CAJ | 학술논문

目的探讨C-myc与β-Catenin介导的肺腺癌A549/DDP的耐药相关性。方法 MTT细胞毒实验比较A549与A549/DDP细胞对顺铂的敏感性；Western blot检测在顺铂作用前、后β-catenin及C-myc在肺腺癌A549及A549/DDP中的蛋白表达及免疫荧光定位；流式细胞仪检测不同剂量的顺铂作用下两株细胞以及A549/DDP转染C-myc siRNA前后的凋亡和周期变化。结果 MTT细胞毒实验结果显示顺铂对A549和A549/DDP细胞的IC50值分别为（5.769±0.24）μmol/L和（28.373±0.96）μmol/L，与A549细胞比较． A549/DDP顺铂耐药4.92倍。 Western blot 结果显示β-Catenin 及C-myc蛋白在A549/DDP细胞的表达较A549高，顺铂作用48 h后A549细胞的β-catenin及C-myc表达较顺铂作用前下调（ P＝0.007）， A549/DDP细胞的β-catenin及C-myc表达较顺铂作用前上调（ P＝0.006）；免疫荧光定位结果显示顺铂作用48 h后A549及A549/DDP细胞的β-Catenin由膜/质/核表达转为质/核表达，C-myc表达由浆/核表达转为核表达。流式结果显示随着顺铂剂量的增加，细胞凋亡率成剂量依赖性增加。 A549/DDP中转染C-myc siRNA 48 h后引起细胞S期阻滞，G0/G1期细胞积聚，凋亡率增加（P＝0.013），IC50值明显下降（P＝0.009）。结论β-catenin的下游靶基因C-myc在调节A549/DDP的耐药性中起到一定作用，C-myc siRNA可以提高A549/DDP细胞对顺铂的敏感性。
목적탐토C-myc여β-Catenin개도적폐선암A549/DDP적내약상관성。방법 MTT세포독실험비교A549여A549/DDP세포대순박적민감성；Western blot검측재순박작용전、후β-catenin급C-myc재폐선암A549급A549/DDP중적단백표체급면역형광정위；류식세포의검측불동제량적순박작용하량주세포이급A549/DDP전염C-myc siRNA전후적조망화주기변화。결과 MTT세포독실험결과현시순박대A549화A549/DDP세포적IC50치분별위（5.769±0.24）μmol/L화（28.373±0.96）μmol/L，여A549세포비교． A549/DDP순박내약4.92배。 Western blot 결과현시β-Catenin 급C-myc단백재A549/DDP세포적표체교A549고，순박작용48 h후A549세포적β-catenin급C-myc표체교순박작용전하조（ P＝0.007）， A549/DDP세포적β-catenin급C-myc표체교순박작용전상조（ P＝0.006）；면역형광정위결과현시순박작용48 h후A549급A549/DDP세포적β-Catenin유막/질/핵표체전위질/핵표체，C-myc표체유장/핵표체전위핵표체。류식결과현시수착순박제량적증가，세포조망솔성제량의뢰성증가。 A549/DDP중전염C-myc siRNA 48 h후인기세포S기조체，G0/G1기세포적취，조망솔증가（P＝0.013），IC50치명현하강（P＝0.009）。결론β-catenin적하유파기인C-myc재조절A549/DDP적내약성중기도일정작용，C-myc siRNA가이제고A549/DDP세포대순박적민감성。
Objective To investigate the C-myc and β-catenin mediated drug resistance in lung adenocarcinoma cell A549/DDP.Methods Cisplatin sensitivity was determined by MTT cell toxicity experiment in A549 and A549/DDP cells.Western blot and immunofluorescence methods were used to determine the expression ofβ-catenin and C-myc protein after being treated with cisplatin in A 549 and A549/DDP cells.Flow cytometry was performed to detect cell cycle and apoptosis in A 549,A549/DDP and C-myc siRNA transfected A549/DDP before and after adding different doses of cisplatin .Results MTT cytotoxic experiment results showed that IC 50 in A549 and A549/DDP cells treated with cisplatin was (5.769 ±0.24 )μmol/L and(28.373 ±0.96 )μmol/L respectively, and A549 cells cisplatin resistance was 4.92 times of A549/DDP.Western blot showed that both endogenous β-catenin and C-myc expression in A549/DDP cells were higher than that in A549,and were upregulated in A549/DDP (P=0.006)downregulated in A549 after 48 h treatment with cisplatin(P=0.007);Immunofluorescence showed that the expression of β-catenin was transferred from membrane/cytoplasma/nuclear to cytoplasma/nuclear, and the expression of C-myc was transferred from cytoplasma/nuclear to nuclear in both cells after being treated with cisplatin.Flow cytometry demonstrated that with the increase of dose cisplatin ,cell apoptosis rate increased in dose dependent manner .After 48 h transfection with C-myc siRNA in the A549/DDP,cells were blocked in S phase and G0/G1 phase cells were increased .Simultaneously , cell apoptosis rate were increased ( P=0.013 ) and the IC50 decreased significantly ( P =0.009 ).Conclusion C-myc, the downstream target genes of β-catenin, plays an important role in regulating A 549/DDP cisplatin resistance .C-myc siRNA could improve the sensitivity of A 549/DDP cells to cisplatin.