Expression analysis of stemness genes in a rat thyroid cell line FRTL5
Expression analysis of stemness genes in a rat thyroid cell line FRTL5
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大鼠甲状腺细胞系FRTL5干性基因的表达分析
DOI：
10.1055/s-0034-1389924
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
T.
中科院分区：
文献类型：
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作者：
Shimasue;A.;Yamakawa;N.;Watanabe;M.;Hidaka;Y.;Iwatani;Y.;Takano;T.
在最近的再生和癌症科学中，检测和分析细胞的小亚群如干细胞或癌症干细胞被认为是关键技术。然而，在甲状腺中，尚未建立识别干细胞的标记物。我们先前建立了一种新的方法来分析通过荧光激活细胞分选（FACS）收集的细胞，在FACS之后命名为mRNA定量（FACS-mQ）。通过使用该方法，可以通过分析分选细胞的基因表达谱来确定分选细胞的生物学特性。在这项研究中，我们分析了干细胞基因的表达在大鼠甲状腺细胞系FRTL 5使用FACS-mQ。FRTL 5中有NANOG、ABCG 2和GATA 4 3个干性基因表达。流式细胞仪检测FRTL 5细胞中甲状腺球蛋白（TG）的表达。通过FACS-mQ分析具有高或低TG表达的细胞群体。低TG表达的细胞群体显示出干性基因的表达增加。此外，Ki 67阳性细胞显示TG表达增加，这表明具有高TG的细胞增殖迅速。这些结果表明，FRTL 5包含具有高干细胞性基因表达和较少分化特征的细胞群，类似于干细胞。这些细胞可能调节FRTL 5的增殖。
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,ABCG2andGATA4,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.