摘要
背景与目的:在研究大蒜素抑制人胃癌细胞BGC823恶性增殖,将其阻滞于G1期并诱导BGC823细胞凋亡的分子机制及其靶基因过程中,我们发现cyclinD1和p27Kip1蛋白在G1期阻滞中起重要作用。本实验从mRNA和蛋白水平探讨大蒜素对人胃癌细胞BGC823cyclinD1和p27Kip1表达水平的影响。方法:用25μg/ml大蒜素处理BGC823细胞,并分别提取对照组和大蒜素处理组的总RNA和总蛋白,然后采用RT-PCR和Westernblot法检测大蒜素处理前后cyclinD1和p27Kip1mRNA和蛋白的变化。结果:经25μg/ml大蒜素处理BGC823细胞24h,cyclinD1蛋白表达下调,p27Kip1蛋白表达上调,处理48h上述变化更明显。而mRNA水平未见明显改变。结论:大蒜素可影响cyclinD1和p27Kip1蛋白的表达,而不影响mRNA的转录。
BACKGROUND &OBJECTIVE: Previous study showed that allitridi mar ke dly suppresses cellular proliferation, blocks cell cycle at G1 phase, and induce s cell apoptosis of human gastric cancer BGC823 cell. During the investigation o f its molecular mechanisms and target genes, the authors found that protein expr ession of cyclin D1 and p27Kip1 play an important role in an allitridi-induced G1 arrest. This study was designed to explore the effect of allitridi on the exp ression of cyclin D1 and p27Kip1 in BGC823 cells. METHODS: After total RNA and t otal protein in allitridi-treated (25 μg/ml) and allitridi-untreated BGC823 c ells were abstained, Western blot analysis was performed to determine the protei n levels of cyclin D1 and p27Kip1, and RT-PCR was performed to determine the mR NA levels of cyclin D1 and p27Kip1. RESULTS: After treated with 25 μg/ml allitr idi for 24 hours, the protein levels of cyclin D1 of BGC823 cells were downregul ated and the protein levels of p27Kip1 were upregulated, and this changes were m ore obvious after 48 hours,while their mRNA levels remained unchanged.CONCLUSION :Allitridi may influence the protein expression of cyclin D1 and p27Kip1 in BGC8 23 cells, while do not influence their mRNA transcription.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2003年第12期1268-1271,共4页
Chinese Journal of Cancer
基金
国家重点基础研究发展规划项目(No.98031200)
国家自然科学基金(No.39625016)
