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Cancer Cell International
Published in: Cancer Cell International 1/2017

Open Access 01-12-2017 | Primary Research

Alginate-embedded HuH-7 cells increase MMP-9 and reduce OCLN expression in vitro

Authors: Virginia Andrea Angiolini, Carolina Uribe Cruz, Mónica Luján López, Laura Simon, Ursula Matte

Published in: Cancer Cell International | Issue 1/2017

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Abstract

Background

Hepatocellular carcinoma is a common cancer, ranking third in cancer-associated deaths. An important cause of cancer patients’ mortality is metastasis. At the start of metastasis progression, there is an epithelial-mesenchymal transition, characterized by matrix degradation, junction reductions and vessels formation. HuH-7 is a cell line used in research as an in vitro model for hepatocellular carcinoma. It is known that two-dimensional growth reflects tumor characteristics poorly. In contrast, three-dimensional cultures provide a better approach to the study of tumorigenic potential. The purpose of this work was to mimic a three-dimensional environment in order to assess gene expression of some epithelial-mesenchymal transition and metastasis progression markers in HuH-7 cells and compare them with traditional two-dimensional culture model.

Methods

HuH-7 cells were encapsulated in sodium alginate (three-dimensional model) to be compared with cells grown in two-dimensional flasks. After 4 days in culture, gene expression of Matrix metallopeptidase 9, Occludin, p65, Intercellular adhesion molecule 1 and Vascular endothelial growth factor A was analyzed by qPCR and cytoskeleton assessment was performed by rhodamine-phalloidin staining.

Results

Differences were found in gene expression, with a high increment of Matrix metallopeptidase 9 and Occludin reduction. The cytoskeleton morphology also showed differences, with a cytoplasm restricted only near the nuclei in the three-dimensional model.

Conclusions

This work shows the effects of using sodium alginate capsules as a three-dimensional model to the study of HuH-7. Cells in this 3D system show key markers of epithelial-mesenchymal transition, such as Matrix metallopeptidase 9 overexpression and Occludin down-regulation.
Literature
1.
2.
Davis GL, Dempster J, Meler JD, Orr DW, Walberg MW, Brown B, et al. Hepatocellular carcinoma: management of an increasingly common problem. Proc (Bayl Univ Med Cent). 2008;21:266–80.
3.
4.
Nakabayashi H, Taketa K, Miyano K, Yamane T, Sato J. Growth of human hepatoma cell lines with differentiated functions in chemically defined medium. Cancer Res. 1982;42:3858–63.PubMed
5.
Hung JH, Teng YN, Wang LHC, Su IJ, Wang CCC, Huang W, et al. Induction of Bcl-2 expression by hepatitis B virus pre-S2 mutant large surface protein resistance to 5-fluorouracil treatment in Huh-7 cells. PLoS ONE. 2011;6:e28977.CrossRefPubMedPubMedCentral
6.
Krelle AC, Okoli AS, Mendz GL. Huh-7 human liver cancer cells: a model system to understand hepatocellular carcinoma and therapy. J Cancer Ther. 2013;04:606–31.CrossRef
7.
Basu S, Bhattacharyya SN. Insulin-like growth factor-1 prevents miR-122 production in neighbouring cells to curtail its intercellular transfer to ensure proliferation of human hepatoma cells. Nucleic Acids Res. 2014;42:7170–85.CrossRefPubMedPubMedCentral
8.
Mehta G, Hsiao AY, Ingram M, Luker GD, Takayama S. Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy. J Control Release. 2012;164:192–204.CrossRefPubMedPubMedCentral
9.
Greshock J, Nathanson K, Martin A-M, Zhang L, Coukos G, Weber BL, et al. Cancer cell lines as genetic models of their parent histology: analyses based on array comparative genomic hybridization. Cancer Res. 2007;67:3594–600.CrossRefPubMed
10.
Kimlin LC, Casagrande G, Virador VM. In vitro three-dimensional (3D) models in cancer research: an update. Mol Carcinog. 2013;52(3):167–82.CrossRefPubMed
11.
Dunne LW, Huang Z, Meng W, Fan X, Zhang N, Zhang Q, et al. Human decellularized adipose tissue scaffold as a model for breast cancer cell growth and drug treatments. Biomaterials. 2014;35(18):4940–9.CrossRefPubMed
12.
Haycock JW. 3D cell culture: a review of current approaches and techniques. Methods Mol Biol. 2011;695:1–15.CrossRefPubMed
13.
Jacob A, Prekeris R. The regulation of MMP targeting to invadopodia during cancer metastasis. Front cell Dev Biol. 2015;3(February):4.PubMedPubMedCentral
14.
Mehner C, Hockla A, Miller E, Ran S, Radisky DC, Radisky ES. Tumor cell-produced matrix metalloproteinase 9 (MMP-9) drives malignant progression and metastasis of basal-like triple negative breast cancer. Oncotarget. 2014;5:2736–49.CrossRefPubMedPubMedCentral
15.
16.
Ohradanova A, Gradin K, Barathova M, Zatovicova M, Holotnakova T, Kopacek J, et al. Hypoxia upregulates expression of human endosialin gene via hypoxia-inducible factor 2. Br J Cancer. 2008;99:1348–56.CrossRefPubMedPubMedCentral
17.
Lagranha VL, De Carvalho TG, Giugliani R, Matte U. Treatment of MPS I mice with microencapsulated cells overexpressing IDUA: effect of the prednisolone administration. J Microencapsul. 2013;30(4):383–9.CrossRefPubMed
18.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001;25:402–8.CrossRefPubMed
19.
Tran NM, Dufresne M, Duverlie G, Castelain S, Défarge C, Paullier P, et al. An appropriate selection of a 3D alginate culture model for hepatic Huh-7 cell line encapsulation intended for viral studies. Tissue Eng Part A. 2013;19:103–13.CrossRefPubMed
20.
De Vos P, Lazarjani HA, Poncelet D, Faas MM. Polymers in cell encapsulation from an enveloped cell perspective. Adv Drug Deliv Rev. 2014;67-68:15–34.CrossRefPubMed
21.
Shakibaei M, Kraehe P, Popper B, Shayan P, Goel A, Buhrmann C. Curcumin potentiates antitumor activity of 5-fluorouracil in a 3D alginate tumor microenvironment of colorectal cancer. BMC Cancer. 2015;15:250.CrossRefPubMedPubMedCentral
22.
Shteyer E, Ya’acov AB, Zolotaryova L, Sinai A, Lichtenstein Y, Pappo O, et al. Reduced liver cell death using an alginate scaffold bandage: a novel approach for liver reconstruction after extended partial hepatectomy. Acta Biomater. 2014;10:3209–16.CrossRefPubMed
23.
Lan S-F, Starly B. Alginate based 3D hydrogels as an in vitro co-culture model platform for the toxicity screening of new chemical entities. Toxicol Appl Pharmacol. 2011;256(1):62–72.CrossRefPubMed
24.
Mishra DK, Sakamoto JH, Thrall MJ, Baird BN, Blackmon SH, Ferrari M, et al. Human lung cancer cells grown in an ex vivo 3D lung model produce matrix metalloproteinases not produced in 2D culture. PLoS ONE. 2012;7(9):e45308.CrossRefPubMedPubMedCentral
25.
Chimal-Ramírez GK, Espinoza-Sánchez NA, Utrera-Barillas D, Benítez-Bribiesca L, Velázquez JR, Arriaga-Pizano LA, et al. MMP1, MMP9, and COX2 expressions in promonocytes are induced by breast cancer cells and correlate with collagen degradation, transformation-like morphological changes in MCF-10A acini, and tumor aggressiveness. Biomed Res Int. 2013;2013:279505.
26.
Xu X, Farach-Carson MC, Jia X. Three-dimensional in vitro tumor models for cancer research and drug evaluation. Biotechnol Adv. 2014;10:1–13.
27.
Liu S, Li N, Yu X, Xiao X, Cheng K, Hu J, et al. Expression of intercellular adhesion molecule 1 by hepatocellular carcinoma stem cells and circulating tumor cells. Gastroenterology. 2013;144:1031.CrossRefPubMed
28.
Zhu P-P, Yuan S-G, Liao Y, Qin L-L, Liao W-J. High level of intercellular adhesion molecule-1 affects prognosis of patients with hepatocellular carcinoma. World J Gastroenterol. 2015;21:7254–63.PubMedPubMedCentral
29.
Zhang J, Zhang DL, Jiao XL, Dong Q. S100A4 regulates migration and invasion in hepatocellular carcinoma HepG2 cells via NF-KB-dependent MMP-9 signal. Eur Rev Med Pharmacol Sci. 2013;17:2372–82.PubMed
30.
Baldo G, Quoos Mayer F, Burin M, Carrillo-Farga J, Matte U, Giugliani R. Recombinant encapsulated cells overexpressing alpha-L-iduronidase correct enzyme deficiency in human mucopolysaccharidosis type I cells. Cells Tissues Organs. 2012;195:323–9.CrossRefPubMed
Metadata
Title
Alginate-embedded HuH-7 cells increase MMP-9 and reduce OCLN expression in vitro
Authors
Virginia Andrea Angiolini
Carolina Uribe Cruz
Mónica Luján López
Laura Simon
Ursula Matte
Publication date
01-12-2017
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2017
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/s12935-016-0370-x

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