目的:探讨蛋白激酶C(PKC)抑制剂米哚妥林(PKC412)对HL-60细胞株增殖和凋亡的影响及作用机制。方法:采用CCK-8法检测不同浓度PKC412对HL-60细胞增殖的影响;瑞士吉姆萨染色法鉴定PKC412对HL-60细胞凋亡的影响;qRT-PCR检测BCL-2、P53基因的...目的:探讨蛋白激酶C(PKC)抑制剂米哚妥林(PKC412)对HL-60细胞株增殖和凋亡的影响及作用机制。方法:采用CCK-8法检测不同浓度PKC412对HL-60细胞增殖的影响;瑞士吉姆萨染色法鉴定PKC412对HL-60细胞凋亡的影响;qRT-PCR检测BCL-2、P53基因的mRNA表达,Western blot检测BCL-2、P53蛋白的表达;以尾静脉注射HL-60细胞构建急性白血病小鼠模型,以PKC41231.25nmol/kg尾静脉给药,生理盐水组作为对照组,研究PKC412在小鼠体内对HL-60细胞的抑制;ELISA法检测PKC412对肿瘤小鼠细胞因子TNF-α和TGF-β分泌的影响。结果:PKC412抑制HL-60细胞株细胞增殖,呈剂量依赖性(r=0.9973),其中IC50=0.31μmol/L;并能诱导HL-60细胞凋亡。PKC412作用HL-60细胞株48 h后,与对照组比较BCL-2基因mRNA表达下调(0.417±0.044 vs 0.933±0.033,t=9.347,P<0.001),P53基因mRNA表达上调(1.533±0.145 vs 1.050±0.161,t=2.231,P>0.05);BCL-2蛋白表达量下降,P53蛋白表达量升高。PKC412在体内可抑制HL-60肿瘤细胞生长,给药后小鼠生存率为50%,与对照组比较体重增加(18.02±0.403 g vs 16.44±0.562 g,t=2.272,P=0.0356)。PKC412组血清及脾细胞中TNF-α和TGF-β细胞因子较对照组分泌明显减少(P<0.05)。结论:PKC412能通过抑制BCL-2基因表达水平诱导HL-60细胞凋亡,体内给药可抑制肿瘤的生长。展开更多
Background: Patients with stem cell myeloproliferative disorders have a particularly poor prognosis and limited treatment options, i.e. mainly aggressive chemotherapy or allogeneic stem cell transplantation. In 2004, ...Background: Patients with stem cell myeloproliferative disorders have a particularly poor prognosis and limited treatment options, i.e. mainly aggressive chemotherapy or allogeneic stem cell transplantation. In 2004, Chen et al. reported a patient presenting a t(8;13) (p11;q12) cytogenic anomaly who responded positively to treatment with PKC412 (midostaurin), an oral multi-targeted tyrosine kinase inhibitor. Here, we report a second case treated with the above-mentioned drug. Patient: A 71-year-old woman was diagnosed as having chronic myelogenous leukaemia with eosinophilia secondary to t(8;13) with FGFR1 involvement. Due to her age, an allogeneic stem cell transplantation was not possible. Treatment: A treatment combining aggressive chemotherapy and midostaurin was explored. The patient received one cycle of hyper-CVAD chemotherapy followed by maintenance therapy with midostaurin. A relapse occurred after six months, and she was treated with four more cycles of hyper-CVAD chemotherapy. The patient entered a complete clinical, haematological and cytogenetic remission. A maintenance therapy with midostaurin continued for four months until she developed a chemoresistant relapse followed by acute leukaemia. Conclusion: This is the second case of a t(8;13) myeloid and lymphoid neoplasm with FGFR1 abnormalities treated successfully with midostaurin. Midostaurin is administered orally, allows for outpatient care and in this case showed only occasional and minimal side effects. The combination of hyper-CVAD and midostaurin extended survival by 21 months without allogeneic transplantation. This case further supports the possibility of using midostaurin for the treatment of other diseases with FGFR1 dysregulations;however, specific clinical trials are needed to confirm this hypothesis.展开更多
文摘目的:探讨蛋白激酶C(PKC)抑制剂米哚妥林(PKC412)对HL-60细胞株增殖和凋亡的影响及作用机制。方法:采用CCK-8法检测不同浓度PKC412对HL-60细胞增殖的影响;瑞士吉姆萨染色法鉴定PKC412对HL-60细胞凋亡的影响;qRT-PCR检测BCL-2、P53基因的mRNA表达,Western blot检测BCL-2、P53蛋白的表达;以尾静脉注射HL-60细胞构建急性白血病小鼠模型,以PKC41231.25nmol/kg尾静脉给药,生理盐水组作为对照组,研究PKC412在小鼠体内对HL-60细胞的抑制;ELISA法检测PKC412对肿瘤小鼠细胞因子TNF-α和TGF-β分泌的影响。结果:PKC412抑制HL-60细胞株细胞增殖,呈剂量依赖性(r=0.9973),其中IC50=0.31μmol/L;并能诱导HL-60细胞凋亡。PKC412作用HL-60细胞株48 h后,与对照组比较BCL-2基因mRNA表达下调(0.417±0.044 vs 0.933±0.033,t=9.347,P<0.001),P53基因mRNA表达上调(1.533±0.145 vs 1.050±0.161,t=2.231,P>0.05);BCL-2蛋白表达量下降,P53蛋白表达量升高。PKC412在体内可抑制HL-60肿瘤细胞生长,给药后小鼠生存率为50%,与对照组比较体重增加(18.02±0.403 g vs 16.44±0.562 g,t=2.272,P=0.0356)。PKC412组血清及脾细胞中TNF-α和TGF-β细胞因子较对照组分泌明显减少(P<0.05)。结论:PKC412能通过抑制BCL-2基因表达水平诱导HL-60细胞凋亡,体内给药可抑制肿瘤的生长。
文摘Background: Patients with stem cell myeloproliferative disorders have a particularly poor prognosis and limited treatment options, i.e. mainly aggressive chemotherapy or allogeneic stem cell transplantation. In 2004, Chen et al. reported a patient presenting a t(8;13) (p11;q12) cytogenic anomaly who responded positively to treatment with PKC412 (midostaurin), an oral multi-targeted tyrosine kinase inhibitor. Here, we report a second case treated with the above-mentioned drug. Patient: A 71-year-old woman was diagnosed as having chronic myelogenous leukaemia with eosinophilia secondary to t(8;13) with FGFR1 involvement. Due to her age, an allogeneic stem cell transplantation was not possible. Treatment: A treatment combining aggressive chemotherapy and midostaurin was explored. The patient received one cycle of hyper-CVAD chemotherapy followed by maintenance therapy with midostaurin. A relapse occurred after six months, and she was treated with four more cycles of hyper-CVAD chemotherapy. The patient entered a complete clinical, haematological and cytogenetic remission. A maintenance therapy with midostaurin continued for four months until she developed a chemoresistant relapse followed by acute leukaemia. Conclusion: This is the second case of a t(8;13) myeloid and lymphoid neoplasm with FGFR1 abnormalities treated successfully with midostaurin. Midostaurin is administered orally, allows for outpatient care and in this case showed only occasional and minimal side effects. The combination of hyper-CVAD and midostaurin extended survival by 21 months without allogeneic transplantation. This case further supports the possibility of using midostaurin for the treatment of other diseases with FGFR1 dysregulations;however, specific clinical trials are needed to confirm this hypothesis.