异硫氰酸苯己酯对K562／G01细胞伊马替尼耐药性的影响

Effect of phenylhexyl isothiocyanate on drug - resistance to imatinib in K562/G01 cell line

目的体外观察异硫氰酸苯己酯（PHI）对人慢性髓性白血病细胞株K562／G01细胞伊马替尼耐药性的影响，探讨其可能的机制。方法采用MTT法检测不同浓度的PHI和伊马替尼单独或联合处理对K562／G01细胞增殖的抑制作用。流式细胞术检测不同浓度的PHI和（或）伊马替尼作用下K562／G01细胞的凋亡率。Westernblot检测PHI处理K562／G01细胞后P—gP、P210^bcr-abl蛋白、磷酸化P210^bcr-abl蛋白（p-P210^bcr-abl）表达水平的变化。结果PHI单独处理24h可抑制K562／G01细胞增殖，诱导细胞凋亡。PHI浓度由0增至40Ixmol／L，细胞增殖抑制率由0增至（51．22-4-1．41）％，凋亡率由（3．764-1．46）％增至（35．354-3．70）％。浓度分别为10、20、40txmol／L的PHI与不同浓度伊马替尼联合处理后，伊马替尼的IC50分别为（10．494-1．24）、（6．334-1．42）、（0．854-0．17）μmoL／L。PHI20txmol／L和浓度为10、20Ixmol／L的伊马替尼分别共同作用24h后，K562／G01细胞凋亡率分别为（43．624-4．23）％和（55．414-4．35）％，较单用伊马替尼组及单用PHI组均明显升高。PHI浓度由0增至40txmol／L分别作用7h后，K562／G01细胞P210^bcr-abl／β-actin比值由0．9444-0．034降至0．392±0．025，p-P210^bcr-abl／β-actin比值由0．9064-0．019降至0．3614-0．021，但P—gP表达无明显变化。结论PHI能抑制K562／G01细胞增殖，诱导细胞凋亡。PHI与伊马替尼联合有协同作用，可部分逆转细胞对伊马替尼的耐药，其机制可能与抑制P210^bcr-abl和p-P210^bcr-abl蛋白表达有关。

Objective To investigate the effect of phenylhexyl isothiocyanate （PHI） on the drug-re- sistance to imatinib in K562/G01 cell line and to elucidate its possible mechanisms. Methods MTT assay was employed to access K562/GO1 cell growth inhibition after exposure to PHI and/or imatinib at different concentrations. Apoptotic rate of K562/G01 cells was measured by flow eytometry. The levels of P-gp, P210^bcr-abl and p-P210^bcr-abl protein were detected by Western blot. Results PHI inhibited proliferation and induced apoptosis of K562/GO1 cells after treated with PHI alone for 24 h. PHI concentration increased from 0 to 40 μmoL/L, the inhibitory rate of cell proliferation from 0 to （51.22 ± 1.41 ） % , and the apoptosis rate from （3.76± 1.46） % to （35.35 ± 3.70） %. Combination of 10, 20, 40 jxmol/L PHI and various concen- trations of imatinib, ICs0s of imatinib were （ 10.49 ± 1.24）, （6.33 ±1.42）, and （0.85 ± 0.17 ） μmol/L, respectively. When K562/G01 cells treated with 20 p, mol/L PHI combined with 10 and 20txmoL/L imatinib for 24 hours, apoptosis rate were （43.62 ± 4.23 ） % and （55.41 ± 4.35） % , respectively, being significant- ly higher than that with imatinib or PHI alone. PHI concentrations increased from 0 to 40 μmol/L for 7 hours, the P210^bcr-abl/β-actin decreased from （0.944±0. 034） to （0. 392 ± 0. 025 ） , and the p-P210^bcr-abl/β-actin decreased from （0. 906 ±0. 019） to （0. 361 ± 0. 021 ） , while the alteration of P-gp was not seen. Conclu- sions PHI inhibits the proliferation and induces apoptosis of K562/G01 cell line. PHI has synergistic effect with imatinib. It partially reverses the drug-resistance to imatinib. The mechanism of reversal of drug resist- ance in K562/G01 cells might be by inhibitinz P210^bcr-abl and o-P210^bcr-abl