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Isolation, Characterization and Functional Examination of the Gingival Immune Cell Network
Published on: February 16, 2016
Characterization of arecoline-induced effects on cytotoxicity in normal human gingival fibroblasts by global gene
Shang-Lun Chiang1, Shih-Sheng Jiang, Yi-Jou Wang
1Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Taiwan, ROC.
Abstract:
Areca nut is the most widely used psychoactive substance and an important environmental risk factor for development of oral premalignant lesions and cancer. Arecoline, the major alkaloid of areca nut, has been known to cause cytotoxicity and genotoxicity in mammalian cells in vivo and in vitro and even contributes to carcinogenicity. However, the susceptible genes accounting for arecoline-induced damage in normal human oral cells are still lacking, which possibly involves in initial molecular damage via alternation of gene expression level on biological pathways. The present study was undertaken to characterize the toxic effects of arecoline in gene expression profiling on normal human gingival fibroblasts (HGF) using cDNA microarray and quantitative real-time reverse transcription PCR. The cytotoxicity of arecoline on HGF-1 cell line was elevated in a dose-dependent manner (p < 0.05) accompanied with distinct morphological change and formation of intracellular vacuoles were observed. At optimum concentration of arecoline determined from dose-response curve of the cytotoxicity, a large number of genes were significantly repressed than induced by arecoline in global gene expression profiling. Five induced- and seven repressed genes including glutathione synthetase were further validated, and their gene expression changes were increased in a dose-dependent manner in a concentration range of 50-150 microg/ml. In conclusion, we proposed a tentative model to explain arecoline-induced effects on contribution of oral pathogenesis. The findings identified that 12 susceptible genes can potentially serve as biomarkers of arecoline-induced damage in betel chewers.
Insights
Arecoline, found in areca nut, damages human oral cells by altering gene expression. This study identifies 12 genes that may serve as biomarkers for arecoline-induced oral damage in betel quid consumers.
Area of Science:
- Oral pathology
- Molecular toxicology
- Genomics
Background:
- Areca nut is a widely consumed psychoactive substance linked to oral cancer.
- Arecoline, its main alkaloid, exhibits cytotoxicity and genotoxicity, but susceptible genes in oral cells are unknown.
- Understanding gene expression changes is crucial for identifying early molecular damage pathways.
Purpose of the Study:
- To investigate the toxic effects of arecoline on gene expression in normal human gingival fibroblasts (HGF).
- To identify specific genes affected by arecoline exposure.
- To propose potential biomarkers for arecoline-induced oral pathogenesis.
Main Methods:
- Utilized cDNA microarray and quantitative real-time reverse transcription PCR to analyze gene expression.
- Assessed arecoline's cytotoxicity and morphological effects on HGF-1 cells in a dose-dependent manner.
- Validated expression levels of selected genes, including glutathione synthetase.
Main Results:
- Arecoline exposure increased cytotoxicity and caused cellular vacuolation in HGF-1 cells.
- Global gene expression profiling revealed more repressed than induced genes following arecoline treatment.
- Expression of 12 genes, including glutathione synthetase, was significantly altered in a dose-dependent manner.
Conclusions:
- Identified 12 genes susceptible to arecoline-induced damage in normal human oral cells.
- These genes show potential as biomarkers for detecting arecoline-induced damage in betel quid users.
- A model for arecoline's role in oral pathogenesis was proposed based on gene expression alterations.