反义VEGFcDNA转染联合应用IFNα或信号转导抑制剂571(STI571)对K562细胞的协同抑制

Synergistic inhibitory effect of antisense vascular endothelial growth factor cDNA transfection and IFNα or signal transduction inhibitor 571 (STI571) on the K562 cells

目的 :探讨转染反义VEGFcDNA及联合应用IFNα或STI571对慢性髓性白血病 (chronicmyeloidleukemia ,CML)急变细胞株K562的作用。方法 :利用转染反义 (AS)、正义 (S)VEGF1 2 1 cDNA及空载体 (V ,pcDNA3)的K562细胞株 ,采用台盼蓝拒染法、流式细胞术Annexin V FITC/PI双标技术 ,观察IFNα或STI571(CGP571 4 8B)对转染后K562细胞增殖及凋亡的影响。结果 :IFNα(1 0 0u·ml 1 )能抑制K562 /AS细胞生长 ,增加其凋亡。STI571 (0 .1 μmol·L 1 )对K562 /V、K562 /S和K562 /AS细胞生长均有抑制作用。低剂量VP1 6(1 0mg·L 1 )或低剂量STI571 (0 .1 μmol·L 1 )在 2 4h就对K562 /AS细胞有诱导凋亡的作用 ,且VP1 6(1 0mg·L 1 )与STI571(0 .1 μmol·L 1 )的联合应用具有协同诱导K562 /AS细胞凋亡的作用。在 72h时 ,0 .1 μmol·L 1 的STI571对K562 /V、K562 /S和K562 /AS细胞均有诱导凋亡的作用。结论 :转染反义VEGF1 2 1 cDNA联合应用IFNα能协同抑制细胞增殖 ,转染反义VEGF1 2 1 cDNA能增加K562细胞对VP1 6或STI571的敏感性。提示抗VEGF药物可能成为治疗CML的新策略。

Objective: To investigate the co operative effect of antisense VEGF 121 cDNA transfection and IFNα or STI571 on the chronic myeloid leukemia(CML) cell line K562. Methods: K562 cells which were transfected with the VEGF 121 cDNA antisense (AS) or sense (S) vector, and with the vector (V, pcDNA3) alone were utilized. Effects of IFNα or STI571 on the proliferation and apoptosis of transfected K562 cells were studied by Trypan blue stain,flow cytometric Annexin V FITC/PI dual labeling technique. Results: Antisense VEGF 121 cDNA transfection in combination with IFNα 100 u·ml 1 , 1 000 u·ml 1 and 10 000 u·ml 1 could inhibit the proliferation, increase the apoptosis. STI571 (0.1 μmol·L 1 ) could inhibit the proliferation of all the transfected cells. VP16(10 mg·L 1 )or /and STI571(0.1 μmol·L 1 ) could induce more apoptosis in K562/AS cells after 24 h. STI571 (0.1 μmol·L 1 ) could induce the apoptosis of all the transfected cells after 72 h. Conclusion: Antisense VEGF 121 cDNA transfected K562 cells show that they can be synergetically inhibited cell growth with IFNα,and that they are more susceptible to low dose VP16 or /and STI571. This suggests anti VEGF medicine may be a new therapy strategy against CML.