Peng Chih-Yu, Lin Che-Yi, Chen Szu-Han, Liao Yi-Wen, Yu Cheng-Chia, Lee Shiao-Pieng
School of Dentistry, Chung Shan Medical University, Taichung, Taiwan.
Department of Dentistry, Chung Shan Medical University Hospital, Taichung, Taiwan.
J Dent Sci. 2022 Apr;17(2):718-724. doi: 10.1016/j.jds.2021.11.001. Epub 2021 Nov 26.
BACKGROUND/PURPOSE: Oral cancer has been recognized as one of the most common malignancies worldwide and ranks the fifth leading cause of cancer death in Taiwan. A variety of studies have demonstrated that microRNAs are involved in the regulation of the hallmarks of oral carcinogenesis. Nevertheless, the effect of miR-1266-5p on the tumorigenesis of oral cancer has not been investigated, and not to mention, its functional role in oral cancer.
The upregulation of miR-1266-5p in SASVO3 and SASM5 cells was identified by RNA-Seq and examined by qRT-PCR analysis. The phenotypic assays including proliferation activity, migration capacity, invasion, wound healing, and colony-forming abilities were conducted in oral cancer cells after knockdown of miR-1266-5p. Luciferase reporter and western blotting were used to validate DAB2IP was a direct target of miR-1266-5p in oral cancer.
We identified that miR-1266-5p was significantly overexpressed in highly tumorigenic SASVO3 cells and metastatic SASM5 cells. qRT-PCR revealed that miR-1266 significantly increased upregulated in oral cancer and lymph node metastatic tissues compared to normal counterparts We found that downregulation of miR-1266-5p inhibited the proliferation and clonogenicity capacities of SASVO3 cells. Knockdown of miR-1266-5p also inhibited migration/invasion and self-renewal abilities in SASM5 cells. Moreover, we validated miR-1266-5p directly bound to the 3'UTR of DAB2IP in oral cancer cells. We found that DAB2IP knockdown reversed the inhibitory effects of self-renewal and migration mediated by silencing of miR-1266-5p.
miR-1266 functions as a biomarker in oral cancer patients, and downregulation of miR-1266 may ameliorate the oncogenic and metastasis potential of oral cancer by targeting DAB2IP.
背景/目的：口腔癌已被公认为全球最常见的恶性肿瘤之一，在台湾地区是癌症死亡的第五大主要原因。多项研究表明，微小RNA参与口腔癌发生特征的调控。然而，miR-1266-5p对口腔癌肿瘤发生的影响尚未得到研究，更不用说其在口腔癌中的功能作用了。
通过RNA测序在SASVO3和SASM5细胞中鉴定出miR-1266-5p的上调，并通过qRT-PCR分析进行检测。在敲低miR-1266-5p后的口腔癌细胞中进行包括增殖活性、迁移能力、侵袭、伤口愈合和集落形成能力在内的表型分析。使用荧光素酶报告基因检测和蛋白质印迹法验证DAB2IP是口腔癌中miR-1266-5p的直接靶标。
我们发现miR-1266-5p在高致瘤性的SASVO3细胞和转移性的SASM5细胞中显著过表达。qRT-PCR显示，与正常组织相比，miR-1266在口腔癌和淋巴结转移组织中显著上调。我们发现下调miR-1266-5p可抑制SASVO3细胞的增殖和克隆形成能力。敲低miR-1266-5p也抑制了SASM5细胞的迁移/侵袭和自我更新能力。此外，我们验证了miR-1266-5p在口腔癌细胞中直接与DAB2IP的3'UTR结合。我们发现敲低DAB2IP可逆转由miR-1266-5p沉默介导的自我更新和迁移的抑制作用。
miR-1266在口腔癌患者中作为一种生物标志物发挥作用，下调miR-1266可能通过靶向DAB2IP改善口腔癌的致癌和转移潜能。
