Department of Biochemistry and Molecular Biology, Centre of Postgraduate Medical Education, 01-813 Warsaw, Poland.
Department of Immunohematology, Centre of Postgraduate Medical Education, 01-813 Warsaw, Poland.
Int J Mol Sci. 2019 May 5;20(9):2212. doi: 10.3390/ijms20092212.
The prospero homeobox 1 (PROX1) transcription factor is a product of one of the lymphangiogenesis master genes. It has also been suggested to play a role in carcinogenesis, although its precise role in tumour development and metastasis remains unclear. The aim of this study was to gain more knowledge on the PROX1 function in thyroid tumorigenesis. Follicular thyroid cancer-derived cells-CGTH-W-1-were transfected with PROX1-siRNA (small interfering RNA) and their proliferation, cell cycle, apoptosis and motility were then analysed. The transcriptional signature of depletion was determined using RNA-Sequencing (RNA-Seq) and the expression of relevant genes was further validated using reverse transcriptase quantitative PCR (RT-qPCR), Western blot and immunocytochemistry. PROX1 depletion resulted in a decreased cell motility, with both migratory and invasive potential being significantly reduced. The cell morphology was also affected, while the other studied cancer-related cell characteristics were not significantly altered. RNA-seq analysis revealed significant changes in the expression of transcripts encoding genes involved in both motility and cytoskeleton organization. Our transcriptional analysis of depleted follicular thyroid carcinoma cells followed by functional and phenotypical analyses provide, for the first time, evidence that PROX1 plays an important role in the metastasis of thyroid cancer cells by regulating genes involved in focal adhesion and cytoskeleton organization in tumour cells.
同源盒转录因子 1（PROX1）是淋巴管生成主基因之一的产物。它也被认为在肿瘤发生中发挥作用，尽管其在肿瘤发展和转移中的确切作用仍不清楚。本研究旨在更深入地了解 PROX1 在甲状腺肿瘤发生中的功能。用 PROX1-siRNA（小干扰 RNA）转染滤泡甲状腺癌细胞-CGTH-W-1，然后分析其增殖、细胞周期、凋亡和运动能力。使用 RNA 测序（RNA-Seq）确定 耗竭的转录特征，并使用逆转录定量 PCR（RT-qPCR）、Western blot 和免疫细胞化学进一步验证相关基因的表达。PROX1 耗竭导致细胞运动能力下降，迁移和侵袭潜力均显著降低。细胞形态也受到影响，而其他研究的癌症相关细胞特征没有明显改变。RNA-seq 分析显示，编码参与运动和细胞骨架组织的基因的转录物的表达发生了显著变化。我们对耗尽的滤泡甲状腺癌细胞进行的转录分析，以及随后的功能和表型分析，首次提供了证据，表明 PROX1 通过调节参与肿瘤细胞中粘着斑和细胞骨架组织的基因，在甲状腺癌细胞的转移中发挥重要作用。
