CAJ | 학술논문

目的探讨重组人核心蛋白聚糖(rhDCN)基因增强多柔比星(ADM)对人类白血病K562细胞的作用.方法取对数生长期的K562细胞分为0.9%NaCl溶液组、pcDNA3.1(+)-DCN/K562组、ADM/K562组、pcDNA3.1(+)-DCN加ADM/K562组.瑞特染色观察细胞形态改变,MTT法检测细胞增殖活性,流式细胞术(FCM)分析细胞凋亡,RT-PCR分析各组TGF-β_1mRNA含量.结果 pcDNA3.1(+)-DCN加ADM/K562组细胞比单独DCN和ADM组细胞,染色呈现更显著的凋亡形态学改变;MTT法结果显示联合组细胞增殖抑制率为(61±1.32)%,明显高于单独干预组[DCN组(20±1.90)%;ADM组(47±1.04)%](P<0.05);FCM检测结果显示联合组细胞凋亡指数为(61.30±0.9)%,较单独干预组[DCN组(28.25±1.3)%及ADM组(31.85±1.5)%]明显增加(P<0.05);RT-PCR结果显示,联合组细胞TGF-β_1mRNA的转录减少.结论 rhDCN可以明显增强ADM对K562细胞的杀伤作用,提高肿瘤细胞的凋亡率.rhDCN可能通过下调TGF-β_1mRNA的转录发挥其作用,其具体机制有待进一步研究.
목적 탐토중조인핵심단백취당(rhDCN)기인증강다유비성(ADM)대인류백혈병K562세포적작용.방법 취대수생장기적K562세포분위0.9%NaCl용액조、pcDNA3.1(+)-DCN/K562조、ADM/K562조、pcDNA3.1(+)-DCN가ADM/K562조.서특염색관찰세포형태개변,MTT법검측세포증식활성,류식세포술(FCM)분석세포조망,RT-PCR분석각조TGF-β_1mRNA함량.결과 pcDNA3.1(+)-DCN가ADM/K562조세포비단독DCN화ADM조세포,염색정현경현저적조망형태학개변;MTT법결과현시연합조세포증식억제솔위(61±1.32)%,명현고우단독간예조[DCN조(20±1.90)%;ADM조(47±1.04)%](P<0.05);FCM검측결과현시연합조세포조망지수위(61.30±0.9)%,교단독간예조[DCN조(28.25±1.3)%급ADM조(31.85±1.5)%]명현증가(P<0.05);RT-PCR결과현시,연합조세포TGF-β_1mRNA적전록감소.결론 rhDCN가이명현증강ADM대K562세포적살상작용,제고종류세포적조망솔.rhDCN가능통과하조TGF-β_1mRNA적전록발휘기작용,기구체궤제유대진일보연구.
Objective To investigate the suppression effect, the apoptosis and TGF-β_1 mRNA expression of rhDCN and dororubicin(ADM) on leukemic K562 cell line. Methods K562 cells in Logarithmic growth phase were divided into Saline group, pcDNA3.1 (+)-DCN group, ADM group, and pcDNA3.1 (+)-DCN-ADM group. Morphology change of cell was detected by Wright stain, cell proliferation activity was assessed by MTT. The apoptosis index of K562 cells was assessed by FCM, and TGF-β_1 mRNA of cell was assessed by RT-PCR. Results Wright stain showed that more pronounced morphological apoptosis changes of K562 cells in combined group. MTT method results showed that the proliferation inhibition rate of the combined group was (61±1.32) % higher than that of individual intervention group [DCN group, (20±1.90) %; ADM group, (47±1.04) %](P <0.05). FCM results showed that the apoptosis index of the combined group was (61.30± 0.9) %, higher than that of Individual intervention group [DCN group, (28.25±1.3) %; ADM group, (31.85± 1.5) %](P <0.05). TGF-β_1 mRNA synthesis of combined group was significantly decreased. Conclusion rhDCN can markedly enhance cytotoxicity of ADM on K562 cells, and the mechanisms of apoptosis may be due to down-regulation of TGF-β_1 mRNA. Specific mechanisms will be further studied.