Carrera Manoela, Bitu Carolina C, de Oliveira Carine Ervolino, Cervigne Nilva K, Graner Edgard, Manninen Aki, Salo Tuula, Coletta Ricardo D
Department of Oral Diagnosis, School of Dentistry, University of Campinas Piracicaba, São Paulo, Brazil ; Department of Life Sciences, Bahia State University Bahia, Brazil.
Department of Diagnostics and Oral Medicine, Institute of Dentistry, and Medical Research Center, Oulu University Hospital Oulu, Finland.
Int J Clin Exp Pathol. 2015 Apr 1;8(4):3613-23. eCollection 2015.
Although HOX genes are best known for acting in the regulation of important events during embryogenesis, including proliferation, differentiation and migration, alterations in their expression patterns have been frequently described in cancers. In previous studies we analyzed the expression profile of the members of the HOX family of homeobox genes in oral samples of normal mucosa and squamous cell carcinoma (OSCC) and identified differently expressed genes such as HOXA10. The present study aimed to validate the increased expression of HOXA10 in OSCCs, and to investigate the effects arising from its knockdown in OSCC cells. The levels of HOXA10 mRNA were determined in human OSCC samples and cell lines by quantitative PCR, and HOXA10-mediated effects on proliferation, apoptosis, adhesion, epithelial-mesenchymal transition (EMT), migration and invasion were studied in HSC-3 tongue carcinoma cells by using retrovirus-mediated RNA interference. Higher expression of HOXA10 mRNA was observed in OSCC cell lines and in tumor tissues compared to normal controls. HOXA10 knockdown significantly reduced the proliferation of the tumor cells which was accompanied by increased levels of p21. HOXA10 silencing also significantly induced the expression of EMT markers and enhanced the adhesion, migration and invasion of HSC-3 cells. No effects on cell death were observed after HOXA10 knockdown. The results of the current study confirm the overexpression of HOXA10 in OSCCs, and further demonstrate that its expression is functionally associated with several important biological processes related to oral tumorigenesis, such as proliferation, migration and invasion.
尽管HOX基因在胚胎发育过程中调控重要事件（包括增殖、分化和迁移）方面最为人所知，但在癌症中其表达模式的改变也屡有报道。在先前的研究中，我们分析了口腔正常黏膜和鳞状细胞癌（OSCC）样本中同源盒基因HOX家族成员的表达谱，并鉴定出了如HOXA10等表达差异基因。本研究旨在验证HOXA10在OSCC中的表达增加，并研究其在OSCC细胞中敲低后的影响。通过定量PCR测定人OSCC样本和细胞系中HOXA10 mRNA的水平，并利用逆转录病毒介导的RNA干扰在HSC-3舌癌细胞中研究HOXA10对增殖、凋亡、黏附、上皮-间质转化（EMT）、迁移和侵袭的影响。与正常对照相比，在OSCC细胞系和肿瘤组织中观察到HOXA10 mRNA的表达更高。HOXA10敲低显著降低了肿瘤细胞的增殖，同时伴随着p21水平的升高。HOXA10沉默还显著诱导EMT标志物的表达，并增强了HSC-3细胞的黏附、迁移和侵袭。HOXA10敲低后未观察到对细胞死亡的影响。本研究结果证实了HOXA10在OSCC中的过表达，并进一步证明其表达与口腔肿瘤发生相关的几个重要生物学过程（如增殖、迁移和侵袭）在功能上相关。
