ABSTRACT
Squamous cell carcinomas of the head and neck are characterized by their high tendency for invasion and metastasis. Several studies have identified the roles of matrix metalloproteinases (MMPs), vascular endothelial growth factors (VEGF) and urokinase plasminogen activators (uPA) in this process. Photodynamic Therapy (PDT) is an emerging treatment currently in clinical practice for the treatment of early cancer. Here we evaluate, in vitro, the influence of PDT on the expression of these molecules. A series of human keratinocyte cell lines derived from human oral squamous cell carcinomas (OSCC) were used as the PDT 'targets' in this study. Each cell line was subjected to sublethal dose of PDT. Activity of MMP-2, MMP-9, MMP-13, uPA and VEGF were evaluated at protein levels using zymography and ELISA on culture medium. For uPA, a chromogenic assay was performed. Gelatin zymography results revealed that, in control medium, MMP-9 and MMP-2 were secreted in proform. MMP-2 was highly expressed by H376 cells while VB6 and UP cells relatively show similar MMP-2 with comparatively low expression. For MMP-9, the latent type was highly expressed by VB6 cells and only slightly by H376, while active-MMP-9 was expressed by VB6 cell line only. Following PDT, both active and latent MMP-2 and MMP-9 were down regulated by UP and VB6 cells (p<0.001), while H376 showed an increase in active-MMP-2. These observations were supported by ELISA. This study has demonstrated that, PDT causes the suppression of factors responsible for tumour invasion which may be of therapeutic value.
Subject(s)
Carcinoma, Squamous Cell/drug therapy , Mouth Neoplasms/drug therapy , Photochemotherapy/methods , Antineoplastic Agents/therapeutic use , Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/physiopathology , Cell Line, Tumor , Cells, Cultured , Down-Regulation/drug effects , Down-Regulation/physiology , Humans , Keratinocytes/drug effects , Keratinocytes/physiology , Matrix Metalloproteinase 13/metabolism , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Mesoporphyrins/therapeutic use , Mouth Neoplasms/metabolism , Mouth Neoplasms/physiopathology , Neoplasm Invasiveness/physiopathology , Urokinase-Type Plasminogen Activator/metabolism , Vascular Endothelial Growth Factor A/metabolismABSTRACT
BACKGROUND: Early detection of premalignant/malignant lesions in the oral cavity can certainly improve the patient's prognosis. This study presents fluorescence imaging with the topical application of 5-aminolevulinic as a way to improve detection of various oral tissue pathologies. This procedure depends mainly on comparing the intensity of red and green fluorescence emitted from tissues during examination. MATERIALS AND METHODS: Seventy-one patients who presented with clinically suspicious oral leukoplakia were recruited for this study. Each of the patients was required to have 5-aminolevulinic acid in the form of mouth rinse prior to fluorescence imaging. Following this a surgical biopsy was acquired from the exact examination site. The results of the fluorescence spectroscopy have been compared with histopathology. RESULTS: A Student's t-test was applied to test the viability of the ratio between red and green fluorescence. The red-to-green ratio was found to increase significantly when the lesion was identified as dysplastic or carcinoma in situ. By applying a threshold line to discriminate between normal and dysplastic lesions; a sensitivity of 83-90% and specificity of 79-89% were obtained. CONCLUSION: Fluorescence spectroscopy combined with 5-aminolevulinic acid-induced protoporphyrin IX was found as a valuable tool in the diagnosis of oral premalignancy. This technique offers the potential to be advantageous over other non-optical techniques in terms of providing real-time diagnosis, in situ monitoring, cost effectiveness and more tolerated by patient compared to surgical biopsy.
