Modelling and validating three dimensional human normal cervix and cervical cancer tissues in vitro

Zuk, Anna Karolina, Wen, Xuesong ORCID: https://orcid.org/0000-0001-6518-8962, Dilworth, Stephen M. ORCID: https://orcid.org/0000-0001-9815-0004, Li, Dong ORCID: https://orcid.org/0000-0001-9240-4173 and Ghali, Lucy ORCID: https://orcid.org/0000-0003-3410-6615 (2017) Modelling and validating three dimensional human normal cervix and cervical cancer tissues in vitro. Journal of Biomedical Research, 31 (3) . pp. 240-247. ISSN 1674-8301 [Article] (doi:10.7555/JBR.31.20160150)

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Abstract

Objective: The use of three dimensional in vitro systems in cancer research is a promising path for developing effective anticancer therapies. The aim of this study was to engineer a functional 3-D in vitro model of normal and cancerous cervical tissue.

Methods: Normal epithelial and immortalized cervical epithelial carcinoma cell lines were used to construct 3-D artificial normal cervical and cervical cancerous tissues. De-epidermised dermis (DED) was used as a scaffold for both models. Morphological analyses were conducted by using haematoxylin and eosin staining and characteristics of the models were studied by analysing the expression of different structural cytokeratins and differential protein marker Mad1 using immunohistochemical technique.

Results: Haematoxylin and eosin staining results showed that normal cervical tissue had multi epithelial layers while cancerous cervical tissue showed dysplastic changes. Immunohistochemistry staining results revealed that for normal cervix model cytokeratin 10 was expressed in the upper stratified layer of epithelium while cytokeratin 5 was expressed mainly in the middle and basal layer. Cytokeratin 19 was weakly expressed in a few basal cells. Cervical cancer model showed cytokeratin 19 expression in different epithelial layers and weak or no expression for cytokeratin 5 and cytokeratin 10. Mad1 expression was detected in some suprabasal cells.

Conclusions: The 3-D in vitro models showed stratified epithelial layers and expressed the same types and patterns of differentiation marker proteins as seen in corresponding in vivo tissue in either normal cervical or cervical cancerous tissue. Findings imply that they can serve as functional normal and cervical cancer models.

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