Skip to main content

Advertisement

SpringerLink
Log in

Trilostane, a 3β-hydroxysteroid dehydrogenase inhibitor, suppresses growth of hepatocellular carcinoma and enhances anti-cancer effects of sorafenib

  • Preclinical Studies
  • Published:
Investigational New Drugs Aims and scope Submit manuscript

Summary

Background Human 3β-hydroxysteroid dehydrogenase type 1 (HSD3B1) is an enzyme associated with steroidogenesis, however its’ role in hepatocellular carcinoma (HCC) biology is unknown. Trilostane is an inhibitor of HSD3B1 and has been tested as a treatment for patients with breast cancer but has not been studied in patients with HCC. Methods and Results The expression of HSD3B1 in HCC tumors in 57 patients were examined. A total of 44 out of 57 tumors (77.2%) showed increased HSD3B1 expression. The increased HSD3B1 in tumors was significantly associated with advanced HCC. In vitro, the knockdown of HSD3B1 expression in Mahlavu HCC cells by a short hairpin RNA (shRNA) led to significant decreases in colony formation and cell migration. The suppression of clonogenicity in the HSD3B1-knockdown HCC cells was reversed by testosterone and 17β-estradiol. Trilostane-mediated inhibition of HSD3B1 in different HCC cells also caused significant inhibition of clonogenicity and cell migration. In subcutaneous HCC Mahlavu xenografts, trilostane (30 or 60 mg/kg, intraperitoneal injection) significantly inhibited tumor growth in a dose-dependent manner. Furthermore, the combination of trilostane and sorafenib significantly enhanced the inhibition of clonogenicity and xenograft growth, surpassing the effects of each drug used alone, with no documented additional toxicity to animals. HSD3B1 blockade was found to suppress the phosphorylation of extracellular signal-regulated kinase (ERK). The decreased ERK phosphorylation was reversed by testosterone or 17b-estradiol. Conclusions Trilostane significantly inhibited the growth of HCC by inhibiting HSD3B1 function and augmenting the efficacy of sorafenib.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Availability of data and material

The clinicopathological data of patients were retrieved from the database of MacKay Memorial Hospital under the approval from the Institutional Review Board of MacKay Memorial Hospital (13MMHIS269). HCC tissues were collected from patients who underwent liver resection in the Department of Surgery, MacKay Memorial Hospital under the informed consent.

References

  1. Global Burden of Disease Cancer C, Fitzmaurice C, Akinyemiju TF, Al Lami FH, Alam T, Alizadeh-Navaei R, Allen C, Alsharif U, Alvis-Guzman N, Amini E, Anderson BO, Aremu O, Artaman A, Asgedom SW, Assadi R, Atey TM, Avila-Burgos L, Awasthi A, Ba Saleem HO, Barac A, Bennett JR, Bensenor IM, Bhakta N, Brenner H, Cahuana-Hurtado L et al (2018) Global, Regional, and National Cancer Incidence, Mortality, Years of Life Lost, Years Lived With Disability, and Disability-Adjusted Life-Years for 29 Cancer Groups, 1990 to 2016: A Systematic Analysis for the Global Burden of Disease Study. JAMA Oncol 4:1553-1568. https://doi.org/10.1001/jamaoncol.2018.2706

  2. Jemal A, Ward EM, Johnson CJ, Cronin KA, Ma J, Ryerson B, Mariotto A, Lake AJ, Wilson R, Sherman RL, Anderson RN, Henley SJ, Kohler BA, Penberthy L, Feuer EJ, Weir HK (2017) Annual Report to the Nation on the Status of Cancer, 1975-2014, Featuring Survival. J Natl Cancer Inst 109. https://doi.org/10.1093/jnci/djx030

  3. Villanueva A (2019) Hepatocellular Carcinoma. N Engl J Med 380:1450-1462. https://doi.org/10.1056/NEJMra1713263

  4. Roayaie S, Jibara G, Tabrizian P, Park JW, Yang J, Yan L, Schwartz M, Han G, Izzo F, Chen M, Blanc JF, Johnson P, Kudo M, Roberts LR, Sherman M (2015) The role of hepatic resection in the treatment of hepatocellular cancer. Hepatology 62:440-451. https://doi.org/10.1002/hep.27745

  5. Ishizawa T, Hasegawa K, Aoki T, Takahashi M, Inoue Y, Sano K, Imamura H, Sugawara Y, Kokudo N, Makuuchi M (2008) Neither multiple tumors nor portal hypertension are surgical contraindications for hepatocellular carcinoma. Gastroenterology 134:1908-1916. https://doi.org/10.1053/j.gastro.2008.02.091

  6. Adnane L, Trail PA, Taylor I, Wilhelm SM (2006) Sorafenib (BAY 43-9006, Nexavar), a dual-action inhibitor that targets RAF/MEK/ERK pathway in tumor cells and tyrosine kinases VEGFR/PDGFR in tumor vasculature. Methods Enzymol 407:597-612. https://doi.org/10.1016/s0076-6879(05)07047-3

  7. Liu L, Cao Y, Chen C, Zhang X, McNabola A, Wilkie D, Wilhelm S, Lynch M, Carter C (2006) Sorafenib blocks the RAF/MEK/ERK pathway, inhibits tumor angiogenesis, and induces tumor cell apoptosis in hepatocellular carcinoma model PLC/PRF/5. Cancer Res 66:11851-11858. https://doi.org/10.1158/0008-5472.can-06-1377

  8. Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A, Schwartz M, Porta C, Zeuzem S, Bolondi L, Greten TF, Galle PR, Seitz JF, Borbath I, Haussinger D, Giannaris T, Shan M, Moscovici M, Voliotis D, Bruix J, Group SIS (2008) Sorafenib in advanced hepatocellular carcinoma. N Engl J Med 359:378-390. https://doi.org/10.1056/NEJMoa0708857

  9. Kudo M, Ueshima K, Yokosuka O, Ogasawara S, Obi S, Izumi N, Aikata H, Nagano H, Hatano E, Sasaki Y, Hino K, Kumada T, Yamamoto K, Imai Y, Iwadou S, Ogawa C, Okusaka T, Kanai F, Akazawa K, Yoshimura KI, Johnson P, Arai Y (2018) Sorafenib plus low-dose cisplatin and fluorouracil hepatic arterial infusion chemotherapy versus sorafenib alone in patients with advanced hepatocellular carcinoma (SILIUS): a randomised, open label, phase 3 trial. Lancet Gastroenterol Hepatol 3:424-432. https://doi.org/10.1016/s2468-1253(18)30078-5

  10. Jung DH, Tak E, Hwang S, Song GW, Ahn CS, Kim KH, Moon DB, Ha TY, Park GC, Ryoo BY, Lee KJ, Kim N, Kwon JH, Jwa EK, Lee SG (2018) Antitumor effect of sorafenib and mammalian target of rapamycin inhibitor in liver transplantation recipients with hepatocellular carcinoma recurrence. Liver Transpl 24:932-945. https://doi.org/10.1002/lt.25191

  11. Thomas JL, Mack VL, Glow JA, Moshkelani D, Terrell JR, Bucholtz KM (2008) Structure/function of the inhibition of human 3beta-hydroxysteroid dehydrogenase type 1 and type 2 by trilostane. J Steroid Biochem Mol Biol 111:66-73. https://doi.org/10.1016/j.jsbmb.2008.04.007

  12. Pham JH, Will CM, Mack VL, Halbert M, Conner EA, Bucholtz KM, Thomas JL (2017) Structure-function relationships for the selective inhibition of human 3beta-hydroxysteroid dehydrogenase type 1 by a novel androgen analog. J Steroid Biochem Mol Biol 174:257-264. https://doi.org/10.1016/j.jsbmb.2017.10.004

  13. Puddefoot JR, Barker S, Vinson GP (2006) Trilostane in advanced breast cancer. Expert Opin Pharmacother 7:2413-2419. https://doi.org/10.1517/14656566.7.17.2413

  14. Chang YC, Chen CK, Chen MJ, Lin JC, Lin CH, Huang WC, Cheng SP, Chen SN, Liu CL (2017) Expression of 3beta-Hydroxysteroid Dehydrogenase Type 1 in Breast Cancer is Associated with Poor Prognosis Independent of Estrogen Receptor Status. Ann Surg Oncol 24:4033-4041. https://doi.org/10.1245/s10434-017-6000-6

  15. Nagasue N, Yu L, Yukaya H, Kohno H, Nakamura T (1995) Androgen and oestrogen receptors in hepatocellular carcinoma and surrounding liver parenchyma: impact on intrahepatic recurrence after hepatic resection. Br J Surg 82:542-547. https://doi.org/10.1002/bjs.1800820435

  16. Pan ZQ, Fang ZQ, Lu WL (2011) Characteristics of gene expression of adrenal cortical steroid synthetase and its regulatory factor in mice with H22 liver cancer of different patterns. Zhongguo Zhong Xi Yi Jie He Za Zhi 31:85–89

    PubMed  Google Scholar 

  17. Bulayeva NN, Gametchu B, Watson CS (2004) Quantitative measurement of estrogen-induced ERK 1 and 2 activation via multiple membrane-initiated signaling pathways. Steroids 69:181-192. https://doi.org/10.1016/j.steroids.2003.12.003

  18. Hearn JWD, Sweeney CJ, Almassi N, Reichard CA, Reddy CA, Li H, Hobbs B, Jarrard DF, Chen YH, Dreicer R, Garcia JA, Carducci MA, DiPaola RS, Sharifi N (2020) HSD3B1 Genotype and Clinical Outcomes in Metastatic Castration-Sensitive Prostate Cancer. JAMA Oncol:e196496. https://doi.org/10.1001/jamaoncol.2019.6496

  19. Hanamura T, Ito T, Kanai T, Maeno K, Shimojo Y, Uehara T, Suzuki T, Hayashi S, Ito K (2016) Human 3beta-hydroxysteroid dehydrogenase type 1 in human breast cancer: clinical significance and prognostic associations. Cancer Med 5:1405-1415. https://doi.org/10.1002/cam4.708

  20. Chou YY, Jeng YM, Mao TL (2013) HSD3B1 is a specific trophoblast-associated marker not expressed in a wide spectrum of tumors. Int J Gynecol Cancer 23:343-347. https://doi.org/10.1097/IGC.0b013e31827eaa78

  21. Potts GO, Creange JE, Hardomg HR, Schane HP (1978) Trilostane, an orally active inhibitor of steroid biosynthesis. Steroids 32:257-267. https://doi.org/10.1016/0039-128x(78)90010-7

  22. Jungmann E, Althoff PH, Balzer-Kuna S, Magnet W, Rottmann-Kuhnke U, Sprey R, Schwedes U, Usadel KH, Schöffling K (1983) The inhibiting effect of trilostane on testosterone synthesis. Hormonal and morphologic alterations induced by subchronic trilostane treatment in rats and healthy volunteers. Arzneimittelforschung 33:754–756

    CAS  PubMed  Google Scholar 

  23. Takizawa I, Nishiyama T, Hara N, Hoshii T, Ishizaki F, Miyashiro Y, Takahashi K (2010) Trilostane, an inhibitor of 3β-hydroxysteroid dehydrogenase, has an agonistic activity on androgen receptor in human prostate cancer cells. Cancer Lett 297:226-230. https://doi.org/10.1016/j.canlet.2010.05.015

  24. Mellett Keith AM, Bruyette D, Stanley S (2013) Trilostane therapy for treatment of spontaneous hyperadrenocorticism in cats: 15 cases (2004-2012). J Vet Intern Med 27:1471-1477. https://doi.org/10.1111/jvim.12178

  25. King JB, Morton JM (2017) Incidence and risk factors for hypoadrenocorticism in dogs treated with trilostane. Vet J 230:24-29. https://doi.org/10.1016/j.tvjl.2017.10.018

  26. Llovet JM, Montal R, Sia D, Finn RS (2018) Molecular therapies and precision medicine for hepatocellular carcinoma. Nat Rev Clin Oncol 15:599-616. https://doi.org/10.1038/s41571-018-0073-4

  27. Ciou SC, Chou YT, Liu YL, Nieh YC, Lu JW, Huang SF, Chou YT, Cheng LH, Lo JF, Chen MJ, Yang MC, Yuh CH, Wang HD (2015) Ribose-5-phosphate isomerase A regulates hepatocarcinogenesis via PP2A and ERK signaling. Int J Cancer 137:104-115. https://doi.org/10.1002/ijc.29361

  28. Minagawa T, Yamazaki K, Masugi Y, Tsujikawa H, Ojima H, Hibi T, Abe Y, Yagi H, Kitago M, Shinoda M, Itano O, Kitagawa Y, Sakamoto M (2020) Activation of extracellular signal-regulated kinase is associated with hepatocellular carcinoma with aggressive phenotypes. Hepatol Res 50:353-364. https://doi.org/10.1111/hepr.13445

  29. Cheng AL, Kang YK, Chen Z, Tsao CJ, Qin S, Kim JS, Luo R, Feng J, Ye S, Yang TS, Xu J, Sun Y, Liang H, Liu J, Wang J, Tak WY, Pan H, Burock K, Zou J, Voliotis D, Guan Z (2009) Efficacy and safety of sorafenib in patients in the Asia-Pacific region with advanced hepatocellular carcinoma: a phase III randomised, double-blind, placebo-controlled trial. Lancet Oncol 10:25-34. https://doi.org/10.1016/s1470-2045(08)70285-7

Download references

Acknowledgement

We thank Miss Yi-Min Liu for performing part of the cell culture experiments. We are grateful to Dr Sheng-Hsiang Li for kindly providing testosterone and estradiol. We also thank the National RNAi Core Facility at Academia Sinica in Taiwan for providing shRNA reagents and related services.

Funding

This work was supported by research grants (MMH-104-89, MMH-106-75, MMH-108-E-10 and MMH-109-E-10) from MacKay Memorial Hospital.

Author information

Authors and Affiliations

  1. Department of Surgery, MacKay Memorial Hospital and Mackay Medical College, New Taipei City, Taiwan

    Jiunn-Chang Lin, Chien-Liang Liu, Yuan-Ching Chang, Shih-Ping Cheng, Wen-Chien Huang & Ming-Jen Chen

  2. Mackay Junior College of Medicine, Nursing, and Management, New Taipei City, Taiwan

    Jiunn-Chang Lin & Yuan-Ching Chang

  3. Department of Medical Research, MacKay Memorial Hospital, New Taipei City, Taiwan

    Chien-Liang Liu, Shih-Ping Cheng, Chi-Hsin Lin, Chun-Yi Wu & Ming-Jen Chen

  4. Department of Bioscience Technology, Chung Yuan Christian University, Taoyuan City, Taiwan

    Chi-Hsin Lin

Authors
  1. Jiunn-Chang Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chien-Liang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuan-Ching Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shih-Ping Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wen-Chien Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chi-Hsin Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Chun-Yi Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ming-Jen Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

MJ Chen and JC Lin contributed to the study concept and design. YC Chang initiated the study examining trilostane for the treatment for various cancer types. CL Liu, SP Cheng, WC Huang provided logistical support. CH Lin and CY Wu helped with data collection and analysis. All authors reviewed and approved the final manuscript.

Corresponding author

Correspondence to Ming-Jen Chen.

Ethics declarations

Ethical approval and consent to participate

In this study the use of human samples from the surgically removed specimen was approved by the Institutional Review Board of MacKay Memorial Hospital (13MMHIS269). Informed consent was obtained from all patients in order to obtain tissue samples. All animal experiments were approved by the Institutional Animal Care and Use Committee of MacKay Memorial Hospital (MMH-A-S-103-46).

Consent for publication

Signed consent was obtained from all the participating patients for the study of the surgically removed tumors and subsequent publication of the results.

Competing interests

The authors declare that they have no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 305 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lin, JC., Liu, CL., Chang, YC. et al. Trilostane, a 3β-hydroxysteroid dehydrogenase inhibitor, suppresses growth of hepatocellular carcinoma and enhances anti-cancer effects of sorafenib. Invest New Drugs 39, 1493–1506 (2021). https://doi.org/10.1007/s10637-021-01132-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10637-021-01132-3

Keywords

Search

Navigation