Oka Ryota, Nakashiro Koh-Ichi, Goda Hiroyuki, Iwamoto Kazuki, Tokuzen Norihiko, Hamakawa Hiroyuki
Department of Oral and Maxillofacial Surgery, Ehime University Graduate School of Medicine, Toon, Ehime 791-0295, Japan.
Oncotarget. 2016 Jan 26;7(4):4882-9. doi: 10.18632/oncotarget.6639.
Cervical lymph node metastasis is an important prognostic factor in oral squamous cell carcinoma (OSCC), but its accurate assessment after sentinel node biopsy or neck dissection is often limited to the histopathological examination of only one or two sections. Previous our study showed the usefulness of the reverse transcription loop-mediated isothermal amplification (RT-LAMP) targeting keratin 19 (KRT19) mRNA for the genetic detection of lymph node metastasis, but the sensitivity was insufficient. Here, we have attempted to identify novel molecular markers for OSCC cells in lymph nodes. We performed microarray analysis to identify genes overexpressed in 7 metastatic lymph nodes from OSCC patients, compared to 1 normal lymph node and 5 salivary glands from non-cancer patients. We then used real-time quantitative RT-PCR (qRT-PCR) and RT-LAMP to compare the expression of these genes in newly resected metastatic and normal lymph nodes. Of 4 genes identified by microarray analysis, annexin A8 (ANXA8) and desmoglein 3 mRNA were detected by qRT-PCR in metastatic lymph nodes but not in normal lymph nodes. Furthermore, ANXA8 mRNA expression was detected in all KRT19-negative metastatic lymph nodes. Both KRT19 and ANXA8 mRNA may be useful markers for detecting lymph node metastases in OSCC patients.
颈部淋巴结转移是口腔鳞状细胞癌(OSCC)重要的预后因素,但其在前哨淋巴结活检或颈部清扫术后的准确评估往往仅限于对一两个切片的组织病理学检查。此前我们的研究显示,针对角蛋白19(KRT19)mRNA的逆转录环介导等温扩增(RT-LAMP)技术在淋巴结转移基因检测方面具有一定作用,但敏感性不足。在此,我们试图鉴定出用于检测淋巴结中OSCC细胞的新型分子标志物。我们进行了微阵列分析,以确定与1个正常淋巴结及5个非癌患者唾液腺相比,在7个OSCC患者转移淋巴结中过表达的基因。随后,我们使用实时定量逆转录聚合酶链反应(qRT-PCR)和RT-LAMP技术,比较这些基因在新切除的转移淋巴结和正常淋巴结中的表达情况。在微阵列分析鉴定出的4个基因中,qRT-PCR检测显示膜联蛋白A8(ANXA8)和桥粒芯糖蛋白3 mRNA在转移淋巴结中可检测到,而在正常淋巴结中未检测到。此外,在所有KRT19阴性的转移淋巴结中均检测到ANXA8 mRNA表达。KRT19和ANXA8 mRNA均可能是检测OSCC患者淋巴结转移的有用标志物。