全反式维甲酸对K562细胞的诱导分化作用

Differentiation Inducing Effect of ATRA on Leukemia Cell Line K562

摘要:

目的：全反式维甲酸(ATRA)对肿瘤细胞具有广泛的抑制增殖、促进分化作用。本研究通过体外培养检测ATRA对K562细胞是否具有诱导分化作用。方法：采用联苯胺染色、瑞氏染色、非特异性酯酶染色、硝基四唑氮蓝还原试验4种不同的染色方法及流式细胞术,观察经1μmol/L及2.5μmol/L ATRA诱导1d,4d,5d后,K562细胞出现诱导分化特征。结果：1μmol/L及2.5μmol/L ATRA均可诱导K562细胞向粒系分化。培养4d,1μmol/L ATRA组61.5％的K562细胞、2.5μmol/L组39％的K562细胞显示出向粒系成熟方向分化;培养5d,处理组的分化细胞数仍明显高于对照组(P<0.05),但两种浓度ATRA的诱导分化强度无统计学差异(P>0.05)。且均未出现向红系或单核细胞方向分化的特征。流式细胞仪检测ATRA诱导前后CD13及CD71的表达,1μmol/L ATRA诱导K562细胞1d,CD13抗原表达阳性率为8.0％,2.5μmol/L组则为6.7％,均高于对照组(2.1％)。培养5d,1μmol/L和2.5μmol/L AFRA组的CD13分别升高到28.1％,37.8％,而CD71则分别下降至1.2％和0.9％。与对照组比差异均具有显著性(P<0.05)。结论：ATRA可诱导K562细胞向粒系成熟方向分化。

Abstract:

OBJECTIVE: ATRA can restrain proliferation and promote differentiation in various tumor cells. The aim of the study is to investigate the differentiation characteristics of K562 induced by ATRA. METHODS: Morphology (Benzidine staining, Wright's staining, NSE staining and NBT recovery test) and flow cytometry were used to observe the differentiation characteristics of K562 after co-incubation with 1 μmol/L and 2.5 pmol/L ATRA for 1 d, 4 ds, and 5 ds. RESULTS: Co-incubated for 4 ds, 61.5% K562 cells in the 1 μmol/L ATRA group and 39% K562 cells in the 2.5 μmol/L ATRA group showed some evidence of myeloid maturation, but no evidence of erythroid or monocytoid maturation. Cb-incubated for 5 ds, the percentage of differentiated K562 cells was much higher than that in the control group. One day after induction by 1 μmol/L ATRA or 2.5 μmol/L ATRA, the expression of CD13 was 8.0% and 6.7%, respectively, which was higher than that in the control group (2.1%). Five days after induction by 1 μmol/L ATRA or 2.5 μmol/L ATRA, the expression of CD13 increased to 28.1 % and 37.8 % , respectively, while the expression of CD71 decreased to 1.2 % and 0.9 % respectively. The differences between the ATRA groups and the control group were significant ( P < 0.05). CD71 decreased from 9.7% and 10.8% in the 1 μmol/L and 2.5 μmol/L ATRA groups on day 1 to 1.2% and 0.9% on day 5, while the CD13 expression level increased from 8.0% and 6.7% to 28.1% and 37.8% , respectively. CONCLUSIONS: ATRA can induce K562 to differentiate into myeloid linage. [Chin J Contemp Pediatr, 2003, 5(1): 8-11]