Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an, China.
Department of Dermatology, Tangdu Hospital, Fourth Military Medical University, Xi'an, China.
J Cell Physiol. 2019 May;234(5):5354-5361. doi: 10.1002/jcp.27405. Epub 2018 Nov 27.
Cancer cells are considered to have high morphological heterogeneity in human melanoma tissue. Here, we report that epithelial cancer cells are dominant in different development stages of human melanoma tissues. The cellular and molecular mechanisms that maintain melanoma cells in the epithelial state are further investigated in the A2058 cell line. We find that micropore (8 µm) transwell invasion, but not superficial migration in the scratch assay, can induce remarkable morphological changes between epithelial and mesenchymal melanoma cells within 4 days. The morphological switch is associated with dynamic changes of epithelial-mesenchymal transition (EMT) hallmarks E-cadherin and vimentin. Further immunoflurencent staining and co-immunoprecipitation assay showed the uncoupling of the M3 muscarinic acetylcholine receptor (mAChR) and the p75 neurotrophin receptor (p75NTR) in epithelial melanoma cells. Specific knockdown of M3 mAChR by small interfering RNA (siRNA) significantly abrogates the transition of spindle-shaped mesenchymal cells to epithelial cells. Collectively, we report a cellular model of invasiveness-triggered state transition (ITST) in which melanoma cell invasion can induce morphological changes between epithelial and mesenchymal cells. ITST is one of the biological basis for maintaining metastatic melanoma cells in the epithelial state. Furthermore, M3 mAChR receptor-mediated ITST provides a novel therapeutic strategy to inhibit the development of malignant melanoma.
癌细胞被认为在人类黑色素瘤组织中具有高度的形态异质性。在这里，我们报告上皮癌细胞在人类黑色素瘤组织的不同发育阶段占主导地位。在 A2058 细胞系中进一步研究了维持黑色素瘤细胞上皮状态的细胞和分子机制。我们发现微孔（8μm）transwell 侵袭，而不是划痕实验中的浅层迁移，可以在 4 天内诱导上皮和间充质黑色素瘤细胞之间发生显著的形态变化。形态转换与上皮-间充质转化（EMT）标志物 E-钙粘蛋白和波形蛋白的动态变化相关。进一步的免疫荧光染色和共免疫沉淀实验表明，上皮黑色素瘤细胞中 M3 毒蕈碱乙酰胆碱受体（mAChR）和 p75 神经营养素受体（p75NTR）的解偶联。通过小干扰 RNA（siRNA）特异性敲低 M3 mAChR 可显著阻断纺锤形间充质细胞向上皮细胞的转化。总之，我们报告了一种侵袭性触发状态转变（ITST）的细胞模型，其中黑色素瘤细胞侵袭可以诱导上皮和间充质细胞之间的形态变化。ITST 是维持转移性黑色素瘤细胞上皮状态的生物学基础之一。此外，M3 mAChR 受体介导的 ITST 为抑制恶性黑色素瘤的发展提供了一种新的治疗策略。
