HSP90靶向抑制剂P7与阿霉素联用对急性髓性白血病细胞凋亡的协同增效作用
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摘要
目的探讨热休克蛋白HSP90靶向抑制多肽LPLTPLP(P7)与临床标准化疗药物阿霉素(DOX)联用对急性髓性白血病细胞系NB4的影响。方法多肽P7与DOX单药或两药联合处理NB4细胞系48 h后,CCK-8法检测细胞的存活,并用等效图解法解析其作用;流式细胞计量术检测NB4细胞凋亡水平;蛋白免疫印迹检测NB4细胞凋亡蛋白PARP的表达水平及剪切水平。结果 P7与DOX对细胞存活具有协同抑制作用;P7与DOX联合用药组细胞凋亡率为63.9%,约等于P7和DOX两药单独用药时凋亡率之和;两药联用组凋亡相关蛋白PARP总蛋白表达显著下调(P<0.01),cleaved-PARP的水平显著上调(P<0.01)。结论 P7协同DOX能显著提高NB4细胞的凋亡比例,其机制可能与DNA修复酶PARP的失活有关。
Objective To explore the effect of doxorubicin(DOX) combined with a heat shock protein 90 targeted heptapeptide-LPLTPLP(P7) in the acute myeloid leukemia cell line NB4 in vitro. Methods Following treatment with P7, DOX, or P7 combined with DOX in NB4 cells for 48 h, cell viability was assessed by CCK-8 assay and analyzed with the isobologram method;flow cytometry was carried out to quantify apoptotic cells;the expression and cleavage level of PARP protein were tested by Western blot analysis. Results P7 synergized with DOX in inhibiting cell viability. The apoptotic rate of combination of P7 and DOX was approximately equal to the sum of P7 and DOX apoptotic rate. The protein expression of PARP was significantly decreased while that of cleaved PARP was significantly increased with the treatment of P7 or/and DOX. Conclusions Combination of P7 and DOX exhibits synergistic effects on NB4 cells by increasing apoptosis rate, the mechanism of which may be associated with the inactivation of DNA repair enzyme PARP.
Objective To explore the effect of doxorubicin(DOX) combined with a heat shock protein 90 targeted heptapeptide-LPLTPLP(P7) in the acute myeloid leukemia cell line NB4 in vitro. Methods Following treatment with P7, DOX, or P7 combined with DOX in NB4 cells for 48 h, cell viability was assessed by CCK-8 assay and analyzed with the isobologram method;flow cytometry was carried out to quantify apoptotic cells;the expression and cleavage level of PARP protein were tested by Western blot analysis. Results P7 synergized with DOX in inhibiting cell viability. The apoptotic rate of combination of P7 and DOX was approximately equal to the sum of P7 and DOX apoptotic rate. The protein expression of PARP was significantly decreased while that of cleaved PARP was significantly increased with the treatment of P7 or/and DOX. Conclusions Combination of P7 and DOX exhibits synergistic effects on NB4 cells by increasing apoptosis rate, the mechanism of which may be associated with the inactivation of DNA repair enzyme PARP.
引文
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[7] Georgakis GV,Li Y,Rassidakis GZ,et al.The HSP90 inhibitor 17-AAG synergizes with doxorubicin and U0126 in anaplastic large cell lymphoma irrespective of ALK expression[J].Exp Hematol,2006,34:1670-1679.
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[12] Bergeron C,Thiesse P,Rey A,et al.Revisiting the role of doxorubicin in the treatment of rhabdomyosarcoma:an up-front window study in newly diagnosed children with high-risk metastatic disease[J].Eur J Cancer,2008,44:427-431.
[13] Yang X,Wang J.Precision therapy for acute myeloid leukemia[J].J Hematol Oncol,2018,11:3-13.
[14] Lim SH,Dubielecka PM,Raghunathan VM.Molecular targeting in acute myeloid leukemia[J].J Transl Med,2017,15:183-195.