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Abstract：
BACKGROUND: Thyroid carcinoma (TC) is the most common endocrine system malignancy, and papillary thyroid carcinoma (PTC) accounts for >80% of all TC cases. Nevertheless, PTC pathogenesis is still not fully understood. The aim of the study was to elucidate the role of the FRMD5 protein in the regulation of biological pathways associated with the development of PTC. We imply that the presence of certain genetic aberrations (e.g., BRAF V600E mutation) is associated with the activity of FRMD5.
METHODS: The studies were conducted on TPC1 and BCPAP (BRAF V600E) model PTC-derived cells. Transfection with siRNA was used to deplete the expression of FRMD5. The mRNA expression and protein yield were evaluated using RT-qPCR and Western blot techniques. Proliferation, migration, invasiveness, adhesion, spheroid formation, and survival tests were performed. RNA sequencing and phospho-kinase proteome profiling were used to assess signaling pathways associated with the FRMD5 expressional status.
RESULTS: The obtained data indicate that the expression of FRMD5 is significantly enhanced in BRAF V600E tumor specimens and cells. It was observed that a drop in intracellular yield of FRMD5 results in significant alternations in the migration, invasiveness, adhesion, and spheroid formation potential of PTC-derived cells. Importantly, significant divergences in the effect of FRMD5 depletion in both BRAF-wt and BRAF-mutated PTC cells were observed. It was also found that knockdown of FRMD5 significantly alters the expression of multidrug resistant genes.
CONCLUSIONS: This is the first report highlighting the importance of the FRMD5 protein in the biology of PTCs. The results suggest that the FRMD5 protein can play an important role in controlling the metastatic potential and multidrug resistance of thyroid tumor cells.
METHODS: The studies were conducted on TPC1 and BCPAP (BRAF V600E) model PTC-derived cells. Transfection with siRNA was used to deplete the expression of FRMD5. The mRNA expression and protein yield were evaluated using RT-qPCR and Western blot techniques. Proliferation, migration, invasiveness, adhesion, spheroid formation, and survival tests were performed. RNA sequencing and phospho-kinase proteome profiling were used to assess signaling pathways associated with the FRMD5 expressional status.
RESULTS: The obtained data indicate that the expression of FRMD5 is significantly enhanced in BRAF V600E tumor specimens and cells. It was observed that a drop in intracellular yield of FRMD5 results in significant alternations in the migration, invasiveness, adhesion, and spheroid formation potential of PTC-derived cells. Importantly, significant divergences in the effect of FRMD5 depletion in both BRAF-wt and BRAF-mutated PTC cells were observed. It was also found that knockdown of FRMD5 significantly alters the expression of multidrug resistant genes.
CONCLUSIONS: This is the first report highlighting the importance of the FRMD5 protein in the biology of PTCs. The results suggest that the FRMD5 protein can play an important role in controlling the metastatic potential and multidrug resistance of thyroid tumor cells.
摘要：
背景：甲状腺癌（TC）是最常见的内分泌系统恶性肿瘤，甲状腺乳头状癌（PTC）占所有TC病例的80％以上。然而，PTC的发病机制尚不完全清楚。该研究的目的是阐明FRMD5蛋白在调节与PTC发展相关的生物途径中的作用。我们暗示存在某些遗传畸变(例如，BRAFV600E突变）与FRMD5的活性相关。
方法:对TPC1和BCPAP(BRAFV600E)模型PTC来源的细胞进行研究。用siRNA转染用于耗尽FRMD5的表达。使用RT-qPCR和Western印迹技术评估mRNA表达和蛋白质产量。扩散,迁移,侵入性,附着力,球状体形成,并进行生存测试。RNA测序和磷酸激酶蛋白质组分析用于评估与FRMD5表达状态相关的信号通路。
结果：获得的数据表明，在BRAFV600E肿瘤标本和细胞中，FRMD5的表达显着增强。观察到FRMD5的细胞内产量下降导致迁移的显着变化，侵入性,附着力,和PTC衍生电池的球状体形成潜力。重要的是,在BRAF-wt和BRAF突变的PTC细胞中观察到FRMD5耗尽的效果的显著差异。还发现FRMD5的敲低显著改变多药抗性基因的表达。
结论：这是第一份报告，强调了FRMD5蛋白在PTC生物学中的重要性。结果表明，FRMD5蛋白在控制甲状腺肿瘤细胞的转移潜能和多药耐药性方面可以发挥重要作用。
方法:对TPC1和BCPAP(BRAFV600E)模型PTC来源的细胞进行研究。用siRNA转染用于耗尽FRMD5的表达。使用RT-qPCR和Western印迹技术评估mRNA表达和蛋白质产量。扩散,迁移,侵入性,附着力,球状体形成,并进行生存测试。RNA测序和磷酸激酶蛋白质组分析用于评估与FRMD5表达状态相关的信号通路。
结果：获得的数据表明，在BRAFV600E肿瘤标本和细胞中，FRMD5的表达显着增强。观察到FRMD5的细胞内产量下降导致迁移的显着变化，侵入性,附着力,和PTC衍生电池的球状体形成潜力。重要的是,在BRAF-wt和BRAF突变的PTC细胞中观察到FRMD5耗尽的效果的显著差异。还发现FRMD5的敲低显著改变多药抗性基因的表达。
结论：这是第一份报告，强调了FRMD5蛋白在PTC生物学中的重要性。结果表明，FRMD5蛋白在控制甲状腺肿瘤细胞的转移潜能和多药耐药性方面可以发挥重要作用。
