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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。
Objective All-trans retinoic acid (ATRA) on tumor cells with a wide range of inhibition of proliferation, and promote differentiation. In this study, we examined whether ATRA induces differentiation of K562 cells in vitro. Methods Four different staining methods, such as benzidine staining, Wright’s staining, nonspecific esterase staining and nitroblue tetrazolium blue-reduction test, and flow cytometry were used to observe the effect of ATRA induced by 1μmol / L and 2.5μmol / L ATRA for 1 day, After 4d and 5d, K562 cells showed differentiation characteristics. Results Both Aμmol / L and 2.5μmol / L ATRA could induce K562 cells to differentiate into granulocytes. After cultured for 4 days, 61.5% of K562 cells in 1μmol / L ATRA group and 39% of K562 cells in 2.5μmol / L group showed differentiation toward mature lines. After cultured for 5 days, the number of differentiated cells in treated group was still significantly higher than that in control group (P < 0.05). However, there was no significant difference in the intensity of differentiation between the two concentrations of ATRA (P> 0.05). And did not appear to erythroid or monocyte differentiation characteristics. The expression of CD13 and CD71 before and after ATRA induction was detected by flow cytometry. The positive rate of CD13 antigen expression in K562 cells treated with 1μmol / L ATRA for 1 day was 8.0% and that in 2.5μmol / L group was 6.7% ). After cultured for 5 days, CD13 in 1μmol / L and 2.5μmol / L AFRA groups increased to 28.1% and 37.8%, respectively, while CD71 decreased to 1.2% and 0.9% respectively. Compared with the control group, the difference was significant (P <0.05). Conclusion ATRA can induce K562 cells to differentiate into mature lines.