Maltseva D V, Makarova J A, Khristichenko A Yu, Tsypina I M, Tonevitsky E A, Rodin S A
SRC Bioclinicum, Moscow, 115088 Russia.
Hertsen Moscow Oncology Research Center, Branch of Federal State Budgetary Institution National Medical Research Radiological Center of the Ministry of Healthcare of the Russian Federation, Obninsk, 249036 Russia.
Mol Biol (Mosk). 2019 Mar-Apr;53(2):330-338. doi: 10.1134/S0026898419020113.
Received August 28, 2018; revised October 10, 2018; accepted November 6, 2018 The loss of apical-basal cell polarity is a necessary stage of the epithelial-mesenchymal transition (EMT). Polarized epithelial cells interact with the basement membrane (BM) and, in particular, with laminins, the major components of BM. Here, we examined the effect of the transition of colon cancer cells from 2D polarized state to non-polarized 3D state on the expression of EMT associated genes, as well as the role of laminins 332 and 411 (LM-332 and LM-411) in this process. The three studied cell lines, HT-29, HCT-116 and RKO, were found to have different sensitivity to cultivation conditions (2D to 3D changes) and to addition of laminins. One of the possible reasons for this maybe a difference in the initial 2D state of the cells. In particular, it was shown that the cell lines were at different EMT stages. HT-29 exhibited more epithelial expression profile, RKO was more mesenchymal, and HCT-116 was in an intermediate state. The most laminin-sensitive cell line was HCT-116. The magnitude and the specificity of cell response to LM-332 and LM-411 depended on the expression pattern of laminins' receptors. EMT gene expression profile was not substantially changed neither during the transition from 2D to 3D state, nor the presence of laminins' isoforms. However, we detected changes in expression of SNAI1 and ZEB1 genes encoding transcription factors that control the EMT process. Notably, in all three studied cell lines, the expression of SNAI1 was enhanced in response to laminin treatment.
2018年8月28日收到；2018年10月10日修订；2018年11月6日接受 顶-基细胞极性丧失是上皮-间质转化（EMT）的一个必要阶段。极化上皮细胞与基底膜（BM）相互作用，尤其是与BM的主要成分层粘连蛋白相互作用。在此，我们研究了结肠癌细胞从二维极化状态转变为非极化三维状态对EMT相关基因表达的影响，以及层粘连蛋白332和411（LM-332和LM-411）在此过程中的作用。研究的三种细胞系HT-29、HCT-116和RKO对培养条件（二维到三维变化）和层粘连蛋白添加的敏感性不同。造成这种情况的一个可能原因可能是细胞初始二维状态的差异。特别是，结果表明这些细胞系处于不同的EMT阶段。HT-29表现出更多的上皮表达谱，RKO更具间充质特性，而HCT-116处于中间状态。对层粘连蛋白最敏感的细胞系是HCT-116。细胞对LM-332和LM-411反应的程度和特异性取决于层粘连蛋白受体的表达模式。EMT基因表达谱在从二维状态转变为三维状态的过程中以及层粘连蛋白异构体存在的情况下均未发生实质性变化。然而，我们检测到了编码控制EMT过程的转录因子的SNAI1和ZEB1基因表达的变化。值得注意的是，在所有三种研究的细胞系中，SNAI1的表达在层粘连蛋白处理后均增强。
