重组人核心蛋白聚糖基因增强多柔比星对白血病 K562细胞抑制作用的实验研究

 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-β1 mRNA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-β1 mRNA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-β1 mRNA的转录发挥其作用，其具体机制有待进一步研究。

Enhancement of recombinant human decorin gene to suppression effect of durorubicin on human leukemic K562 cell line

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.