Department of Oral Pathology, School of Dentistry, Federal University of Para, Avenida Augusto Correa, 01. Belem, Para, 66075-110, Belém, PA 66073-000, Brazil.
Laboratory of Genetics and Molecular Biology, State University of Pará, Travessa Perebebuí, 2623, Belém, PA 66087-670, Brazil.
Cells. 2020 Jan 8;9(1):157. doi: 10.3390/cells9010157.
Mucoepidermoid carcinoma (MEC) is the most common tumor in the salivary glands, often presenting with recurrence and metastasis due to its high invasive capacity. Metallothionein (MT), a zinc storage protein that supplies this element for protease activity, is probably related to mucoepidermoid carcinoma behavior. This prompted us to characterize a cell line derived from mucoepidermoid carcinoma and to correlate metallothionein expression with transforming growth factor-α (TGF-α), tumor necrosis factor-α (TNF-α) and matrix metalloproteinases (MMPs). Transcriptomic analysis and cytogenetic assays were performed to detect the expression of genes of interest and cellular chromosomal alterations, respectively. MEC cells with a depleted metallothionein 2A () gene were subjected to Western blot to correlate metallothionein expression with growth factors and MMPs. Additionally, cells with depleted MT were subjected to migration and invasion assays. The transcriptomic study revealed reads mapped to cytokeratins 19 and AE1/AE3, α-smooth muscle actin, vimentin, and fibronectin. Cytogenetic evaluation demonstrated structural and numerical alterations, including the translocation t(11;19)(q21;p13), characteristic of MEC. Metallothionein depletion was correlated with the decreased expression of TGF-α and MMP-9, while TNF-α protein levels were augmented. Migration and invasion activity were diminished after metallothionein silencing. Our findings suggest an important role of MT in MEC invasion, through the regulation of proteins involved in this process.
黏液表皮样癌(MEC)是唾液腺最常见的肿瘤,由于其高侵袭性,常表现为复发和转移。金属硫蛋白(MT)是一种锌储存蛋白,可为蛋白酶活性提供这种元素,可能与黏液表皮样癌的行为有关。这促使我们对源自黏液表皮样癌的细胞系进行了表征,并将金属硫蛋白的表达与转化生长因子-α(TGF-α)、肿瘤坏死因子-α(TNF-α)和基质金属蛋白酶(MMPs)相关联。进行了转录组分析和细胞遗传学检测,分别用于检测感兴趣基因的表达和细胞染色体改变。用耗尽金属硫蛋白 2A () 基因的 MEC 细胞进行 Western blot,以将金属硫蛋白的表达与生长因子和 MMP 相关联。此外,还对耗尽 MT 的细胞进行了迁移和侵袭测定。转录组研究显示了与细胞角蛋白 19 和 AE1/AE3、α-平滑肌肌动蛋白、波形蛋白和纤连蛋白相对应的读数。细胞遗传学评估显示了结构和数量的改变,包括特征性的 MEC 易位 t(11;19)(q21;p13)。金属硫蛋白的耗竭与 TGF-α 和 MMP-9 的表达减少相关,而 TNF-α 蛋白水平增加。金属硫蛋白沉默后,迁移和侵袭活性降低。我们的研究结果表明 MT 在 MEC 侵袭中具有重要作用,通过调节参与该过程的蛋白质。