Modelling and validating three-dimensional human breast and cancerous human breast tissues in vitro

Article


Zuk, A., Burczynska, B., Li, D., Ghali, L., Dilworth, S. and Wen, S. 2020. Modelling and validating three-dimensional human breast and cancerous human breast tissues in vitro. Clinical Oncology and Research. 3 (4), pp. 1-9. https://doi.org/10.31487/j.COR.2020.04.05
TypeArticle
TitleModelling and validating three-dimensional human breast and cancerous human breast tissues in vitro
AuthorsZuk, A., Burczynska, B., Li, D., Ghali, L., Dilworth, S. and Wen, S.
Abstract

In this study three dimensional (3-D) in vitro models of normal breast and breast cancer tissues were developed to mimic closely the in vivo tissue microenvironment and therefore providing reliable models for in vitro studies as well as testing of novel cancer therapies. Normal and cancerous human breast cell lines were used to construct 3-D artificial tissues, where de-epidermalised dermis (DED) was used as a scaffold for both models. Morphological analyses were conducted using haematoxylin and eosin staining. Biomarkers including keratin 5 and 19 as well as α smooth muscle actin and mucin 1 were used to confirm and validate the reliability of the proposed models using immunohistochemical techniques. Findings suggest that the 3-D in vitro models described in this work can serve as functional models of both human normal and cancerous breast tissues. Multiple structures similar to ducts and lobules of human breast in vivo were observed in 3-D in vitro models by the use of H&E, some breast cancer colonies seen in the cancerous 3-D model were similar to the ducto-lobular structures observed in normal 3-D model of the breast but the former cells were more loosely connected, irregular and largely disorganized. The established 3-D in vitro model of normal breast showed the development of ducto-lobular structures composed of an inner cell layer which was stained positive with α mucin 1 antibody, a biomarker that is characteristic for luminal cells; and also an outer basal layer of cells that was stained positive for α smooth muscle actin, a biomarker of myoepithelial cells.. Keratin staining in 3-D in vitro models also resembled the pattern observed in vivo where keratin 5 was detected in both luminal and myoepithelial cells of normal breast model (NTERT cells), whereas keratin 19 was present in breast cancer model (C2321 cells). These 3-D models successfully recapitulate both normal and pathological tissue architecture of breast tissue and has the potential for various applications in the evaluation of breast cancer progression and treatment.

KeywordsBreast cancer, keratins, alpha smooth muscle actin, mucin 1, three dimensional in vitro models
Research GroupBiomarkers for Cancer group
LanguageEnglish
PublisherScience Repository
JournalClinical Oncology and Research
ISSN2613-4942
Publication dates
Online21 Apr 2020
Print30 Apr 2020
Publication process dates
Deposited30 Apr 2020
Submitted02 Apr 2020
Accepted16 Apr 2020
Output statusPublished
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© 2020 Lucy Ghali, Anna Karolina Zuk. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Hosting by Science Repository. All rights
reserved. http://dx.doi.org/10.31487/j.COR.2020.04.05

Digital Object Identifier (DOI)https://doi.org/10.31487/j.COR.2020.04.05
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Brown, V., Atkins, C., Ghali, L., Cerio, R., Harwood, C. and Proby, C. 2005. Safety and efficacy of 5% imiquimod cream for the treatment of skin dysplasia in high-risk renal transplant recipients: randomized, double-blind, placebo-controlled trial. Archives of Dermatology. 141 (8), pp. 985-993.
Overexpression of the Axl tyrosine kinase receptor in cutaneous SCC-derived cell lines and tumours
Green, J., Ikram, M., Vyas, J., Patel, N., Proby, C., Ghali, L., Leigh, I., O'Toole, E. and Storey, A. 2006. Overexpression of the Axl tyrosine kinase receptor in cutaneous SCC-derived cell lines and tumours. British journal of cancer. 94 (10), pp. 1446-1451.
Role of HPV E6 proteins in preventing UVB-induced release of pro-apoptotic factors from the mitochondria.
Leverrier, S., Bergamaschi, D., Ghali, L., Ola, A., Warnes, G., Akgul, B., Blight, K., Garcia-Escudero, R., Penna, A., Eddaoudi, A. and Storey, A. 2007. Role of HPV E6 proteins in preventing UVB-induced release of pro-apoptotic factors from the mitochondria. Apoptosis: an international journal on programmed cell death. 12 (3), pp. 549-560. https://doi.org/10.1007/s10495-006-0004-1
Role for WNT16B in human epidermal keratinocyte proliferation and differentiation
Teh, M., Blaydon, D., Ghali, L., Briggs, V., Edmunds, S., Pantazi, E., Barnes, M., Leigh, I., Kelsell, D. and Philpott, M. 2007. Role for WNT16B in human epidermal keratinocyte proliferation and differentiation. Journal of Cell Science. 120 (2), pp. 330-339.
Longitudinal muscle shows abnormal relaxation responses to nitric oxide and contains altered levels of NOS1 and elastin in uncomplicated diverticular disease.
Golder, M., Burleigh, D., Ghali, L., Feakins, R., Lunniss, P., Williams, N. and Navsaria, H. 2007. Longitudinal muscle shows abnormal relaxation responses to nitric oxide and contains altered levels of NOS1 and elastin in uncomplicated diverticular disease. Colorectal disease. 9 (3), pp. 218-228.
HPV8 early genes modulate differentiation and cell cycle of primary human adult keratinocytes.
Akgul, B., Ghali, L., Davies, D., Pfister, H., Leigh, I. and Storey, A. 2007. HPV8 early genes modulate differentiation and cell cycle of primary human adult keratinocytes. Clinical and experimental dermatology. 16 (7), pp. 590-599.
Granulosa cell survival and proliferation are altered in polycystic ovary syndrome
Das, M., Djahanbakhch, O., Hacihanefioglu, B., Saridogan, E., Ikram, M., Ghali, L., Raveendran, M. and Storey, A. 2008. Granulosa cell survival and proliferation are altered in polycystic ovary syndrome. Journal of Clinical Endocrinology and Metabolism. 93 (3), pp. 881-887. https://doi.org/10.1210/jc.2007-1650
GLI1 repression of ERK activity correlates with colony formation and impaired migration in human epidermal keratinocytes
Neill, G., Harrison, W., Ikram, M., Williams, T., Bianchi, L., Nadenla, S., Green, J., Ghali, L., Frischauf, A., O'Toole, E., Aberger, F. and Philpott, M. 2008. GLI1 repression of ERK activity correlates with colony formation and impaired migration in human epidermal keratinocytes. Carcinogenesis. 29 (4), pp. 738-746. https://doi.org/10.1093/carcin/bgn037
Follicular fluid levels of inhibin A, inhibin B, and activin A levels reflect changes in follicle size but are not independent markers of the oocyte's ability to fertilize
Wen, S., Tozer, A., Butler, S., Bell, C., Docherty, S. and Iles, R. 2006. Follicular fluid levels of inhibin A, inhibin B, and activin A levels reflect changes in follicle size but are not independent markers of the oocyte's ability to fertilize. Fertility and Sterility. 85, pp. 1723-1729. https://doi.org/10.1016/j.fertnstert.2005.11.058
Gonadotropins and prostate cancer: revisited.
Thwaini, A., Naase, M., Chinegwundon, F., Baithun, S., Ghali, L., Shergill, I. and Iles, R. 2006. Gonadotropins and prostate cancer: revisited. Urologia internationalis. 77 (4), pp. 289-296. https://doi.org/10.1159/000096330
Smooth muscle cholinergic denervation hypersensitivity in diverticular disease.
Ghali, L., Belai, A., Burleigh, D. and Golder, M. 2003. Smooth muscle cholinergic denervation hypersensitivity in diverticular disease. The Lancet. 361 (9373), pp. 1945-1951. https://doi.org/10.1016/S0140-6736(03)13583-0
Epidermal and hair follicle progenitor cells express melanoma-associated chondroitin sulfate proteoglycan core protein
Ghali, L., Wong, S., Tidman, N., Quinn, A., Philpott, M. and Leigh, I. 2004. Epidermal and hair follicle progenitor cells express melanoma-associated chondroitin sulfate proteoglycan core protein. Journal of investigative dermatology. 122 (2), pp. 433-442. https://doi.org/10.1046/j.0022-202X.2004.22207.x
FOXM1 is a downstream target of Gli1 in basal cell carcinomas
Teh, M., Wong, S., Neill, G., Ghali, L., Philpott, M. and Quinn, A. 2002. FOXM1 is a downstream target of Gli1 in basal cell carcinomas. Cancer Research. 62 (16), pp. 4773-4780.
Loss of protein kinase calpha expression may enhance the tumorigenic potential of Gli1 in basal cell carcinoma
Ghali, L., Green, J., Ikram, M. and Neill, G. 2003. Loss of protein kinase calpha expression may enhance the tumorigenic potential of Gli1 in basal cell carcinoma. Cancer Research. 63 (15), pp. 4692-4697.
Human granulosa-lutein cell in vitro production of progesterone, inhibin A, inhibin B, and activin A are dependent on follicular size and not the presence of the oocyte
Wen, S., Tozer, A., Li, D., Docherty, S., Al-Shawaf, T. and Iles, R. 2008. Human granulosa-lutein cell in vitro production of progesterone, inhibin A, inhibin B, and activin A are dependent on follicular size and not the presence of the oocyte. Fertility and Sterility. 89 (5), pp. 1406-1413. https://doi.org/10.1016/j.fertnstert.2007.03.086
Characteristics of populations of granulosa cells from individual follicles in women undergoing 'coasting' during controlled ovarian stimulation (COS) for IVF
Tozer, A., Iles, R., Iammarrone, E., Gillott, C., Wen, S., Al-Shawaf, T. and Grudzinskas, J. 2004. Characteristics of populations of granulosa cells from individual follicles in women undergoing 'coasting' during controlled ovarian stimulation (COS) for IVF. Human Reproduction. 19 (11), pp. 2561-2568. https://doi.org/10.1093/humrep/deh487