Jung Ji Eun, Lee Joo Young, Park Hae Ryoun, Kang Ji Wan, Kim Yun Hak, Lee Ji Hye
Department of Life Science in Dentistry, School of Dentistry, Pusan National University, Yangsan 50612, Korea.
BK21 FOUR Project, School of Dentistry, Pusan National University, Yangsan 50612, Korea.
J Cancer. 2021 Jul 3;12(17):5296-5309. doi: 10.7150/jca.56138. eCollection 2021.
Non-coding microRNAs (miRNAs) have been proposed to play diverse roles in cancer biology, including epithelial-mesenchymal transition (EMT) crucial for cancer progression. Previous comparative studies revealed distinct expression profiles of miRNAs relevant to tumorigenesis and progression of oral cancer. With putative targets of these miRNAs mostly validated , it remains unclear whether similar miRNA-target relationships exist . In this study, we employed a hybrid approach, utilizing both and human oral cancer cells, to validate projected miRNA-target relationships relevant to EMT. Notably, overexpression of resulted in significant tissue growth in larval wing discs. The RT-PCR analysis successfully validated a subset of its putative targets, including . Subsequent experiments performed in oral cancer cells confirmed conserved targeting of human by . Furthermore, the elevated level of miR-133 and its targeting of was positively correlated with enhanced migrative ability of oral cancer cells treated with LPS, along with the molecular signature of a facilitated EMT process induced by LPS and TGF-β. The analysis on the RNAseq data also revealed a negative correlation between the expression level of and the survival of oral cancer patients. Taken together, our mammal-to--to-mammal approach successfully validates targeting of by miR-133 both and , underlying the promoted EMT phenotypes and potentially influencing the prognosis of oral cancer patients. This hybrid approach will further aid to widen our scope in investigation of intractable human malignancies, including oral cancer.
非编码微小RNA（miRNA）已被提出在癌症生物学中发挥多种作用，包括对癌症进展至关重要的上皮-间质转化（EMT）。先前的比较研究揭示了与口腔癌发生和进展相关的miRNA的不同表达谱。尽管这些miRNA的假定靶标大多已得到验证，但尚不清楚是否存在类似的miRNA-靶标关系。在本研究中，我们采用了一种混合方法，利用果蝇和人类口腔癌细胞来验证与EMT相关的预测miRNA-靶标关系。值得注意的是，miR-133的过表达导致果蝇幼虫翅芽中的组织显著生长。RT-PCR分析成功验证了其一部分假定靶标，包括[具体靶标名称未给出]。随后在口腔癌细胞中进行的实验证实了miR-133对人类[相关基因未给出]的保守靶向作用。此外，miR-133水平的升高及其对[相关基因未给出]的靶向作用与用脂多糖（LPS）处理的口腔癌细胞增强的迁移能力呈正相关，同时伴随着由LPS和转化生长因子-β（TGF-β）诱导的EMT过程的分子特征。对RNAseq数据的分析还揭示了[相关基因未给出]的表达水平与口腔癌患者生存率之间的负相关。综上所述，我们从哺乳动物到果蝇再到哺乳动物的方法成功验证了miR-133在果蝇和人类中对[相关基因未给出]的靶向作用，这是促进EMT表型的基础，并可能影响口腔癌患者的预后。这种混合方法将进一步有助于拓宽我们对包括口腔癌在内的难治性人类恶性肿瘤的研究范围。
