槟榔碱诱导上皮细胞cyclinD1表达下调

Arecoline Down-regulated the Expression of cyclinD1 in Hacat Cells

目的:探讨槟榔碱抑制上皮细胞增殖的分子机制。方法:应用光学显微镜、RT-PCR、报告基因、Western blot、免疫沉淀等方法,研究槟榔碱对细胞增殖的影响及分子机理。结果:槟榔碱处理上皮细胞Hacat 24h后,细胞增殖抑制,cyclinD1的转录抑制、启动子活性下调、蛋白表达降低及泛素化降解上调。结论:cyclinD1转录抑制和蛋白降解是槟榔碱抑制上皮细胞增殖的重要分子机制之一。

Objective： To investigate the mechanism of arecoline mediated epithelial cell proliferation inhibition. Methods： Light microscope, the reverse transcription--polymerase chain reaction technique, Western blot and re- porter gene assay, CO--immunopreeipitation were performed to study the effects of arecoline on cell proliferation and the underlying molecular mechanisms. Results： After treated with various concentrations of arecoline for 24 hours, the number of proliferating cells was obviously decreased and the gene transcription as well as promoter ac- tivity of cyclinD1 were dramatically declined. Arecoline dose-- dependently inhibits the protein expression of cy- clinD1 by up--regulation ubiquitination mediated eyclinD1 degradation. Conclusion： Transcription inhibition and protein degradation of eyclinD1 might play an important role in arecoline induced epithelial cell proliferation inhibi- tion.