X, Chen XP and Jiang L: SIX1 and DACH1 influence the proliferation and apoptosis of hepatocellular carcinoma through regulating p53. Cancer Biol Ther. 19:381–390. 2018. View Article : Google Scholar : PubMed/NCBI
23	Ford HL, Kabingu EN, Bump EA, Mutter GL and Pardee AB: Abrogation of the G2 cell cycle checkpoint associated with overexpression of HSIX1: A possible mechanism of breast carcinogenesis. Proc Natl Acad Sci USA. 95:12608–12613. 1998. View Article : Google Scholar : PubMed/NCBI
24	Jin J, Jin T, Quan M, Piao Y and Lin Z: Ezrin overexpression predicts the poor prognosis of gastric adenocarcinoma. Diagn Pathol. 7:1352012. View Article : Google Scholar : PubMed/NCBI
25	Kahlert C, Lerbs T, Pecqueux M, Herpel E, Hoffmeister M, Jansen L, Brenner H, Chang-Claude J, Bläker H, Kloor M, et al: Overexpression of SIX1 is an independent prognostic marker in stage I–III colorectal cancer. Int J Cancer. 137:2104–2113. 2015. View Article : Google Scholar : PubMed/NCBI
26	Kong J, Zhou X, Liu S, Jin T, Piao Y, Liu C and Lin Z: Overexpression of sineoculis homeobox homolog 1 predicts poor prognosis of hepatocellular carcinoma. Int J Clin Exp Pathol. 7:3018–3027. 2014.PubMed/NCBI
27	Chen K, Wei H, Pan J, Chen Z, Pan D, Gao T, Huang J, Huang M, Ou M and Zhong W: Six1 is negatively correlated with poor prognosis and reduces 5-fluorouracil sensitivity via attenuating the stemness of hepatocellular carcinoma cells. Eur J Pharmacol. 861:1725992019. View Article : Google Scholar : PubMed/NCBI
28	Cui Q, Kong D, Li Z, Ahiable P, Wang K, Wu K and Wu G: Dachshund 1 is differentially expressed between male and female breast cancer: A matched case-control study of clinical characteristics and prognosis. Clinical Breast Cancer. 18:e875–e882. 2018. View Article : Google Scholar : PubMed/NCBI
29	Suen AA, Jefferson WN, Wood CE, Padilla-Banks E, Bae-Jump VL and Williams CJ: SIX1 oncoprotein as a biomarker in a model of hormonal carcinogenesis and in human endometrial cancer. Mol Cancer Res. 14:849–858. 2016. View Article : Google Scholar : PubMed/NCBI
30	Jefferson WN, Padilla-Banks E, Phelps JY, Gerrish KE and Williams CJ: Permanent oviduct posteriorization after neonatal exposure to the phytoestrogen genistein. Environ Health Perspect. 119:1575–1582. 2011. View Article : Google Scholar : PubMed/NCBI
31	Jefferson WN, Chevalier DM, Phelps JY, Cantor AM, Padilla-Banks E, Newbold RR, Archer TK, Kinyamu HK and Williams CJ: Persistently altered epigenetic marks in the mouse uterus after neonatal estrogen exposure. Mol Endocrinol. 27:1666–1677. 2013. View Article : Google Scholar : PubMed/NCBI
32	Rossini GP, Baldini GM, Villa E and Manenti F: Characterization of estrogen receptor from human liver. Gastroenterology. 96:1102–1109. 1989. View Article : Google Scholar : PubMed/NCBI
33	Villa E, Vukotic R, Cammà C, Petta S, Di Leo A, Gitto S, Turola E, Karampatou A, Losi L, Bernabucci V, et al: Reproductive status is associated with the severity of fibrosis in women with hepatitis C. PLoS One. 7:e446242012. View Article : Google Scholar : PubMed/NCBI
34	Meng F, Glaser SS, Francis H, DeMorrow S, Han Y, Passarini JD, Stokes A, Cleary JP, Liu X, Venter J, et al: Functional analysis of microRNAs in human hepatocellular cancer stem cells. J Cell Mol Med. 16:160–173. 2012. View Article : Google Scholar : PubMed/NCBI
35	Wang B, Hsu SH, Majumder S, Kutay H, Huang W, Jacob ST and Ghoshal K: TGFbeta-mediated upregulation of hepatic miR-181b promotes hepatocarcinogenesis by targeting TIMP3. Oncogene. 29:1787–1797. 2010. View Article : Google Scholar : PubMed/NCBI
36	Zhou Q, Zheng X, Chen L, Xu B, Yang X, Jiang J and Wu C: Smad2/3/4 pathway contributes to TGF-β-induced MiRNA-181b expression to promote gastric cancer metastasis by targeting Timp3. Cell Physiol Biochem. 39:453–466. 2016. View Article : Google Scholar : PubMed/NCBI
37	Li B, Liu L, Li X and Wu L: miR-503 suppresses metastasis of hepatocellular carcinoma cell by targeting PRMT1. Biochem Biophys Res Commun. 464:982–987. 2015. View Article : Google Scholar : PubMed/NCBI
38	Xiao Z, Shen J, Zhang L, Li M, Hu W and Cho C: Therapeutic targeting of noncoding RNAs in hepatocellular carcinoma: Recent progress and future prospects. Oncol Lett. 15:3395–3402. 2018.PubMed/NCBI
39	Xiao Y, Tian Q, He J, Huang M, Yang C and Gong L: MiR-503 inhibits hepatocellular carcinoma cell growth via inhibition of insulin-like growth factor 1 receptor. Onco Targets Ther. 9:3535–3544. 2016.PubMed/NCBI
40	Yang X, Zang J, Pan X, Yin J, Xiang Q, Yu J, Gan R and Lei X: miR-503 inhibits proliferation making human hepatocellular carcinoma cells susceptible to 5-fluorouracil by targeting EIF4E. Oncol Rep. 37:563–570. 2017. View Article : Google Scholar : PubMed/NCBI
41	Qiu G, Lin Y, Zhang H and Wu D: miR-139-5p inhibits epithelial-mesenchymal transition, migration and invasion of hepatocellular carcinoma cells by targeting ZEB1 and ZEB2. Biochem Biophys Res Commun. 463:315–321. 2015. View Article : Google Scholar : PubMed/NCBI
42	Li J, Fang L, Yu W and Wang Y: MicroRNA-125b suppresses the migration and invasion of hepatocellular carcinoma cells by targeting transcriptional coactivator with PDZ-binding motif. Oncol Lett. 9:1971–1975. 2015. View Article : Google Scholar : PubMed/NCBI
43	Yang J, Li Z, Pang Y, Zhou T, Sun J, Cheng XY and Zheng WV: MicroRNA-139-5p negatively regulates NME1 expression in hepatocellular carcinoma cells. Adv Clin Exp Med. 31:655–670. 2022. View Article : Google Scholar : PubMed/NCBI