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Exp Clin Endocrinol Diabetes 2015; 123(01): 48-54
DOI: 10.1055/s-0034-1389924
Article
© Georg Thieme Verlag KG Stuttgart · New York

Expression Analysis of Stemness Genes in a Rat Thyroid Cell Line FRTL5

A. Shimasue
1   Department of Laboratory Medicine, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
2   Division of Health Sciences, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
,
N. Yamakawa
1   Department of Laboratory Medicine, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
2   Division of Health Sciences, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
,
M. Watanabe
2   Division of Health Sciences, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
,
Y. Hidaka
1   Department of Laboratory Medicine, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
,
Y. Iwatani
2   Division of Health Sciences, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
,
T. Takano
1   Department of Laboratory Medicine, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
› Author Affiliations
Further Information

Publication History

received 24 June 2014
first decision 07 August 2014

accepted 22 August 2014

Publication Date:
14 October 2014 (online)

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Abstract

Detection and analysis of a small subpopulation of cells such as stem cells or cancer stem cells are recognized to be a key technique in a recent regeneration and cancer science. However, in the thyroid, no marker that identifies stem cells has been established yet. We previously established a novel method to analyze cells collected by fluorescence-activated cell sorting (FACS), named mRNA quantification after FACS (FACS-mQ). By using this method, the biological characteristics of the sorted cells can be determined by analyzing their gene expression profile. In this study, we analyzed the expression of stemness genes in a rat thyroid cell lines FRTL5 using FACS-mQ. 3 stemness genes, NANOG, ABCG2 and GATA4, were expressed in FRTL5. In FRTL5 cells, varied expression of thyroglobulin (TG) among cells was observed by flow cytometry. Cell populations with high or low TG expression were analyzed by FACS-mQ. The cell population with low TG expression showed increased expression of the stemness genes. Furthermore, Ki67-positive cells showed increased expression of TG, which suggested that cells with high TG proliferated rapidly. These results indicated that FRTL5 contains a cell population with high stemness gene expression and less differentiated features, resembling stem cells. These cells might regulate proliferation in FRTL5.

Key words

FACS-mQ - thyroglobulin - TTF1 - flow cytometry - quantitative RT-PCR
 

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