CAJ | 학술논문

目的观察姜黄素对体外培养HepG2中p连环蛋白(p catenin)信号通路的影响,探讨姜黄素抑制肝母细胞瘤的分子机制.方法实时定量聚合酶链反应(RT PCR)及免疫印迹法(Western Blot)检测对照组及不同浓度姜黄素干预组(10、20、30、40、50μmol/L) HepG2中p-catenin mRNA及蛋白表达水平;TOP flash/FOP flash双荧光素酶报告系统检测β-catenin的转录活性;RTPCR检测各组-catenin靶基因细胞周期素D1(cyclin D1)、血管内皮细胞生长因子(VEGF)及基质金属蛋白酶9 (MMP9) mRNA表达水平.结果 HepG2高表达β-catenin mRNA及蛋白,各浓度姜黄素干预均能抑制HepG2细胞β-catenin mRNA表达(P＜0.05或P＜0.01),10～50μmol/L姜黄素干预分别降低了ycatenin mRNA表达水平20％、40％、48％、58％和60％;20～50μmol/L姜黄素干预均能抑制cyclin D1、VEGF及MMP9 mRNA的表达(P＜0.05或P＜0.01);20～50μmol/L姜黄素干预均能抑制HepG2细胞β-catenin蛋白表达(P＜0.05或P＜0.01),分别降低30％、43％、60％和75％;20～50μmol/L姜黄素干预分别抑制HepG2中pcatenin的转录活性27％、36％、55％和60％ (P＜0.05或P＜0.01).结论姜黄素可能通过抑制-catenin的表达及转录活性而影响β-catenin信号通路相关下游靶基因的表达,这可能是其发挥抗肿瘤效应的分子机制之一.
목적 관찰강황소대체외배양HepG2중p련배단백(p catenin)신호통로적영향,탐토강황소억제간모세포류적분자궤제.방법 실시정량취합매련반응(RT PCR)급면역인적법(Western Blot)검측대조조급불동농도강황소간예조(10、20、30、40、50μmol/L) HepG2중p-catenin mRNA급단백표체수평;TOP flash/FOP flash쌍형광소매보고계통검측β-catenin적전록활성;RTPCR검측각조-catenin파기인세포주기소D1(cyclin D1)、혈관내피세포생장인자(VEGF)급기질금속단백매9 (MMP9) mRNA표체수평.결과 HepG2고표체β-catenin mRNA급단백,각농도강황소간예균능억제HepG2세포β-catenin mRNA표체(P＜0.05혹P＜0.01),10～50μmol/L강황소간예분별강저료ycatenin mRNA표체수평20％、40％、48％、58％화60％;20～50μmol/L강황소간예균능억제cyclin D1、VEGF급MMP9 mRNA적표체(P＜0.05혹P＜0.01);20～50μmol/L강황소간예균능억제HepG2세포β-catenin단백표체(P＜0.05혹P＜0.01),분별강저30％、43％、60％화75％;20～50μmol/L강황소간예분별억제HepG2중pcatenin적전록활성27％、36％、55％화60％ (P＜0.05혹P＜0.01).결론 강황소가능통과억제-catenin적표체급전록활성이영향β-catenin신호통로상관하유파기인적표체,저가능시기발휘항종류효응적분자궤제지일.
Objective To explore the effects of curcumin (cur) on β-catenin signaling pathways in HepG2 and elucidate its anti tumor molecular mechanism.Methods The expressions of mRNA and protein of β-catenin in control HepG2 and curcumin treatment groups of different concentrations (10,20,30,40,50amol/L) were evaluated by reverse transcription polymerase chain reaction (RT PCR)and Western blot.And the transcriptional activity of β catenin was assayed by TOP flash/FOP flash dual luciferase reporter system.The expressions of β-catenin targeted gene cyclin D1,VEGF and MMP9 were evaluated by RT PCR.Results β-catenin was highly expressed in HepG2.Curcumin (10 50amol/L) could inhibit the mRNA expression of β-catenin by approximately 20％,40％,48％,58％ and 60％ respectively.Meanwhile curcumin (20 50amol/L) significantly inhibited the mRNA expression of β-catenin targeted gene MMP9,cyclin D1 and VEGF (P＜0.05 or P＜0.01).The expression of β-catenin at the protein level decreased by approximately 30％,43％,60％ and 75％after treatment.And the transcriptional activity of β-catenin in HepG2 was also inhibited by different concentrations of curcumin (20 50 μmol/L) and decreased by approximately 27％,36％,55％ and 60％ respectively (P＜0.05 or P＜0.01).Conclusions Curcumin can inhibit the expression and transcriptional activity of β-catenin and affect the expression of β-catenin targeted gene through the β catenin signaling pathways.And it may be a possible mechanism of its anti neoplastic effect.