CAJ | 학술논문

目的探讨全反式维甲酸(ATRA)诱导HL-60细胞分化模型中血管内皮生长因子(VEGF)表达调控的分子机制,为白血病的抗血管新生治疗提供新的靶点.方法 Wright-Gimesa染色观察细胞形态,硝基四唑氮蓝(NBT)还原实验检测HL-60细胞分化,逆转录-聚合酶链反应(RT-PCR)、蛋白质印记(Western blot)分别从mRNA和蛋白水平检测细胞VEGF、信号转导和转录激活因子-3(STAT3)、c-myc的表达变化.结果 1μmolATRA作用于HL-60细胞96 h后可明显抑制细胞增殖,并出现明显的分化现象,倒置显微镜下可观察到细胞向成熟粒细胞方向分化；NBT阳性率为82.59％(t=-24.157,P＜0.01)；VEGF mRNA表达水平下降(t=7.339,P＜0.05)、STAT3mRNA表达水平下降(t=3.667,P＜0.05)、c-myc mRNA表达水平下降(t=6.858,P＜0.05).VEGF蛋白表达水平下降(t=3.386,P＜0.05)、STAT3蛋白表达水平下降(t=4.074,P＜0.05)、c-myc蛋白表达水平下降(t=3.333,P ＜0.05).结论在ATRA诱导HL-60细胞分化的模型中VEGF的表达水平随诱导分化的进程而下降,其下调机制可能与STAT3、c-myc具有相关性.
목적 탐토전반식유갑산(ATRA)유도HL-60세포분화모형중혈관내피생장인자(VEGF)표체조공적분자궤제,위백혈병적항혈관신생치료제공신적파점.방법 Wright-Gimesa염색관찰세포형태,초기사서담람(NBT)환원실험검측HL-60세포분화,역전록-취합매련반응(RT-PCR)、단백질인기(Western blot)분별종mRNA화단백수평검측세포VEGF、신호전도화전록격활인자-3(STAT3)、c-myc적표체변화.결과 1μmolATRA작용우HL-60세포96 h후가명현억제세포증식,병출현명현적분화현상,도치현미경하가관찰도세포향성숙립세포방향분화；NBT양성솔위82.59％(t=-24.157,P＜0.01)；VEGF mRNA표체수평하강(t=7.339,P＜0.05)、STAT3mRNA표체수평하강(t=3.667,P＜0.05)、c-myc mRNA표체수평하강(t=6.858,P＜0.05).VEGF단백표체수평하강(t=3.386,P＜0.05)、STAT3단백표체수평하강(t=4.074,P＜0.05)、c-myc단백표체수평하강(t=3.333,P ＜0.05).결론 재ATRA유도HL-60세포분화적모형중VEGF적표체수평수유도분화적진정이하강,기하조궤제가능여STAT3、c-myc구유상관성.
Objective To study the molecular regulation mechanism of VEGF in the model of ATRA induced differentiation in HL-60 cells,and to provide new targets for leukemia anti-angiogenic therapy.Methods The morphology was observed by Wright-Gimesa staining; HL-60 cells differentiation was detected by NBT reduction experiment.VEGF,STAT3,c-myc mRNAs were measured by reverse transcription-PCR;VEGF,STAT3 and c-myc proteins were determined by Western blot.Results The proliferation of HL-60 cells was inhibited obviously by ATRA(1 μmol/L) with the induction of differentiation,NBT positive rate was 82.59％ (t =-24.157,P ＜ 0.01) ; VEGF mRNA (t =7.339,P ＜ 0.05),STAT3 mRNA (t =3.667,P ＜0.05) and c-myc mRNA (t =6.858,P ＜ 0.05) were all down-regulated.VEGF protein (t =3.386,P ＜0.05),STAT3 protein(t =4.074,P ＜ 0.05) and c-myc protein (t =3.333,P ＜ 0.05) were all down-regulated.Conclusion VEGF expression level is reduced with the procession of differentiation of HL-60 cells,which may be largely correlated with the down regulation of STAT3 and c-myc.