Tumor Biology

, Volume 35, Issue 8, pp 7603–7610 | Cite as

Retinoic acid receptor-related receptor alpha (RORalpha) is a prognostic marker for hepatocellular carcinoma

  • Rong-Dang Fu
  • Chun-Hui Qiu
  • Hu-An Chen
  • Zhi-Gang Zhang
  • Min-Qiang LuEmail author
Research Article


Retinoic acid receptor-related receptor alpha (RORalpha) has been proven to play a tumor suppressive role in certain types of solid tumors. However, the clinical characteristic of RORalpha has not been reported by far. This study investigated the expression of RORalpha in hepatocellular carcinoma (HCC) and evaluated its relationship with clinical parameters and prognosis in HCC patients. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blot analyses were performed to detect RORalpha expression levels in 20 paired HCC and corresponding adjacent non-cancerous tissues. Immunohistochemistry was performed on 100 archived paraffin-embedded HCC samples. Statistical analyses evaluated the correlations between RORalpha expression and clinicopathological features. qRT-PCR showed that RORalpha mRNA expression was significantly down-regulated in tumors compared to the adjacent non-cancerous tissues, and Western blots found that RORalpha protein expression was also reduced in tumor tissues. Immunohistochemical assays revealed that decreased RORalpha expression was present in 65 % of HCC patients. Correlation analyses showed that RORalpha expression was significantly correlated with serum alpha fetoprotein (AFP, p = 0.005), pathology grade (p < 0.001), tumor recurrence (p = 0.008), and vascular invasion (p < 0.001). Kaplan-Meier analysis revealed that patients with low RORalpha expression levels had a shorter overall and disease-free survival than patients with high expression (p < 0.001 and p = 0.002, respectively). Multivariate regression analysis indicated that RORalpha was an independent predictor for overall survival and disease-free survival. In conclusion, the results of our study showed that down-regulated RORalpha expression was associated with poorer prognosis in HCC patients. RORalpha may be a new potential prognostic marker for HCC patients.


RORalpha NR1F1 Hepatocellular carcinoma Immunohistochemistry Prognosis Biomarker 



This work was supported by the National Natural Science Foundation of China (Grant No. 81170450).

Conflicts of interest



  1. 1.
    Bruix J, Sherman M, Llovet JM, Beaugrand M, Lencioni R, Burroughs AK, et al. Clinical management of hepatocellular carcinoma. Conclusions of the Barcelona-2000 EASL conference. European association for the study of the liver. J Hepatol. 2001;35:421–30.PubMedCrossRefGoogle Scholar
  2. 2.
    He J, Gu D, Wu X, Reynolds K, Duan X, Yao C, et al. Major causes of death among men and women in China. N Engl J Med. 2005;353:1124–34.PubMedCrossRefGoogle Scholar
  3. 3.
    Lee MH, Yang HI, Liu J, Batrla-Utermann R, Jen CL, Iloeje UH, et al. Prediction models of long-term cirrhosis and hepatocellular carcinoma risk in chronic hepatitis B patients: risk scores integrating host and virus profiles. Hepatology. 2013;58:546–54.PubMedCrossRefGoogle Scholar
  4. 4.
    Alisi A, Ghidinelli M, Zerbini A, Missale G, Balsano C. Hepatitis C virus and alcohol: same mitotic targets but different signaling pathways. J Hepatol. 2011;54:956–63.PubMedCrossRefGoogle Scholar
  5. 5.
    Reed CA, Mayhew CN, McClendon AK, Yang X, Witkiewicz A, Knudsen ES. RB has a critical role in mediating the in vivo checkpoint response, mitigating secondary DNA damage and suppressing liver tumorigenesis initiated by aflatoxin B1. Oncogene. 2009;28:4434–43.PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Turati F, Edefonti V, Talamini R, Ferraroni M, Malvezzi M, Bravi F, et al. Family history of liver cancer and hepatocellular carcinoma. Hepatology. 2012;55:1416–25.PubMedCrossRefGoogle Scholar
  7. 7.
    Song P, Feng X, Zhang K, Song T, Ma K, Kokudo N, et al. Screening for and surveillance of high-risk patients with HBV-related chronic liver disease: promoting the early detection of hepatocellular carcinoma in China. Biosci Trends. 2013;7:1–6.PubMedGoogle Scholar
  8. 8.
    Abdel-Rahman O. Systemic therapy for hepatocellular carcinoma (HCC): from bench to bedside. J Egypt Natl Canc Inst. 2013;25:165–71.PubMedCrossRefGoogle Scholar
  9. 9.
    Nault JC, De Reynies A, Villanueva A, Calderaro J, Rebouissou S, Couchy G, et al. A hepatocellular carcinoma 5-gene score associated with survival of patients after liver resection. Gastroenterology. 2013;145:176–87.PubMedCrossRefGoogle Scholar
  10. 10.
    Morris SM, Baek JY, Koszarek A, Kanngurn S, Knoblaugh SE, Grady WM. Transforming growth factor-beta signaling promotes hepatocarcinogenesis induced by p53 loss. Hepatology. 2012;55:121–31.PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Solt LA, Burris TP. Action of RORs and their ligands in (patho)physiology. Trends Endocrinol Metab. 2012;23:619–27.PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Lee JM, Kim IS, Kim H, Lee JS, Kim K, Yim HY, et al. RORalpha attenuates Wnt/beta-catenin signaling by PKCalpha-dependent phosphorylation in colon cancer. Mol Cell. 2010;37:183–95.PubMedCrossRefGoogle Scholar
  13. 13.
    Du J, Xu R. RORalpha, a potential tumor suppressor and therapeutic target of breast cancer. Int J Mol Sci. 2012;13:15755–66.PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Moretti RM, Marelli MM, Motta M, Polizzi D, Monestiroli S, Pratesi G, et al. Activation of the orphan nuclear receptor RORalpha induces growth arrest in androgen-independent DU 145 prostate cancer cells. Prostate. 2001;46:327–35.PubMedCrossRefGoogle Scholar
  15. 15.
    Wang Y, Solt LA, Kojetin DJ, Burris TP. Regulation of p53 stability and apoptosis by a ROR agonist. PLoS One. 2012;7:e34921.PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Chauvet C, Vanhoutteghem A, Duhem C, Saint-Auret G, Bois-Joyeux B, Djian P, et al. Control of gene expression by the retinoic acid-related orphan receptor alpha in HepG2 human hepatoma cells. PLoS One. 2011;6:e22545.PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Moh MC, Lee LH, Yang X, Shen S. HEPN1, a novel gene that is frequently down-regulated in hepatocellular carcinoma, suppresses cell growth and induces apoptosis in HepG2 cells. J Hepatol. 2003;39:580–6.PubMedCrossRefGoogle Scholar
  18. 18.
    Chlenski A, Liu S, Guerrero LJ, Yang Q, Tian Y, Salwen HR, et al. SPARC expression is associated with impaired tumor growth, inhibited angiogenesis and changes in the extracellular matrix. Int J Cancer. 2006;118:310–6.PubMedCrossRefGoogle Scholar
  19. 19.
    Li PD, Zhang WJ, Zhang MY, Yuan LJ, Cha YL, Ying XF, et al. Overexpression of RPS6Kb1 predicts worse prognosis in primary HCC patients. Med Oncol. 2012;29:3070–6.PubMedCrossRefGoogle Scholar
  20. 20.
    Iakova P, Timchenko L, Timchenko NA. Intracellular signaling and hepatocellular carcinoma. Semin Cancer Biol. 2011;21:28–34.PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Kim H, Lee JM, Lee G, Bhin J, Oh SK, Kim K, et al. DNA damage-induced RORalpha is crucial for p53 stabilization and increased apoptosis. Mol Cell. 2011;44:797–810.PubMedCrossRefGoogle Scholar
  22. 22.
    Atorrasagasti C, Malvicini M, Aquino JB, Alaniz L, Garcia M, Bolontrade M, et al. Overexpression of SPARC obliterates the in vivo tumorigenicity of human hepatocellular carcinoma cells. Int J Cancer. 2010;126:2726–40.PubMedGoogle Scholar
  23. 23.
    Pez F, Lopez A, Kim M, Wands JR, Caron DFC, Merle P. Wnt signaling and hepatocarcinogenesis: molecular targets for the development of innovative anticancer drugs. J Hepatol. 2013;59:1107–17.PubMedCrossRefGoogle Scholar
  24. 24.
    Hoshida Y, Nijman SM, Kobayashi M, Chan JA, Brunet JP, Chiang DY, et al. Integrative transcriptome analysis reveals common molecular subclasses of human hepatocellular carcinoma. Cancer Res. 2009;69:7385–92.PubMedCentralPubMedCrossRefGoogle Scholar
  25. 25.
    Monga SP, Mars WM, Pediaditakis P, Bell A, Mule K, Bowen WC, et al. Hepatocyte growth factor induces Wnt-independent nuclear translocation of beta-catenin after met-beta-catenin dissociation in hepatocytes. Cancer Res. 2002;62:2064–71.PubMedGoogle Scholar
  26. 26.
    Ma WJ, Wang HY, Teng LS. Correlation analysis of preoperative serum alpha-fetoprotein (AFP) level and prognosis of hepatocellular carcinoma (HCC) after hepatectomy. World J Surg Oncol. 2013;11:212.PubMedCentralPubMedCrossRefGoogle Scholar
  27. 27.
    Guo HF, Li X, Parker MW, Waltenberger J, Becker PM, Vander KC. Mechanistic basis for the potent anti-angiogenic activity of semaphorin 3F. Biochemistry-Us. 2013;52:7551–8.CrossRefGoogle Scholar
  28. 28.
    Luo J, Tao YM, Luo LB, Yang LY. Expression and prognostic value of SEMA3F protein in hepatocellular carcinoma. Chin J Hepatobiliray Surg. 2011;17:219–22.Google Scholar
  29. 29.
    Xiong G, Wang C, Evers BM, Zhou BP, Xu R. RORalpha suppresses breast tumor invasion by inducing SEMA3F expression. Cancer Res. 2012;72:1728–39.PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Rong-Dang Fu
    • 1
  • Chun-Hui Qiu
    • 1
  • Hu-An Chen
    • 1
  • Zhi-Gang Zhang
    • 2
  • Min-Qiang Lu
    • 1
    Email author
  1. 1.Department of Hepatic SurgeryThe Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhouChina
  2. 2.Department of PathologyThe Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhouChina

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