Uchikado Yasuto, Inoue Hiroshi, Haraguchi Naotsugu, Mimori Koshi, Natsugoe Shoji, Okumura Hiroshi, Aikou Takashi, Mori Masaki
Department of Surgical Oncology, Digestive Surgery, Graduate School of Medicine, Kagoshima University, Kagoshima 890-8520, Japan.
Int J Oncol. 2006 Dec;29(6):1337-47.
We applied oligomicroarray analysis of 17086 genes to identify the genes related to lymph node metastasis in esophageal squamous cell carcinoma (ESCC). The samples of cancer and non-cancerous paired tissue were taken from 16 patients with ESCC who underwent esophagectomy with lymph node dissection. Total ribonucleic acid was extracted from the cancer cells obtained by using laser microdissection and was amplified by T7 based-amplification for the application to the oligomicroarray. The oligomicroarray demonstrated 43 overexpressed genes, such as cell-cycle regulators, cell adhesion related genes, anti-apoptosis related genes, and 138 suppressed genes such as cell differentiation related and apoptosis related genes in ESCC cells with lymph node metastasis. Among them, 5 overexpressed genes (SPP-1, CKS2, CCT5, STMN1, NDUFB9) and one suppressed expression gene (GJB2) were selected in the gene profiles, and then the expressions of those genes were confirmed by real-time semi-quantitative reverse transcriptional polymerase chain reaction (RT-PCR) method for confirmation of the result not only in study cases but also in additional 21 cases. The gene expression by real-time semi-quantitative RT-PCR was in accordance with the microarray data. Although we were able to extract some genes related to nodal metastasis in ESCC, further examination is necessary in other genes as well as the interaction of stromal tissues.
我们应用包含17086个基因的寡核苷酸微阵列分析来鉴定食管鳞状细胞癌(ESCC)中与淋巴结转移相关的基因。癌组织与癌旁配对组织样本取自16例行食管癌切除术并进行淋巴结清扫的ESCC患者。使用激光显微切割技术从癌细胞中提取总核糖核酸,并通过基于T7的扩增进行扩增,以应用于寡核苷酸微阵列。寡核苷酸微阵列显示,在伴有淋巴结转移的ESCC细胞中,有43个基因过表达,如细胞周期调节因子、细胞黏附相关基因、抗凋亡相关基因等,以及138个基因表达受抑制,如细胞分化相关基因和凋亡相关基因。在这些基因中,从基因谱中选取了5个过表达基因(SPP-1、CKS2、CCT5、STMN1、NDUFB9)和1个表达受抑制基因(GJB2),然后通过实时半定量逆转录聚合酶链反应(RT-PCR)方法确认这些基因的表达,不仅在研究病例中,还在另外21例病例中进行了验证。实时半定量RT-PCR检测的基因表达与微阵列数据一致。虽然我们能够提取出一些与ESCC淋巴结转移相关的基因,但对其他基因以及基质组织的相互作用仍需进一步研究。