Department of Otolaryngology, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung, Taiwan ; Kaohsiung Chang Gung Head and Neck Oncology Group, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan.
Department of Medical Research and Development, Show Chwan Memorial Hospital and Chang Bing Show Chwan Memorial Hospital, Changhua, Taiwan.
PLoS One. 2013 Dec 27;8(12):e84218. doi: 10.1371/journal.pone.0084218. eCollection 2013.
Nasopharyngeal carcinoma (NPC) is known for its high metastatic potential and locoregional recurrence, although the molecular alterations that are driving NPC metastasis remain unclear at this time. This study aimed to examine the expression of fibulin-5 in NPC, correlate the results with clinicopathological variables and survival, and to investigate the role of fibulin-5 in human NPC cell lines.
Standard semi-quantitative-RT-PCR, quantitative-RT-PCR, immunoblotting, and immunohistochemistry were used to investigate the mRNA and protein expression profiles of fibulin-5 in normal and NPC tissues. Immunohistochemistry of fibulin-5 was correlated with clinicopathological characteristics by univariate analyses. NPC cells overexpressing fibulin-5 or fibulin-5-siRNA cells were generated by stable transfection to characterize the molecular mechanisms of fibulin-5-elicited cell growth and metastasis.
Our results demonstrated that fibulin-5 overexpression in NPC specimens and significantly correlated with advanced tumor metastasis indicating a poor 5-year overall survival. Fibulin-5 was mainly expressed in the nucleus in human NPC specimens and cell lines. Functionally, fibulin-5 overexpression yielded fast growth in NPC cells. In addition, fibulin-5 promotes cell metastasis in NPC cells through increased FLJ10540 and phosphor-AKT activity. In contrast, siRNA depletion of fibulin-5 suppressed FLJ10540 expression and phosphor-AKT activity. Suppression of either fibulin-5 or FLJ10540 can cause significant inhibition with regards to cell motility in NPC cells. Finally, immunohistochemical analysis of human aggressive NPC specimens showed a significant and positive correlation between fibulin-5 and FLJ10540 expression.
Higher fibulin-5 expression is not only an important indicator of poor survival, but also contributes to the development of new therapeutic strategies in the FLJ10540/AKT pathway for NPC treatment.
鼻咽癌（NPC）以其高转移潜能和局部复发率而闻名，尽管目前尚不清楚驱动 NPC 转移的分子改变。本研究旨在研究纤维结合蛋白 5 在 NPC 中的表达，将结果与临床病理变量和生存相关联，并研究纤维结合蛋白 5 在人 NPC 细胞系中的作用。
采用标准半定量 RT-PCR、定量 RT-PCR、免疫印迹和免疫组织化学方法研究纤维结合蛋白 5 在正常和 NPC 组织中的 mRNA 和蛋白表达谱。通过单因素分析，免疫组织化学纤维结合蛋白 5 与临床病理特征相关联。通过稳定转染过表达纤维结合蛋白 5 或纤维结合蛋白 5-siRNA 细胞，以表征纤维结合蛋白 5 引发的细胞生长和转移的分子机制。
我们的结果表明，NPC 标本中纤维结合蛋白 5 的过度表达与晚期肿瘤转移显着相关，表明 5 年总生存率较差。纤维结合蛋白 5 主要在人 NPC 标本和细胞系中表达在细胞核中。功能上，纤维结合蛋白 5 的过度表达导致 NPC 细胞快速生长。此外，纤维结合蛋白 5 通过增加 FLJ10540 和磷酸化 AKT 活性促进 NPC 细胞的转移。相反，纤维结合蛋白 5 的 siRNA 耗尽抑制了 FLJ10540 的表达和磷酸化 AKT 活性。在 NPC 细胞中抑制纤维结合蛋白 5 或 FLJ10540 均可导致细胞迁移显著抑制。最后，对人侵袭性 NPC 标本的免疫组织化学分析显示，纤维结合蛋白 5 与 FLJ10540 表达之间存在显著正相关。
更高的纤维结合蛋白 5 表达不仅是生存不良的重要指标，而且还为 NPC 治疗中 FLJ10540/AKT 通路的新治疗策略的发展做出了贡献。
