Assessing Estrogen-Induced Proliferative Response in an Endometrial Cancer Cell Line Using a Universally Applicable Methodological Guide

Parkes, Christina, Kamal, Areege ORCID: 0000-0001-5239-9699, Valentijn, Anthony J, Alnafakh, Rafah, Gross, Stephane R, Barraclough, Roger ORCID: 0000-0002-7203-1194, Moss, Diana, Kirwan, John and Hapangama, Dharani K ORCID: 0000-0003-0270-0150 (2018) Assessing Estrogen-Induced Proliferative Response in an Endometrial Cancer Cell Line Using a Universally Applicable Methodological Guide. INTERNATIONAL JOURNAL OF GYNECOLOGICAL CANCER, 28 (1). pp. 122-133.

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Abstract

<h4>Objective</h4>Translational endometrial cancer (EC) research benefits from an in vitro experimental approach using EC cell lines. We demonstrated the steps that are required to examine estrogen-induced proliferative response, a simple yet important research question pertinent to EC, and devised a pragmatic methodological workflow for using EC cell lines in experimental models.<h4>Methods</h4>Comprehensive review of all commercially available EC cell lines was carried out, and Ishikawa cell line was selected to study the estrogen responsiveness with HEC1A, RL95-2, and MFE280 cell lines as comparators where appropriate, examining relevant differential molecular (steroid receptors) and functional (phenotype, anchorage-independent growth, hormone responsiveness, migration, invasion, and chemosensitivity) characteristics in 2-dimensional and 3-dimensional cultures in vitro using immunocytochemistry, immunofluorescence, quantitative polymerase chain reaction, and Western blotting. In vivo tumor, formation, and chemosensitivity were also assessed in a chick chorioallantoic membrane model.<h4>Results</h4>Short tandem repeat analysis authenticated the purchased cell lines, whereas gifted cells deviated significantly from the published profile. We demonstrate the importance of prior assessment of the suitability of each cell line for the chosen in vitro experimental technique. Prior establishment of baseline, nonenriched conditions was required to induce a proliferative response to estrogen. The chorioallantoic membrane model was a suitable in vivo multicellular animal model for EC for producing rapid and reproducible data.<h4>Conclusions</h4>We have developed a methodological guide for EC researchers when using endometrial cell lines to answer important translational research questions (exemplified by estrogen-responsive cell proliferation) to facilitate robust data, while saving time and resources.

Item Type:	Article
Uncontrolled Keywords:	Cell lines, Endometrial cancer, Estrogen, AR - androgen receptor, CAM - chorioallantoic membrane, CSFBS - charcoal-stripped fetal bovine serum, E2-17-estradiol, EC - endometrial cancer, ER - estrogen receptor, ISK - Ishikawa, NBF - neutral-buffered formalin, PBS - phosphate-buffered saline, PR - progesterone receptor, STR - short tandem repeat
Depositing User:	Symplectic Admin
Date Deposited:	17 Jan 2018 11:03
Last Modified:	19 Jan 2023 06:53
DOI:	10.1097/IGC.0000000000001121
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3009814