Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on sleep in brain health

LIVE: Thursday 2nd May 2024, 18:00-19:30 (CEST)

Quality sleep is essential for health. But what happens to our brains when sleep patterns are disturbed? Join our experts to explore the interplay between sleep disruption and neurological diseases, and the questions that you need to be asking your patients to help you prevent the harmful effects of sleep deprivation.
ADVANCED SEARCH
Log in
Top
BMC Cancer
Published in: BMC Cancer 1/2013

Open Access 01-12-2013 | Research article

Characterization of in vivo chemoresistant human hepatocellular carcinoma cells with transendothelial differentiation capacities

Authors: Christian Marfels, Miriam Hoehn, Ernst Wagner, Michael Günther

Published in: BMC Cancer | Issue 1/2013

Login to get access

Abstract

Background

Chemotherapeutic treatment of hepatocellular carcinoma often leads to chemoresistance during therapy or upon relapse of tumors. For the development of better treatments a better understanding of biochemical changes in the resistant tumors is needed. In this study, we focus on the characterization of in vivo chemoresistant human hepatocellular carcinoma HUH-REISO established from a metronomically cyclophosphamide (CPA) treated HUH7 xenograft model.

Methods

SCID mice bearing subcutaneous HUH7 tumors were treated i.p. with 75 mg/kg CPA every six days. Tumors were evaluated by immunohistochemistry, a functional blood-flow Hoechst dye assay, and qRT-PCR for ALDH-1, Notch-1, Notch-3, HES-1, Thy-1, Oct-4, Sox-2 and Nanog mRNA levels. Cell lines of these tumors were analyzed by qRT-PCR and in endothelial transdifferentiation studies on matrigel.

Results

HUH-REISO cells, although slightly more sensitive against activated CPA in vitro than parental HUH-7 cells, fully retained their in vivo CPA chemoresistance upon xenografting into SCID mice. Histochemical analysis of HUH-REISO tumors in comparison to parental HUH-7 cells and passaged HUH-PAS cells (in vivo passaged without chemotherapeutic pressure) revealed significant changes in host vascularization of tumors and especially in expression of the tumor-derived human endothelial marker gene PECAM-1/CD31 in HUH-REISO. In transdifferentiation studies with limited oxygen and metabolite diffusion, followed by a matrigel assay, only the chemoresistant HUH-REISO cells exhibited tube formation potential and expression of human endothelial markers ICAM-2 and PECAM-1/CD31. A comparative study on stemness and plasticity markers revealed upregulation of Thy-1, Oct-4, Sox-2 and Nanog in resistant xenografts. Under therapeutic pressure by CPA, tumors of HUH-PAS and HUH-REISO displayed regulations in Notch-1 and Notch-3 expression.

Conclusions

Chemoresistance of HUH-REISO was not manifested under standard in vitro but under in vivo conditions. HUH-REISO cells showed increased pluripotent capacities and the ability of transdifferentiation to endothelial like cells in vitro and in vivo. These cells expressed typical endothelial surface marker and functionality. Although the mechanism behind chemoresistance of HUH-REISO and involvement of plasticity remains to be clarified, we hypothesize that the observed Notch regulations and upregulation of stemness genes in resistant xenografts are involved in the observed cell plasticity.
Appendix
Available only for authorised users
Literature
1.
Farazi PA, DePinho RA: Hepatocellular carcinoma pathogenesis: from genes to environment. Nat Rev Cancer. 2006, 6: 674-687. 10.1038/nrc1934.CrossRefPubMed
2.
Pang R, Poon RT: Angiogenesis and antiangiogenic therapy in hepatocellular carcinoma. Cancer Lett. 2006, 242: 151-167. 10.1016/j.canlet.2006.01.008.CrossRefPubMed
3.
Finn RS, Zhu AX: Targeting angiogenesis in hepatocellular carcinoma: focus on VEGF and bevacizumab. Expert Rev Anticancer Ther. 2009, 9: 503-509. 10.1586/era.09.6.CrossRefPubMed
4.
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: Sorafenib in advanced hepatocellular carcinoma. N Engl J Med. 2008, 359: 378-390. 10.1056/NEJMoa0708857.CrossRefPubMed
5.
Park ST, Jang JW, Kim GD, Park JA, Hur W, Woo HY, Kim JD, Kwon JH, Yoo CR, Bae SH, Choi JY, Yoon SK: Beneficial effect of metronomic chemotherapy on tumor suppression and survival in a rat model of hepatocellular carcinoma with liver cirrhosis. Cancer Chemother Pharmacol. 2009, 65: 1029-1037.CrossRefPubMed
6.
Browder T, Butterfield CE, Kraling BM, Shi B, Marshall B, O’Reilly MS, Folkman J: Antiangiogenic scheduling of chemotherapy improves efficacy against experimental drug-resistant cancer. Cancer Res. 2000, 60: 1878-1886.PubMed
7.
8.
Broxterman HJ, Lankelma J, Hoekman K: Resistance to cytotoxic and anti-angiogenic anticancer agents: similarities and differences. Drug Resist Updat. 2003, 6: 111-127. 10.1016/S1368-7646(03)00026-8.CrossRefPubMed
9.
Emmenegger U, Shaked Y, Man S, Bocci G, Spasojevic I, Francia G, Kouri A, Coke R, Cruz-Munoz W, Ludeman SM, Colvin OM, Kerbel RS: Pharmacodynamic and pharmacokinetic study of chronic low-dose metronomic cyclophosphamide therapy in mice. Mol Cancer Ther. 2007, 6: 2280-2289. 10.1158/1535-7163.MCT-07-0181.CrossRefPubMed
10.
Thoenes L, Hoehn M, Kashirin R, Ogris M, Arnold GJ, Wagner E, Guenther M: In vivo chemoresistance of prostate cancer in metronomic cyclophosphamide therapy. J Proteomics. 2010, 73: 1342-1354. 10.1016/j.jprot.2010.02.019.CrossRefPubMed
11.
Gunther M, Waxman DJ, Wagner E, Ogris M: Effects of hypoxia and limited diffusion in tumor cell microenvironment on bystander effect of P450 prodrug therapy. Cancer Gene Ther. 2006, 13: 771-779. 10.1038/sj.cgt.7700955.CrossRefPubMed
12.
Abe T, Tada M, Shinohara N, Okada F, Itoh T, Hamada J, Harabayashi T, Chen Q, Moriuchi T, Nonomura K: Establishment and characterization of human urothelial cancer xenografts in severe combined immunodeficient mice. Int J Urol. 2006, 13: 47-57. 10.1111/j.1442-2042.2006.01220.x.CrossRefPubMed
13.
Mueller MM, Peter W, Mappes M, Huelsen A, Steinbauer H, Boukamp P, Vaccariello M, Garlick J, Fusenig NE: Tumor progression of skin carcinoma cells in vivo promoted by clonal selection, mutagenesis, and autocrine growth regulation by granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. Am J Pathol. 2001, 159: 1567-1579. 10.1016/S0002-9440(10)62541-2.CrossRefPubMedPubMedCentral
14.
Gu G, Yuan J, Wills M, Kasper S: Prostate cancer cells with stem cell characteristics reconstitute the original human tumor in vivo. Cancer Res. 2007, 67: 4807-4815. 10.1158/0008-5472.CAN-06-4608.CrossRefPubMed
15.
Zhang S, Balch C, Chan MW, Lai HC, Matei D, Schilder JM, Yan PS, Huang TH, Nephew KP: Identification and characterization of ovarian cancer-initiating cells from primary human tumors. Cancer Res. 2008, 68: 4311-4320. 10.1158/0008-5472.CAN-08-0364.CrossRefPubMedPubMedCentral
16.
Hay DC, Sutherland L, Clark J, Burdon T: Oct-4 knockdown induces similar patterns of endoderm and trophoblast differentiation markers in human and mouse embryonic stem cells. Stem Cells. 2004, 22: 225-235. 10.1634/stemcells.22-2-225.CrossRefPubMed
17.
Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP, Guenther MG, Kumar RM, Murray HL, Jenner RG, Gifford DK, Melton DA, Jaenisch R, Young RA: Core transcriptional regulatory circuitry in human embryonic stem cells. Cell. 2005, 122: 947-956. 10.1016/j.cell.2005.08.020.CrossRefPubMedPubMedCentral
18.
Lu JW, Chang JG, Yeh KT, Chen RM, Tsai JJ, Hu RM: Overexpression of Thy1/CD90 in human hepatocellular carcinoma is associated with HBV infection and poor prognosis. Acta Histochem. 2011, 113: 833-838. 10.1016/j.acthis.2011.01.001.CrossRefPubMed
19.
Chadwick N, Zeef L, Portillo V, Fennessy C, Warrander F, Hoyle S, Buckle AM: Identification of novel Notch target genes in T cell leukaemia. Mol Cancer. 2009, 8: 35-10.1186/1476-4598-8-35.CrossRefPubMedPubMedCentral
20.
Kageyama R, Ohtsuka T, Tomita K: The bHLH gene Hes1 regulates differentiation of multiple cell types. Mol Cells. 2000, 10: 1-7. 10.1007/s10059-000-0001-0.CrossRefPubMed
21.
Lawson ND, Scheer N, Pham VN, Kim CH, Chitnis AB, Campos-Ortega JA, Weinstein BM: Notch signaling is required for arterial-venous differentiation during embryonic vascular development. Development. 2001, 128: 3675-3683.PubMed
22.
Lawson ND, Vogel AM, Weinstein BM: Sonic hedgehog and vascular endothelial growth factor act upstream of the Notch pathway during arterial endothelial differentiation. Dev Cell. 2002, 3: 127-136. 10.1016/S1534-5807(02)00198-3.CrossRefPubMed
23.
Grant DS, Tashiro K, Segui-Real B, Yamada Y, Martin GR, Kleinman HK: Two different laminin domains mediate the differentiation of human endothelial cells into capillary-like structures in vitro. Cell. 1989, 58: 933-943. 10.1016/0092-8674(89)90945-8.CrossRefPubMed
24.
Basu GD, Liang WS, Stephan DA, Wegener LT, Conley CR, Pockaj BA, Mukherjee P: A novel role for cyclooxygenase-2 in regulating vascular channel formation by human breast cancer cells. Breast Cancer Res. 2006, 8: R69-10.1186/bcr1626.CrossRefPubMedPubMedCentral
25.
Scavelli C, Nico B, Cirulli T, Ria R, Di Pietro G, Mangieri D, Bacigalupo A, Mangialardi G, Coluccia AM, Caravita T, Molica S, Ribatti D, Dammacco F, Vacca A: Vasculogenic mimicry by bone marrow macrophages in patients with multiple myeloma. Oncogene. 2008, 27: 663-674. 10.1038/sj.onc.1210691.CrossRefPubMed
26.
El Hallani S, Boisselier B, Peglion F, Rousseau A, Colin C, Idbaih A, Marie Y, Mokhtari K, Thomas JL, Eichmann A, Delattre JY, Maniotis AJ, Sanson M: A new alternative mechanism in glioblastoma vascularization: tubular vasculogenic mimicry. Brain. 2010, 133: 973-982. 10.1093/brain/awq044.CrossRefPubMedPubMedCentral
27.
Bussolati B, Grange C, Sapino A, Camussi G: Endothelial cell differentiation of human breast tumour stem/progenitor cells. J Cell Mol Med. 2009, 13: 309-319. 10.1111/j.1582-4934.2008.00338.x.CrossRefPubMed
28.
Bruno S, Bussolati B, Grange C, Collino F, Graziano ME, Ferrando U, Camussi G: CD133+ renal progenitor cells contribute to tumor angiogenesis. Am J Pathol. 2006, 169: 2223-2235. 10.2353/ajpath.2006.060498.CrossRefPubMedPubMedCentral
Metadata
Title
Characterization of in vivo chemoresistant human hepatocellular carcinoma cells with transendothelial differentiation capacities
Authors
Christian Marfels
Miriam Hoehn
Ernst Wagner
Michael Günther
Publication date
01-12-2013
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2013
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-13-176

Other articles of this Issue 1/2013

BMC Cancer 1/2013 Go to the issue

Research article

Low expression of TFPI-2 associated with poor survival outcome in patients with breast cancer

Research article

Targeting both IGF-1R and mTOR synergistically inhibits growth of renal cell carcinoma in vitro

Research article

Inorganic phosphate and the risk of cancer in the Swedish AMORIS study

Research article

Long non-coding RNA MEG3 inhibits NSCLC cells proliferation and induces apoptosis by affecting p53 expression

Research article

Prognostic value of C-reactive protein and neutrophil-to-lymphocyte ratio in patients with hepatocellular carcinoma

Study protocol

A randomised controlled trial of a mindfulness intervention for men with advanced prostate cancer
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits