Characterization of arecoline-induced effects on cytotoxicity in normal human gingival fibroblasts by global gene expression profiling.
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.
Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Taiwan, ROC.
Shang-Lun
SL
Chiang
Shih-Sheng
SS
Jiang
Yi-Jou
YJ
Wang
Horn-Che
HC
Chiang
Ping-Ho
PH
Chen
Hung-Pin
HP
Tu
Kun-Yen
KY
Ho
Yu-Shan
YS
Tsai
I-Shou
IS
Chang
Ying-Chin
YC
Ko
D001115:Arecoline / Q000633:toxicity*; D002460:Cell Line; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects* / Q000473:pathology; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects*; D005881:Gingiva / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D011619:Psychotropic Drugs / Q000633:toxicity*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014617:Vacuoles / Q000187:drug effects
D011619:Psychotropic Drugs; D001115:Arecoline