Analysis of biological characterization and metastasis-related genes in cell lines derived from the Primary lesion and lymph node metastasis of a squamous cell carcinoma arising in the mandibular gingiva. Homo sapiens

Controlling metastatic lesions is an important part of improving cancer prognosis, in addition to controlling the primary lesion. There have been many histological examinations of primary and metastatic lesions, but little basic research using cell strains from primary and metastatic lesions belonging to the same patient. In this study, we successfully established a cell strain derived from lower gingival carcinoma (WK2) and a strain derived from secondary cervical lymph node metastasis (WK3F) through primary cultures of tissue from a patient with oral squamous cell carcinoma. We then investigated the biological characteristics of the cancer cell strains from these primary and metastatic lesions, and analyzed metastasis-related genes. Comparing the biological characteristics in vitro showed WK3F to have higher cell proliferation ability and shorter cell doubling time than WK2. It also showed increased cell migratory ability, and high invasive and self-replication abilities. Heterotransplantation into nude mice resulted in high tumor formation rates in the tongue and high metastasis rates in the cervical lymph nodes. Changes in WK2 and WK3F gene expression were comprehensively analyzed with the microarray method. Genes with increased expression in WK3F compared to WK2 were extracted when Z-score ≥ 2.0 and ratio ≥ 5.0, while genes with reduced expression in WK3F compared to WK2 were extracted when Z-score ≤ -2.0 and ratio ≤ 0.2. As a result, differences were found in 604 genes. From these, MAGEC1 (88.0 times), MMP-7 (18.6 times), SNAI1 (6.6 times), MACC1 (6.2 times), and HTRA1 (0.012 times) were selected as metastasis-related candidate genes. The results suggest that these molecules could be important to clarifying the mechanism of metastasis, and could become therapeutic targets. Overall design: We successfully established a cell strain derived from lower gingival carcinoma (WK2) and a strain derived from secondary cervical lymph node metastasis (WK3F) through primary cultures of tissue from a patient with oral squamous cell carcinoma. Changes in WK2 and WK3F gene expression were comprehensively analyzed with the microarray method.

控制肿瘤转移灶是改善癌症预后的重要一环，在控制原发肿瘤病灶的基础上，转移灶的管控同样至关重要。目前已有诸多针对原发灶与转移灶的组织学研究，但针对源自同一患者的原发灶及转移灶细胞株开展的基础研究却较为匮乏。本研究通过对1例口腔鳞状细胞癌患者的肿瘤组织进行原代培养，成功建立两株细胞株：一株源自患者下颌牙龈癌原发灶（命名为WK2），另一株源自继发性颈部淋巴结转移灶（命名为WK3F）。随后本研究对这两株分别来自原发灶与转移灶的癌细胞株的生物学特性进行了分析，并对转移相关基因展开了研究。体外生物学特性对比结果显示，相较于WK2，WK3F具备更强的细胞增殖能力与更短的细胞倍增时间，同时其细胞迁移能力显著提升，且拥有更高的侵袭与自我复制能力。裸鼠异种移植实验结果表明，WK3F在舌部具有较高的成瘤率，且可在颈部淋巴结形成高转移率的转移灶。本研究采用基因芯片（microarray）技术对WK2与WK3F的基因表达谱进行了全面分析。以Z评分≥2.0且表达倍数≥5.0为筛选标准，提取WK3F中相较于WK2表达上调的基因；以Z评分≤-2.0且表达倍数≤0.2为筛选标准，提取WK3F中相较于WK2表达下调的基因。最终共筛选得到604个差异表达基因，其中包括MAGEC1（上调88.0倍）、MMP-7（上调18.6倍）、SNAI1（上调6.6倍）、MACC1（上调6.2倍）以及HTRA1（下调至原表达量的0.012倍），上述基因被选为转移相关候选基因。研究结果提示，这些分子或可为阐明肿瘤转移机制提供关键依据，亦有望成为肿瘤治疗的潜在分子靶标。整体实验设计：本研究通过对1例口腔鳞状细胞癌患者的肿瘤组织进行原代培养，成功建立原发灶来源的下颌牙龈癌细胞株WK2与转移灶来源的WK3F；随后采用基因芯片技术对二者的基因表达变化进行了全面分析。