Over-expression of nerve growth factor-β in human cholangiocarcinoma QBC939 cells promote tumor progression

• Published in: PloS one 2013-04, Vol.8 (4), p.e62024
• Main Authors: Yue, Xiu-Jing, Xu, Lei-Bo, Zhu, Man-Sheng, Zhang, Rui, Liu, Chao
• Format: Article
• Language: English
• Subjects: Angiogenesis; Animals; Apoptosis; Apoptosis - genetics; Assaying; Biology; Biotechnology; Cell culture; Cell cycle; Cell Cycle - genetics; Cell growth; Cell Line, Tumor; Cell migration; Cell Movement - genetics; Cell Proliferation; Cell Transformation, Neoplastic - genetics; Cholangiocarcinoma; Cholangiocarcinoma - genetics; Cholangiocarcinoma - pathology; Colonies; Disease Progression; Endothelial cells; Female; Gene Expression; Gene Expression Regulation, Neoplastic; Gene Order; Gene Silencing; Heterografts; Humans; Infiltration; Liver cancer; Lymph nodes; Lymphatic system; Medical prognosis; Medicine; Metastases; Metastasis; Mice; Nerve growth factor; Nerve Growth Factor - genetics; Overexpression; Plasmids; Prostate; Rodents; Surgery; Tumor Burden - genetics; Tumorigenicity; Tumors; Vascular endothelial growth factor; Vascular endothelial growth factor C; Vascular Endothelial Growth Factor C - genetics
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0062024

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.