Epithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.
Epithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.
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DOI：
10.1371/journal.pone.0021548
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Rocco G
中科院分区：
文献类型：
--
作者：
Pirozzi G;Tirino V;Camerlingo R;Franco R;La Rocca A;Liguori E;Martucci N;Paino F;Normanno N;Rocco G
癌症干细胞（CSCs）假说认为，只有肿瘤内的一小部分细胞能够启动和维持肿瘤。上皮-间充质转化（EMT）是一种胚胎发育程序，通常在癌症侵袭和转移期间被激活。本研究的目的是利用LC 31肺癌原代细胞系，探讨EMT与CSCs的关系。用2ng/ml TGFβ-1分别处理A549和LC 31细胞30天和80天。为了评估EMT，通过光学显微镜、免疫荧光和细胞计数法评估以下标志物的形态学变化：细胞角蛋白、e-钙粘蛋白、CD 326（上皮标志物）和CD 90以及波形蛋白（间充质标志物）。此外，还进行了Slug、Twist和β-catenin基因的RT-PCR。在TGFβ-1处理和未处理的LC 31细胞系上，我们进行了干细胞性测试，例如肺球生长和干细胞标志物表达，例如Oct 4、Nanog、Sox 2、c-kit和CD 133。使用NOD/SCID小鼠进行CD 133的Western印迹和致瘤性测定。TGFβ-1处理的LC 31细胞系失去其上皮形态，呈现成纤维细胞样外观。对于A549细胞系（作为对照）获得了相同的结果。免疫荧光和流式细胞仪检测结果显示，TGFβ 1处理后LC 31和A549细胞Vimentin和CD 90表达上调，而cytocheratin、e-cadherin和CD 326表达下调。TGFβ-1处理的LC 31细胞株中Slug、Twist和β-catenin mRNA转录水平上调，证实了EMT的发生。该细胞系还显示出Oct 4、Nanog、Sox 2和CD 133的过表达，所有这些基因都是干性基因。此外，TGFβ 1处理的LC 31细胞系可增加NOD/SCID小鼠的肺形成能力和肿瘤形成能力。在原代肺癌细胞系中，通过TGFβ-1暴露诱导EMT导致获得间充质谱和干细胞标志物的表达。
Cancer Stem Cells (CSCs) hypothesis asserts that only a small subset of cells within a tumour is capable of both tumour initiation and sustainment. The Epithelial-Mesenchymal Transition (EMT) is an embryonic developmental program that is often activated during cancer invasion and metastasis. The aim of this study is to shed light on the relationship between EMT and CSCs by using LC31 lung cancer primary cell line. A549 and LC31 cell lines were treated with 2 ng/ml TGFβ-1 for 30 days, and 80 days, respectively. To evaluate EMT, morphological changes were assessed by light microscopy, immunofluorescence and cytometry for following markers: cytokeratins, e-cadherin, CD326 (epithelial markers) and CD90, and vimentin (mesenchymal markers). Moreover, RT-PCR for Slug, Twist and β-catenin genes were performed. On TGFβ-1 treated and untreated LC31 cell lines, we performed stemness tests such as pneumospheres growth and stem markers expression such as Oct4, Nanog, Sox2, c-kit and CD133. Western Blot for CD133 and tumorigenicity assays using NOD/SCID mice were performed. TGFβ-1 treated LC31 cell line lost its epithelial morphology assuming a fibroblast-like appearance. The same results were obtained for the A549 cell line (as control). Immunofluorescence and cytometry showed up-regulation of vimentin and CD90 and down-regulation of cytocheratin, e-cadherin and CD326 in TGFβ-1 treated LC31 and A549 cell lines. Slug, Twist and β-catenin m-RNA transcripts were up-regulated in TGFβ-1 treated LC31 cell line confirming EMT. This cell line showed also over-expression of Oct4, Nanog, Sox2 and CD133, all genes of stemness. In addition, in TGFβ-1 treated LC31 cell line, an increased pneumosphere-forming capacity and tumours-forming ability in NOD/SCID mice were detectable. The induction of EMT by TGFβ-1 exposure, in primary lung cancer cell line results in the acquisition of mesenchymal profile and in the expression of stem cell markers.
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影响因子：
--
作者：
Hayashida T;Jinno H;Kitagawa Y;Kitajima M
通讯作者：
Kitajima M
影响因子：
12.4
作者：
Eramo, A.;Lotti, F.;De Maria, R.
通讯作者：
De Maria, R.
影响因子：
3.7
作者：
Levina V;Marrangoni AM;DeMarco R;Gorelik E;Lokshin AE
通讯作者：
Lokshin AE
影响因子：
11.2
作者：
Casas E;Kim J;Bendesky A;Ohno-Machado L;Wolfe CJ;Yang J
通讯作者：
Yang J
影响因子：
11.2
作者：
Locke, M;Heywood, M;Mackenzie, IC
通讯作者：
Mackenzie, IC