IADR Abstract Archives
Identification of Novel Neuronal Roles in Oral Cancer Tumour Microenvironments
Objectives: Nerves have recently been implicated in breast, pancreatic and prostate cancer progression, but the molecular mechanisms underlying this are poorly understood. Little is known regarding the contribution of nerves to disease progression in oral cancer. This study's objective was to investigate neuronal-tumour interactions in oral squamous cell carcinomas (OSCCs).
Methods: Quantitative polymerase chain reaction (qPCR) and immunocytochemistry were used to identify the neuropeptides calcitonin gene-related peptide (CGRP), substance P (SP) and components of their receptors in normal oral keratinocytes (NOKs), dysplastic, cancerous and metastatic cell lines and neurones. Co-culture systems were used to identify functional cross-talk mediated by neuronal, cancer cell and fibroblast (the predominant cell type of the tumour microenvironment) secretions. ELISA and qPCR were employed to identify secreted factors playing roles in neural-tumour interactions.
Results: PCR identified CGRP and SP receptors in most OSCC-derived cell lines. Immunocytochemistry localised receptor components within OSCC and H357 cells. CGRP and SP were detected in primary neurones used for co-culture systems. Significant chemoattractant and proliferative effects of both neuropeptides were identified in H357 cells in a dose- and time-dependent manner. Nerve growth factor (NGF) was elevated in most cancer-derived cell lines relative to NOKs and its secretion detected from H357 cells. NGF secretion from experimentally derived cancer-associated fibroblasts was significantly increased compared to normal oral fibroblasts which correlated with increased neuronal outgrowth. H357 caused significant increases in neuronal outgrowth.
Conclusions: The results indicate the existence of significant cross-talk between nerves and the OSCC tumour microenvironment. This suggests neuronal factors may have potential as therapeutic targets.
Methods: Quantitative polymerase chain reaction (qPCR) and immunocytochemistry were used to identify the neuropeptides calcitonin gene-related peptide (CGRP), substance P (SP) and components of their receptors in normal oral keratinocytes (NOKs), dysplastic, cancerous and metastatic cell lines and neurones. Co-culture systems were used to identify functional cross-talk mediated by neuronal, cancer cell and fibroblast (the predominant cell type of the tumour microenvironment) secretions. ELISA and qPCR were employed to identify secreted factors playing roles in neural-tumour interactions.
Results: PCR identified CGRP and SP receptors in most OSCC-derived cell lines. Immunocytochemistry localised receptor components within OSCC and H357 cells. CGRP and SP were detected in primary neurones used for co-culture systems. Significant chemoattractant and proliferative effects of both neuropeptides were identified in H357 cells in a dose- and time-dependent manner. Nerve growth factor (NGF) was elevated in most cancer-derived cell lines relative to NOKs and its secretion detected from H357 cells. NGF secretion from experimentally derived cancer-associated fibroblasts was significantly increased compared to normal oral fibroblasts which correlated with increased neuronal outgrowth. H357 caused significant increases in neuronal outgrowth.
Conclusions: The results indicate the existence of significant cross-talk between nerves and the OSCC tumour microenvironment. This suggests neuronal factors may have potential as therapeutic targets.