CAJ | 학술논문

目的：研究水飞蓟宾联合5-氟尿嘧啶（5-FU ）抑制胃癌MGC-803细胞增殖，探讨其协同增效作用及机制．方法：采用噻唑蓝（MTT）法检测不同浓度的水飞蓟宾对人胃癌MGC-803细胞，人正常肝L02细胞的增殖抑制作用，并计算IC20，IC50；采用IC20浓度的水飞蓟宾联合不同浓度的5-氟尿嘧啶，观察对人胃癌MGC-803细胞增殖抑制作用；水飞蓟宾单独或联合5-FU作用于胃癌MGC-803细胞，碘化丙锭（PI）单染色检测细胞周期改变，Annexin V-FITC/PI双染流式细胞术检测细胞凋亡水平，Western blotting检测细胞周期和凋亡相关蛋白的表达．结果：MTT法检测显示，水飞蓟宾对胃癌MGC-803细胞有增殖抑制作用，且呈剂量-效应关系，对人正常肝L02细胞增殖抑制作用较弱．水飞蓟宾作用于胃癌MGC-803细胞48 h 的IC20、IC50浓度分别为144.6、214.7μmol /L；水飞蓟宾与不同浓度的5-氟尿嘧啶联合作用于MGC-803细胞48 h，可提高MGC-803细胞对5-氟尿嘧啶的敏感性，增敏5.73倍．流式结果显示，无论是单独使用水飞蓟宾、5-氟尿嘧啶还是水飞蓟宾与5-氟尿嘧啶联合使用都能使细胞抑制在G0/G1期和出现凋亡细胞群；Western blotting结果显示，水飞蓟宾与5-氟尿嘧啶联合使用，能使细胞的细胞周期相关蛋白P15表达升高，CDK6表达下降，Bcl-2蛋白家族中抗凋亡蛋白Bcl-2，Bcl-xL表达明显降低，促凋亡蛋白Bax表达基本不变，Casepase-9，Caspase-3活化降解．结论：水飞蓟宾提高胃癌MGC-803细胞对5-FU的敏感性，水飞蓟宾在联合5-FU之后能够使MGC-803细胞抑制在G0/G1期，并通过线粒体凋亡途径使细胞发生凋亡．
목적：연구수비계빈연합5-불뇨밀정（5-FU ）억제위암MGC-803세포증식，탐토기협동증효작용급궤제．방법：채용새서람（MTT）법검측불동농도적수비계빈대인위암MGC-803세포，인정상간L02세포적증식억제작용，병계산IC20，IC50；채용IC20농도적수비계빈연합불동농도적5-불뇨밀정，관찰대인위암MGC-803세포증식억제작용；수비계빈단독혹연합5-FU작용우위암MGC-803세포，전화병정（PI）단염색검측세포주기개변，Annexin V-FITC/PI쌍염류식세포술검측세포조망수평，Western blotting검측세포주기화조망상관단백적표체．결과：MTT법검측현시，수비계빈대위암MGC-803세포유증식억제작용，차정제량-효응관계，대인정상간L02세포증식억제작용교약．수비계빈작용우위암MGC-803세포48 h 적IC20、IC50농도분별위144.6、214.7μmol /L；수비계빈여불동농도적5-불뇨밀정연합작용우MGC-803세포48 h，가제고MGC-803세포대5-불뇨밀정적민감성，증민5.73배．류식결과현시，무론시단독사용수비계빈、5-불뇨밀정환시수비계빈여5-불뇨밀정연합사용도능사세포억제재G0/G1기화출현조망세포군；Western blotting결과현시，수비계빈여5-불뇨밀정연합사용，능사세포적세포주기상관단백P15표체승고，CDK6표체하강，Bcl-2단백가족중항조망단백Bcl-2，Bcl-xL표체명현강저，촉조망단백Bax표체기본불변，Casepase-9，Caspase-3활화강해．결론：수비계빈제고위암MGC-803세포대5-FU적민감성，수비계빈재연합5-FU지후능구사MGC-803세포억제재G0/G1기，병통과선립체조망도경사세포발생조망．
Aim:To investigate the synergistic effect of Silibinin combined with 5-fluorouracil (5-FU) on gastric cancer MGC-803 cells and the possible mechanism.Methods:MTT assay was used to deter-mine the inhibitory effect of Silibinin on gastric cancer MGC-803 cells and hepatocyte L02 cells.Changes in cell cycle and the apoptotic cell percentage were determined by Flow Cytometry.The expression of cell cycle related proteins and apoptosis associated proteins were analyzed by Western blotting.Results:MTT method was used to determine the IC20 value and IC50 value for hepatocyte L02 cells.IC20 concentra-tion of Silibinin combined with 5-FU at different concentrations were incubated with MGC-803 cells for 48 hours,MTT assay was used to measure the proliferation inhibition effects.After MGC-803 cells were in-cubated with Silibinin combined with 5-FU for 48 hours,the cell cycle of MGC-803 was inhibited by Sili-binin at different concentrations,in a dose-dependent manner(P<0.05 ),but no obvious effects were found on the proliferation of hepatocyte L02 cells.The IC20 and IC50 for Silibinin against gastric cancer MGC-803 cells after incubation for 48 hours were found to be 144.6 and 214.7 μmol /L,respectively. The chemo-therapeutic effects of 5-fluorouracil on MGC-803 cells were enhanced when combined with Sil-ibinin,and the drug sensitivity was increased by about 5 .73 times.PI staining analysis showed that cell cycle was arrested in G0/G1 phase.Annexin V-FITC /PI staining analysis also showed a dose-dependent effect on the apoptotic cells.Western blotting showed that the expression of cell cycle related protein CDK6 was decreased while that of P15 was increased.The expressions of apoptosis associated proteins, pro-Caspase-9,pro-Caspase-3,Bcl-2,Bcl-xL,were found to be decreased significantly,but expression of Bax remained unchanged.Conclusion:Silibinin has synergistic inhibitory effect with 5-fluorouracil on MGC-803 cells.Silibinin combined with 5-fluorouracil might inhibit cell cycle in G0/G1 phase and in-duce cell apoptosis through the mitochondrial pathway.