Nakatani, Rieko1; Shimasue, Asami3; Yamakawa, Noriko3; Watanabe, Mikio3; Iwatani, Yoshinori 3; Hidaka, Yoh2; Takano, Toru1,2
1 Department of Metabolic Medicine, Osaka University Graduate School of Medicine, Osaka, Japan
2 Department of Laboratory Medicine, Osaka University Graduate School of Medicine,Osaka,Japan
3 Division of Health Sciences, Osaka University Graduate School of Medicine,Osaka, Japan
Background/Purpose: Detection and analysis of a small subpopulation of cells such as stem cells or cancer stem cells are keystone in a recent cancer science. We previously established a method to analyze cells collected by fluorescence-activated cell sorting (FACS), named mRNA quantification after FACS (FACS-mQ). In FACS-mQ, cell sorting is performed without RNA degradation, then gene expression profile is analyzed using RNAs extracted from sorted cells. In this study, we analyzed the expression of stemness genes in a rat thyroid cell lines FRTL5 using this method.
Methods: FACS-mQ was performed using thyroglobulin (TG), thyroid transcription factor 1 (TTF1), and Ki-67 staining. After FACS, the expressions of stemness genes, NANOG, SOX2, PROM1, POU5F1, ABCG2, GATA4, and HNF4A were analyzed by RT-PCR.
Results: Three stemness genes, NANOG, ABCG2, and GATA4 were expressed in FRTL5. FRTL5 cells expressed various TG levels among cell populations. The low-TG cell population showed high expression of the stemness genes.
Conclusion: Analysis of FACS-mQ revealed that FRTL5 contains a cell population with high stemness gene expression and less differentiated features. Focusing our research on this TG-underexpressing cell population might be a promising strategy for further understanding of thyroid stem cells or cancer stem cells.