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目的:探讨小白菊内酯(parthenolide,PTL)对CD34+CD38-KG-1a白血病细胞的增殖、Bcl-2表达及其被同种异体NK(allo-NK)细胞杀伤敏感性的影响。方法:免疫磁珠法从KG-1a细胞中分离CD34+CD38-KG-1a细胞。XTT法检测PTL对CD34+CD38-白血病细胞增殖的影响,Real-time PCR和Western blotting分别检测PTL对CD34+CD38-KG-1a细胞Bcl-2mRNA和蛋白表达的影响,LDH释放法观察PTL对CD34+CD38-KG-1a细胞被allo-NK细胞杀伤敏感性的影响。结果:PTL对CD34+CD38-KG-1a细胞增殖的抑制呈剂量(2.5~80μmol/L)依赖性。PTL浓度为2.5μmol/L时,PTL组CD34+CD38-KG-1a细胞的增殖抑制率显著高于对照组[(4.89±1.07)%vs 0,P<0.01];PTL杀伤CD34+CD38-KG-1a细胞的IC50为20μmol/L;PTL组CD34+CD38-KG-1a细胞Bcl-2 mRNA[(0.105±0.007)vs(0.307±0.013),P<0.01]与蛋白表达均显著低于对照组。在效靶比为10∶1,20∶1和40∶1时,allo-NK细胞对PTL组CD34+CD38-KG-1a细胞杀伤率逐步升高,且均高于阴性(无PTL处理)对照组[(19.76±1.01)%,(30.14±0.96)%和(51.48±3.15)%vs(12.50±1.42)%,(16.90±0.93)%和(31.70±1.53)%(均P<0.01)];效靶比为40∶1时,allo-NK细胞对PTL组细胞的杀伤效率显著高于阳性(Bcl-2抑制剂ABT-737处理)对照组[(51.48±3.15)%vs(43.08±2.81)%,P<0.05]。结论:PTL能抑制CD34+CD38-KG-1a细胞的增殖并增强其被allo-NK细胞杀伤的敏感性,这可能与PTL下调Bcl-2的表达有关。
Objective: To investigate the effects of parthenolide (PTL) on the proliferation, the expression of Bcl-2 and the cytotoxicity of allo-NK cells in CD34 + CD38-KG-1a leukemia cells. Methods: CD34 + CD38-KG-1a cells were isolated from KG-1a cells by immunomagnetic beads method. The effect of PTL on the proliferation of CD34 + CD38-leukemia cells was detected by XTT method. The effect of PTL on the expression of Bcl-2 mRNA and protein in CD34 + CD38-KG-1a cells was detected by Real-time PCR and Western blotting respectively. + CD38-KG-1a cells by allo-NK cell killing sensitivity. Results: The inhibitory effect of PTL on the proliferation of CD34 + CD38-KG-1a cells was dose-dependent (2.5 ~ 80μmol / L). The proliferation inhibition rate of PTL group CD34 + CD38-KG-1a cells was significantly higher than that of the control group at PTL concentration of 2.5μmol / L [(4.89 ± 1.07)% vs 0, P <0.01] (P <0.01) and the protein expression of Bcl-2 mRNA in PTL group [(0.105 ± 0.007) vs (0.105 ± 0.007) vs 0.307 ± 0.013] were significantly lower than those in control group . The killing rates of allo-NK cells on CD34 + CD38-KG-1a cells in PTL group were gradually increased at the ratios of 10: 1, 20:1 and 40:1, and were higher than those of negative (no PTL treated) (19.76 ± 1.01)%, (30.14 ± 0.96)% and (51.48 ± 3.15)% vs (12.50 ± 1.42)%, (16.90 ± 0.93)% and (31.70 ± 1.53)%, respectively (51.48 ± 3.15)% vs (43.08 ± 2.81) in allo-NK cells in the PTL group was significantly higher than that in the control group (51.48 ± 3.15% vs 43.08 ± 2.81, the Bcl-2 inhibitor ABT- )%, P <0.05]. CONCLUSION: PTL can inhibit the proliferation of CD34 + CD38-KG-1a cells and increase its sensitivity to allo-NK cells, which may be related to the down-regulation of Bcl-2 expression by PTL.