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目的 探讨吲哚美辛 (IN)抗慢性髓系白血病 (CML)细胞增殖的作用机制。方法 用MTT法分析IN对慢性髓系白血病细胞系K5 6 2及CML患者骨髓细胞活力的影响 ;Westernblot分析CML细胞信号转导和转录活化蛋白 (STAT) 1、STAT5表达 ;免疫共沉淀 /Westernblot分析IN干预后CML细胞磷酸化的STAT1(p STAT1)和STAT5 (p STAT5 )表达 ,间接免疫荧光技术观察p STAT1和p STAT5在CML细胞中的分布及变化 ;Westernblot检测IN干预前后Bcl XL 及环氧化酶 2 (COX 2 )蛋白的表达。结果 IN可显著抑制CML细胞活力 ;4 0 0 μmol/LIN作用 72h ,CML细胞抑制率为 86 % ;0～ 4 0 0 μmol/LIN呈剂量反应性抑制p STAT1、p STAT5表达 ;p STAT主要呈散点状分布于胞核中 ;Bcl XL 蛋白呈IN浓度依赖性表达下调 ;K5 6 2细胞存在COX 2蛋白表达 ,IN可抑制COX 2蛋白表达。结论 IN可明显抑制CML细胞增殖 ;其机制可能与药物抑制STAT/Bcl XL 信号转导途径有关 ;K5 6 2细胞存在COX 2蛋白表达 ,IN的抗白血病细胞增殖效应可能是通过COX 2依赖性途径实现的。
Objective To investigate the mechanism of indomethacin (IN) against chronic myeloid leukemia (CML) cell proliferation. Methods MTT assay was used to analyze the effect of IN on the myeloid cell viability in chronic myeloid leukemia cell line K562 and CML. Western blotting was used to analyze the signal transducers and activators of transcription 1 (STAT1) and STAT5 in CML cells. Immunoprecipitation / Western blot analysis The expression of STAT1 (STAT1) and STAT5 (STAT5) of phosphorylated STAT3 (p STAT5) and STAT5 (p STAT5) in CML cells after IN intervention were detected by indirect immunofluorescence. The expressions of p STAT1 and p STAT5 in CML cells were detected by Western blot. 2 (COX 2) protein expression. RESULTS IN inhibited the viability of CML cells, the inhibitory rate of CML cells was 86% after treated with 400 μmol / L for 72 hours, and the levels of p STAT1 and p STAT5 were dose-dependently decreased from 0 to 400 μmol / Bcl XL was down-regulated in IN concentration-dependent manner; COX 2 protein was found in K5 6 2 cells and IN inhibited COX 2 protein expression. CONCLUSION: IN can significantly inhibit the proliferation of CML cells. The mechanism may be related to the inhibition of STAT / Bcl XL signal transduction pathway. The expression of COX 2 protein in K562 cells may be related to the inhibitory effect of IN on leukemia cell proliferation through COX 2-dependent pathway Achieve.