Department of Hepato-Pancreato-Biliary Surgery, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Guangdong Province, China.
PLoS One. 2013 Apr 24;8(4):e62024. doi: 10.1371/journal.pone.0062024. Print 2013.
It has been shown that nerve growth factor-β (NGF-β) promoted the initiation and progression of many tumors, and we have previously demonstrated that the expression of NGF-β was associated with tumor stage, nerve infiltration and lymph node metastasis in human hilar cholangiocarcinoma. However, whether NGF-β promotes tumor progression in human cholangiocarcinoma requires further investigation. Therefore, we aimed to determine the effects of NGF-β on the progression of human cholangiocarcinoma.
Human cholangiocarcinoma QBC939 stable cell lines with over-expressed or silenced NGF-β genes were generated with pEGFP-N1-NGF-β and pGPU6/GFP/Neo-NGF-β-shRNA recombinant plasmids. Cell proliferation assay, colony formation assay, cell cycle analysis, apoptosis assay and tumorigenicity assay were performed to evaluate the role of NGF-β in the progression of human cholangiocarcinoma. In addition, human lymphatic endothelial cells were co-cultured with QBC939 culture supernatants, and the cell proliferation and migration abilities of the lymphatic endothelial cells were evaluated.
Forced expression of NGF-β in QBC939 cell lines promoted proliferation, colony formation and tumorigenicity in these cells and inhibited the apoptosis. However, down-regulation of NGF-β inhibited proliferation, colony formation and tumorigenicity, and increased the apoptotic rate of QBC939 cells. In addition, the NGF-β gain-of-function induced a high expression of vascular endothelial growth factor C and enhanced the proliferation and migration of lymphatic endothelial cells, while NGF-β loss-of-function showed opposite effects.
We concluded that NGF-β promoted tumor progression in human cholangiocarcinoma QBC939 cells. Our results provided a new concept to understand the role of NGF-β in cholangiocarcinoma progression, and might provide important information for the development of new targeted therapies in human cholangiocarcinoma.
已有研究表明神经生长因子-β（NGF-β）可促进多种肿瘤的发生和发展，我们之前的研究表明，NGF-β的表达与人类肝门部胆管癌的肿瘤分期、神经浸润和淋巴结转移有关。然而，NGF-β是否促进人类胆管癌的肿瘤进展仍需要进一步研究。因此，我们旨在确定 NGF-β对人类胆管癌细胞进展的影响。
通过 pEGFP-N1-NGF-β 和 pGPU6/GFP/Neo-NGF-β-shRNA 重组质粒，构建 NGF-β 过表达和基因沉默的人胆管癌细胞系 QBC939。通过细胞增殖实验、集落形成实验、细胞周期分析、细胞凋亡实验和肿瘤生成实验，评估 NGF-β在人类胆管癌进展中的作用。此外，将人淋巴管内皮细胞与 QBC939 培养上清液共培养，评估淋巴管内皮细胞的增殖和迁移能力。
在 QBC939 细胞系中强制表达 NGF-β可促进细胞增殖、集落形成和肿瘤生成，并抑制细胞凋亡。相反，下调 NGF-β 抑制了细胞增殖、集落形成和肿瘤生成，并增加了 QBC939 细胞的凋亡率。此外，NGF-β 功能获得诱导血管内皮生长因子 C 的高表达，并增强了淋巴管内皮细胞的增殖和迁移，而 NGF-β 功能丧失则表现出相反的效果。
我们得出结论，NGF-β 促进了人胆管癌细胞 QBC939 的肿瘤进展。我们的研究结果提供了一个新的概念，即了解 NGF-β 在胆管癌进展中的作用，并可能为人类胆管癌新的靶向治疗提供重要信息。
