摘要
目的进一步明确熊果酸(ursolic acid,UA)抑制胃癌细胞环氧化酶-2(cycloxygenase-2,COX-2)表达的信号转导通路。方法人胃腺癌细胞株SGC-7901和MKN-45常规培养于RPMI-1640培养液中,细胞长至亚单层后分别加抗氧化剂N-乙酰-L半胱氨酸(NAC)、单磷酸腺苷激活的蛋白激酶(AMP-activated protein kinase,AMPK)激活剂5-氨基咪唑-4-甲酰胺核苷酸(AICAR)、AMPK抑制剂compound C和信号转导与转录活化因子3(signal transducer and activator of transcription 3,STAT3)抑制剂WP1066预处理后再加UA连续培养24 h,Western blot检测AMPK、STAT3磷酸化水平和COX-2蛋白表达。结果抗氧化剂NAC和AMPK抑制剂compound C有效地阻断了UA抑制STAT3磷酸化和COX-2表达的作用,AMPK激活剂AICAR抑制STAT3磷酸化和COX-2表达,AICAR和UA联合作用大于单用,STAT3抑制剂WP1066对UA诱导的AMPK磷酸化无明显影响,WP1066单独或联合UA均可抑制STAT3磷酸化和COX-2表达且联合作用大于单用。结论 UA通过ROS/AMPK/STAT3信号转导通路抑制胃癌细胞COX-2表达。
Aim Our previous study has found that ur-solic acid（ UA） increased intracellular reactive oxygen species （ ROS ） production and adenosine monophos-phate-activated protein kinase （ AMPK ） phosphoryla-tion, inhibited signal transducer and activator of tran-scription 3 （ STAT3 ） phosphorylation and cyclooxygen-ase-2 （ COX-2 ） expression in gastric cancer cells . However , the molecular mechanism by which UA in-hibits COX-2 expression in gastric cancer cells has not been fully clarified .In this study we aimed to further clarify the signal transduction pathways involved in the UA-mediated inhibition of COX-2 expression in gastric＆nbsp;cancer cells .Methods Human gastric cancer cell lines SGC-7901 and MKN-45 were routinely cultured in RPMI-1640 medium supplemented with 10% heat-in-activated fetal calf serum .Sub-confluent cell cultures were pre-treated with antioxidant N-acetylcysteine （ NAC） , AMPK activator 5-amino-4-imida-zolecarbox-amide-riboside （ AICAR ） , AMPK inhibitor compound C, or STAT3 inhibitor WP1066 and then treated with or without UA for 24 h.The expression of AMPK and phosphorylated AMPK （ p-AMPK ） , STAT3 and phos-phorylated STAT3 （ p-STAT3 ） , as well as COX-2 was detected by Western blot analysis .Results Antioxi-dant NAC and AMPK inhibitor compound C blocked UA-induced inhibition of STAT 3 phosphorylation and down-regulation of COX-2 expression in gastric cancer cells.Both AMPK activator AICAR and UA inhibited＆nbsp;STAT3 phosphorylation and COX-2 expression; the combination of two drugs resulted in further reduction . STAT3 inhibitor WP1066 did not affect UA-induced AMPK phosphorylation , whereas it inhibited STAT3 phosphorylation and COX-2 expression .The inhibitory effects on the STAT3 phosphorylation and COX-2 ex-pression were significantly enhanced when SGC-7901 and MKN-45 cells were treated simultaneously with WP1066 plus UA.Conclusion UA inhibits COX-2 expression in gastric cancer cells , which may be medi-ated through ROS/AMPK/STAT3 signal transductio
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第7期925-932,共8页
Chinese Pharmacological Bulletin
基金
国家自然科学基金面上项目(No 81372659)
关键词
熊果酸
胃癌
活性氧
单磷酸腺苷激活的蛋白激酶
信号转导与转录活化因子3
环氧化酶-2
ursolic acid
gastric cancer
reactive oxy-gen species
adenosine monophosphate-activated pro-tein kinase
signal transducer and activator of tran-scription 3
cyclooxygenase-2
