CAJ | 학술논문

目的观察姜黄素对人胆囊癌细胞株GBC-SD的抗增殖效应,并探讨其作用机制.方法采用噻唑蓝(MTT)法检测0、10、30、50、70 mg/L浓度的姜黄素对胆囊癌GBC-SD细胞24 h的生长抑制作用；检测30 mg/L浓度的姜黄素在0、12、24、48、72 h对GBC-SD细胞的抑制作用.依照MTT结果设立对照组、10 mg/L姜黄素组、30 mg/L姜黄素组及50 mg/L姜黄素组分别作用GBC-SD细胞24 h.运用流式细胞术检测细胞的凋亡率,使用Western blot法和逆转录-聚合酶链反应(RT-PCR)法分别检测姜黄素对核因子(NF)-κB与细胞周期素(Cyclin) D1表达的影响,通过实时定量聚合酶链反应(Real-time PCR)法检测姜黄素对bax、B淋巴细胞/白血病-2(bcl-2) mRNA表达的影响.结果姜黄素以时间-剂量的方式抑制胆囊癌GBC-SD细胞的活力.10、30、50 mg/L浓度的姜黄素作用24 h后,细胞存活率分别为(95.34±9.21)％、(70.63±11.09)％、(50.58±6.31)％,与对照组比较,30、50 mg/L姜黄素组的差异有统计学意义(P＜0.05)；上述各组细胞的凋亡率分别为(9.83±1.98)％、(37.25±1.21)％和(57.63±7.38)％,与对照组比较,30、50 mg/L姜黄素组的差异有统计学意义(P＜0.05).30、50 mg/L的姜黄素均可下调NF-κB和Cyclin D1的表达,同时能够降低bcl-2、升高bax的表达,而10 mg/L姜黄素对上述基因表达无明显影响.结论姜黄素不仪能够通过下调NF-κB与Cyclin D1的表达,还能降低bel-2的mRNA水平、升高bax的mRNA水平导致bcl-2/bax的比值下降,从而有效地抑制胆囊癌GBC-SD细胞的增殖、诱导细胞凋亡.
목적 관찰강황소대인담낭암세포주GBC-SD적항증식효응,병탐토기작용궤제.방법 채용새서람(MTT)법검측0、10、30、50、70 mg/L농도적강황소대담낭암GBC-SD세포24 h적생장억제작용；검측30 mg/L농도적강황소재0、12、24、48、72 h대GBC-SD세포적억제작용.의조MTT결과설립대조조、10 mg/L강황소조、30 mg/L강황소조급50 mg/L강황소조분별작용GBC-SD세포24 h.운용류식세포술검측세포적조망솔,사용Western blot법화역전록-취합매련반응(RT-PCR)법분별검측강황소대핵인자(NF)-κB여세포주기소(Cyclin) D1표체적영향,통과실시정량취합매련반응(Real-time PCR)법검측강황소대bax、B림파세포/백혈병-2(bcl-2) mRNA표체적영향.결과 강황소이시간-제량적방식억제담낭암GBC-SD세포적활력.10、30、50 mg/L농도적강황소작용24 h후,세포존활솔분별위(95.34±9.21)％、(70.63±11.09)％、(50.58±6.31)％,여대조조비교,30、50 mg/L강황소조적차이유통계학의의(P＜0.05)；상술각조세포적조망솔분별위(9.83±1.98)％、(37.25±1.21)％화(57.63±7.38)％,여대조조비교,30、50 mg/L강황소조적차이유통계학의의(P＜0.05).30、50 mg/L적강황소균가하조NF-κB화Cyclin D1적표체,동시능구강저bcl-2、승고bax적표체,이10 mg/L강황소대상술기인표체무명현영향.결론 강황소불의능구통과하조NF-κB여Cyclin D1적표체,환능강저bel-2적mRNA수평、승고bax적mRNA수평도치bcl-2/bax적비치하강,종이유효지억제담낭암GBC-SD세포적증식、유도세포조망.
Objective To explore the anti-proliferation effect of curcumin on human gallbladder carcinoma GBC-SD cells and its mechanism.Methods The growth inhibition of GBC-SD cells treated with curcumin of 0,10,30,50,and 70 mg/L was detected by using methyl thiazol tetrazolium (MTT) assay.The growth inhibition of GBC-SD cells treated with curcumin of 30 mg/L at 0,12,24,48 and 72 h was detected.Control group,10 mg/L curcumin group,30 mg/L curcumin group and 50 mg/L curcumin group were set up.The apoptosis rate was tested by using flow cytometry.After curcumin treatment for 24 h,the expression levels of nuclear factor-κB (NF-κB) and Cyclin D1 were detected by using Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR),respectively.The mRNA levels of bax and B lymphocytes/leukemia-2 (bcl-2) were examined by using real-time reverse transcription polymerase chain reaction.Results Curcumin could significantly inhibit the viability of GBC-SD cells in a time-and dosedependent manner.After treatment with curcumin (10,30,and 50 mg/L),the survival rate was (95.34 ±9.21)％,(70.63 ± 11.09)％ and (50.58 ± 6.31)％,respectively.There was significant difference between control group and 30 mg/L curcumin group or 50 mg/L curcumin group (P ＜ 0.05) ;The apoptosis rate was (9.83 ± 1.98)％,(37.25 ± 1.21)％ and (57.63 ±7.38)％,respectively.There was significant difference between control group and 30 mg/L curcumin group or 50 mg/L curcumin group (P ＜ 0.05).30 mg/L and 50 mg/L curcumin could down-regulate the expression of NF-κB and CyclinD1,reduce the expression of bcl-2 and enhance the expression of bax.However,10 mg/L curcumin had no evident effect on genes mentioned above.Conclusion Curcumin could not only down-regulate the expression of NF-κB and Cyclin D1,but also reduce the mRNA level of bcl-2 and up-regulate the mRNA level of bax,causing the reduced bcl-2/bax ratio,which could effectively inhibit the proliferation of gallbladder carcinoma GBC-SD cells and induce cell apoptosis.