Uptake and intracellular distribution of idarubicin in secondary cultures of normal and neoplastic urothelium

This study analyzes the uptake and endocellular distribution of idarubicin (IDA) in normal and neoplastic urothelial secondary cultures in relation to the changes in concentration and time of exposure. The urothelial lines were isolated by Freshney's method from biopsy fragments taken from five patients with superficial bladder cancer. Pharmacological experiments were carried out on subcultures previously immunophenotypically characterized and did not exceed ten passages. The uptake and endocellular distribution of IDA was analyzed by densitometric image analysis on cells treated for 10, 20, 30 and 60 min and 2 h with scalar dosages from 10 ng/ml to 2430 ng/ml. Microscopic observations and densitometric analyzes revealed that in the cells treated with IDA, fluorescence was higher in the cytoplasm compared to the nucleus and increased with the change in dosage. Moreover, densitometric data showed that IDA uptake in the first 20 min was higher in the neoplastic cells, but after that period its behavior became heterogeneous at 30 and 60 min, while at 2 h there was an inversion of the trend. These results suggest that the in vitro cytotoxicity should be evaluated in order to verify whether the elevated uptake of IDA in the first 20 min of treatment is really correlated to a more elevated toxicity in the neoplastic cells with respect to the normal cells. This is presently under investigation.     

联合去甲氧柔红霉素治疗急性白血病的临床研究

采用以４－去甲氧基柔红霉素（ＩＤＡ）为主组成的联合化疗方案，治疗３３例初发和复发的急性白血病，其中急性淋巴细胞白血病（ＡＬＬ）７例，急性非淋巴细胞白血病（ＡＮＬＬ）２６例。结果：总有效率７０％。初治２３例ＡＮＬＬ患者，完全缓解（ＣＲ）１６例，部分缓解（ＰＲ）２例，有效率为７９％。５例初治ＡＬＬ患者，４例ＣＲ，１例ＰＲ。而复发的２例ＡＬＬ和３例ＡＮＬＬ患者均未缓解。ＩＤＡ主要副作用表现为骨髓抑制及心脏毒性。认为以ＩＤＡ组成联合化疗方案治疗初发的急性白血病具有较好的疗效     

雷帕霉素对去甲氧柔红霉素诱导急性髓系白血病THP-1细胞凋亡的影响

目的:探讨雷帕霉素（Rapa）对去甲氧柔红霉素（IDA）诱导急性髓系白血病THP-1细胞凋亡的影响及其分子机制。方法:分别用10、20、40、80 nmol/L Rapa处理THP-1细胞1 h，另设未经Rapa处理的细胞。采用蛋白质印迹法检测THP-1细胞自噬标志物LC3蛋白的转换情况（LC3Ⅱ/LC3Ⅰ），采用流式细胞术检测细胞凋亡，确定Rapa处理浓度。用不同浓度IDA作用THP-1细胞24 h，采用CCK-8法检测IDA对THP-1细胞的增殖抑制率，计算半数抑制浓度（ IC50）。以低于 IC50的IDA作用Rapa处理或未处理的THP-1细胞24 h，CCK-8法检测细胞增殖抑制率，流式细胞术检测细胞凋亡情况，实时荧光定量聚合酶链反应检测自噬相关基因Beclin-1、LC3和p62的表达变化，蛋白质印迹法检测自噬标志物LC3蛋白的转换情况。 结果:20 nmol/L Rapa处理的THP-1细胞LC3Ⅱ/LC3Ⅰ高于未处理的细胞（ P=0.002 4）；80 nmol/L Rapa处理的细胞凋亡率高于未处理的细胞（ P=0.007 3）。根据蛋白质印迹法和流式细胞术检测结果，选取20 nmol/L Rapa作为预处理浓度。IDA对THP-1细胞作用24 h的 IC50为59.874 nmol/L。50 nmol/L IDA作用24 h后，Rapa预处理的THP-1细胞增殖抑制率[（69.67±5.03）%比（41.67±3.51）%]和细胞凋亡率[（74.35±4.83）%比（41.25±5.24）%]均高于未预处理的细胞（均 P<0.05）；Rapa预处理的THP-1细胞Beclin-1、LC3 mRNA表达水平及LC3Ⅱ/LC3Ⅰ均高于未预处理的细胞，p62 mRNA表达水平低于未预处理的细胞（均 P<0.05）。 结论:Rapa能增强较低剂量IDA诱导的THP-1细胞凋亡，此效应可能是通过其引起THP-1细胞过度自噬实现的。     

Idarubicin vs doxorubicin in transarterial chemoembolization of intermediate stage hepatocellular carcinoma

BACKGROUND Liver cancer is the fifth most common cancer and the second cause of cancerrelated deaths worldwide.Transarterial chemoembolization(TACE)is the best treatment of intermediate hepatocellular carcinoma(HCC).Doxorubicin is the most commonly used drug despite a low level of evidence.AIM To compare the objective response rate of idarubicin-based TACE(Ida-TACE)against doxorubicin-based TACE(Dox-TACE)in intermediate stage HCC.METHODS Between January 2012 and December 2014,all patients treated with TACE at our academic hospital were screened.Inclusion criteria were patients with Child-Pugh score A or B,a performance status below or equal to 1,and no prior TACE.Either lipiodol TACE or drug-eluting beads TACE could be performed with 10 mg of idarubicin or 50 mg of doxorubicin.Each patient treated with idarubicin was matched with two doxorubicin-treated patients.The TACE response was assessed by independent radiologists according to the mRECIST criteria.RESULTS Sixty patients were treated with doxorubicin and thirty with idarubicin.There were 93%and 87%of cirrhotic patients and 87%and 70%of Child-Pugh A in the doxorubicin and idarubicin groups,respectively.The median number of HCC per patient was two in both groups with 31%and 26%of single nodules in doxorubicin and idarubicin groups,respectively.Objective response rate after first TACE was 76.7%and 73.3%(P=0.797)with 41.7%and 40.0%complete response in doxorubicin and idarubicin groups,respectively.Progression-free survival was 7.7 mo in both groups,and liver transplant-free survival was 24.9 mo and 21.9 mo in doxorubicin and idarubicin groups,respectively.Safety profiles were similar in both groups,with grade 3-4 adverse events in 35%of Dox-TACE and 43%of Ida-TACEs.CONCLUSION Ida-TACE and Dox-TACE showed comparable results in terms of efficacy and safety.Ida-TACE may represent an interesting alternative to Dox-TACE in the management of patients with intermediate stage HCC.     

Treatment with Thalidomide and Cyclophosphamide (TCID) is Superior to Vincristine (VID) and to Vinorelbine (VRID) Regimens in Patients with Refractory or Recurrent Multiple Myeloma

Treatment of relapsed or refractory multiple myeloma remains a challenge and novel treatment regimen are required. Here, a matched pair analysis was performed comparing TCID (thalidomide, cyclophosphamide, idarubicin, dexamethasone) treatment to the treatment of patients with VID (vincristine, idarubicin, dexamethasone) or with VRID (vinorelbine, idarubicin, dexamethasone) for relapsed or refractory multiple myeloma. In total, 197 patients were enrolled in multicenter trials. After matching for important prognostic variables 46 matched-pairs (total of 138 patients) could be analysed with regard to survival, toxicity and efficacy. Interestingly, a significant improvement of overall response rate (ORR) for TCID treatment compared to VID and VRID was found. In addition, TCID treatment also led to a significantly higher overall survival (OS) as well as progression-free survival (PFS) compared to VID and VRID. In conclusion, TCID treatment appears to be superior to VRID and VID treatment in patients with progressive or refractory myeloma.     

Amifostine differentially modulates DNA damage evoked by idarubicin in normal and leukemic cells

Human lymphocytes, p53 protein-deficient acute promyelocytic leukemia cell line HL-60, murine pro-B lymphoid cell line BaF3 and its TEL/ABL-transformed clone cells were exposed to idarubicin with and without pre-treatment with amifostine. Idarubicin at 0.5–5 μM evoked DNA damage measured by the Comet assay. Amifostine at 14 mM decreased DNA-damaging effect of idarubicin in human lymphocytes and BaF3 cells, but increased the effect in TEL/ABL-transformed cells. Amifostine had no influence on the action of idarubicin in HL-60 cells. Our results suggest that the reaction of the cell to DNA damage may contribute to its diverse response to amifostine combined with anticancer drugs and that p53 and fusion tyrosine kinases may be involved in this diversity.     

Increased myelotoxicity of idarubicin: is there a pharmacological basis?

Background: Clinical trials evaluating idarubicin (IDA) in acute myeloid leukemia, multiple myeloma and non-Hodgkin's lymphoma (NHL) have provided some evidence for an increased myelotoxicity of IDA compared to other anthracyclines. IDA is known to be less sensitive towards multidrug resistance mediated by P-glycoprotein (P-gp). This phenotype is a major impediment to successful antineoplastic treatment, but P-gp is also expressed on hematopoietic stem cells (HSC).Methods: We investigated the pharmacokinetics of IDA and etoposide (ETO) in seven previously untreated patients with aggressive NHL. The patients received a CHOP-derived protocol (CIVEP) in which doxorubicin (DOX) was substituted by IDA 11–16 mg/m² and ETO 3×100 mg/m² was added. Furthermore, we evaluated in vitro the impact of P-gp expression on the cytotoxicity of DOX and IDA in cells from three parental chemosensitive leukemia and lymphoma cell lines (HL60, U937, CCRF) and their resistant sublines, as well as in CD34-positive HSC.Results: The peak plasma levels (C_max), terminal elimination half-life (t_1/2) and area under the concentration curve (AUC) both for IDA and for ETO did not differ from published data. In cell line models the numbers of viable cells in a P-gp-expressing resistant CCRF-VCR100 subline were significantly more reduced by IDA (P<0.001), but there was no difference in the cytotoxicities of IDA and DOX in chemosensitive CCRF cells and in the (non-P-gp-expressing) resistant U937 and HL60 sublines. Cytotoxicity against HSC was more pronounced after incubation with IDA than after treatment with DOX (P=0.014), even when a tenfold higher concentration of DOX than of IDA was used. The addition of cyclosporin A increased the cytotoxic effect of DOX but not that of IDA in HSC.Conclusions: The pharmacokinetics of IDA and its main metabolite idarubicinol in CHOP-derived protocols were not different from data obtained with other combinations or monotherapy. The increased myelotoxicity of IDA may be a consequence of P-gp expression in CD34-positive HSC.     

Idarubicin containing regimen in multiple myeloma: preliminary results of a pilot study using a modified "TANDEM" transplant program

Tandem autologous transplant actually represents a challenge in multiple myeloma treatment, but the best conditioning regimen is still under investigation. With the aim of evaluating the feasibility of a modified tandem transplant strategy, we treated 10 multiple myeloma patients after conventional first line chemotherapy with a two step conditioning regimen consisting of high-dose melphalan (200 mg/m 2 ) followed by high-dose melphalan (180 mg/m 2 ) together with indarubicin (15 mg/sqm 2 c.i. ×3 days) both with peripheral stem cell support. At first transplant, the median age was 62 years, performance status was good and disease status was CR in 2 patients and PR in the rest. At the end of the first transplant, 70% of patients achieved CR and only mild toxicity was observed. After the second transplant further improvement of the response rate was obtained with 90% CR. However, we observed three toxic early infection-related deaths from CMV and legionella pneumonia at day +17, +26, +54 after transplantation. Although this schedule seems to be effective in terms of response rate, the 30% TRM imposes an anthracycline dose-reduction with careful patient selection. This approach could reduce the toxic effects and maintain the efficacy of therapy at the same time.     

伊达比星联合阿糖胞苷方案和米托葸醌依托泊苷联合阿糖胞苷方案治疗复发性急性白血病的疗效比较

目的：比较伊达比星(indarubicin)联合阿糖胞苷(cytarabine)(IA方案)和米托蒽醌(mitoxantrone)、依托泊苷(etoposide)联合阿糖胞苷(MEA方案)治疗复发性急性白血病的疗效。方法：将43例复发性急性白血病患者随机分为IA组(21例)和MEA组(22例)，分别应用IA方案或MEA方案进行治疗，比较两组的临床疗效及不良反应。结果：IA组、MEA组的完全缓解率分别为43％、50％，总有效率分别为52％、64％，两组差别无统计学意义。7例复发性急性粒—单核细胞性白血病患者应用MEA方案有4例得到完全缓解。两组均出现严重的骨髓抑制，两组出现的与化疗相关的其它不良反应差别无统计学意义。结论：用IA方案或MEA方案治疗复发性急性白血病均可取得满意的疗效，对急性粒—单核细胞性白血病可首选MEA方案，两种方案均可致严重的骨髓抑制，支持治疗显得很重要。     

去甲氧柔红霉素联合HA方案治疗急性髓细胞白血病初治患者的临床观察

目的:探讨去甲氧柔红霉素(IDA)联合HA方案[(高三尖杉酯碱(HH)加阿糖胞苷(Ara-C)]治疗初发急性髓细胞白血病(AML)的疗效和不良反应。方法:14例AML患者,年龄17～62岁(中位年龄40岁),男8例,女6例,均为初治AML病例。诱导方案为IDA10mg/d,第1～3天,HH3mg/d第1～7天,Ara-C100mg.m-2.d-1第1～7天,静脉滴注。结果:总有效率92.9(13/14),完全缓解率78.6(11/14),治疗过程中未发生早期死亡。化疗的不良反应主要为骨髓抑制和粒细胞缺乏所致感染,未见严重的非造血系统不良反应。结论:IDA联合HA方案为初治AML的高效、安全的方案。