Apoptotic mechanisms in T47D and MCF-7 human breast cancer cells

To investigate the mechanisms underlying apoptosis in breast cancer cells, staurosporine was used as an apoptotic stimulus in the human breast cancer cell lines MCF-7 and T47D. Staurosporine induced dose and time dependent increases in DNA fragmentation which was abrogated by z-VAD-fmk. MCF-7 cells did not express caspase-3, suggesting that DNA fragmentation occurred in the absence of caspase-3 and that other caspases may be involved. Staurosporine induced DEVDase activity in T47D cells suggesting the involvement of caspase-3 and/or caspase-7, yet there was no DEVDase activity in MCF-7 cells, probably ruling out the involvement caspase-7. However, staurosporine induced the cleavage of pro-caspase-6 in MCF-7 cells, but not in T47D cells. Caspase dependent PARP cleavage was detected in MCF-7 cells at 3 h, whereas only partial PARP cleavage was detected in T47D cells and then only after 24 h. Moreover, staurosporine led to cytochrome c release at 2 h in MCF-7 cells and 6 h in T47D cells. In addition, a time dependent and caspase-independent reduction of the mitochondrial transmembrane potential was observed; which appeared to occur after the release of cytochrome c. Translocation of Bax from the cytosol to mitochondria was observed in both cell types, and this preceded cytochrome c release in both T47D and MCF-7 cells. Apoptotic events in both cell types differ temporally, involving activation of different caspases and mitochondrial changes.     

Synergistic interaction between vinorelbine and gamma-linolenic acid in breast cancer cells

It has been suggested that exogenous unsaturated fatty acids (UFAs) may increase the cytotoxic activity of cancer chemotherapeutic agents. We examined how γ-linolenic acid (GLA; 18 : 3n-6), the most promising UFA in the treatment of human tumors, affects the effectiveness of the lipophilic drug vinorelbine (VNR) on human breast carcinoma cell lines. Cells were exposed simultaneously to VNR and GLA or sequentially to GLA followed by VNR. Cell viability was determined by MTT assay. The increase in VNR-induced cell growth inhibition was measured by dividing the IC₅₀ and IC₇₀ values (50 and 70% inhibitory concentrations, respectively) that were obtained when the cells were exposed to VNR alone with those with VNR plus GLA. We found that GLA enhanced in a dose-dependent manner the cell growth inhibitory activity of VNR on MCF-7 cells (up to 9-fold). As GLA by itself showed anti-proliferative effects, possible GLA-VNR interactions at the cellular level were assessed employing the isobologram analysis and the combination index (CI) method of Chou–Talalay. Both methods showed an overall synergism between GLA and VNR in MCF-7 cells. At a high level of cell kill, the synergism was greater when a 24 h GLA pre-exposure or co-exposures were tested. Synergy was likewise observed with the GLA-VNR combination in MDA-MB-231, T47D, and SK-Br3 breast cancer cells. In all cell lines, the synergism was independent of the treatment schedule and the exposure time. Under conditions inhibiting lipid peroxidation using Vitamin E (dl-α-tocopherol), the enhancing effect of GLA (an easily oxidizable UFA) on VNR activity was partially abolished. However, when Vitamin E was used in combination, a similar synergistic increase in growth inhibition was obtained. These latter observations strongly implies that the synergistic effects of GLA with VNR are not mediated through a mechanism involving a generation of lipoperoxides. For comparison, the effects of other UFAs were examined on VNR chemosensitivity: GLA was the most potent at enhancing VNR activity, followed by docosahexaenoic acid (22 : 6n-3), eicosapentaenoic acid (20 : 5n-3) and α-linolenic acid (18 : 3n-3), whereas linoleic acid (18 : 2n-6) and arachidonic acid (20 : 4n-6) did not increase VNR chemosensitivity. Very high concentrations of oleic acid (OA; 18 : 1n-9), an UFA inversely correlated with breast cancer risk, also enhanced VNR effectiveness. Thus, various types of UFAs were not equivalent with respect to their actions on VNR effectiveness. In conclusion, our results give experimental support to the hypothesis that some UFAs can be used as modulators of tumor cell chemosensitivity and provide the rationale for in vivo preclinical investigation.     

Apoptosis signalling mechanisms in human cancer cells induced by Calphostin-PDT

Photodynamic therapy (PDT) is a promising treatment that is approved by the US FDA for the treatment of oesophageal and lung cancer as well as for age-related macular degeneration. In this study, using standard tissue culture techniques, the photo cytotoxicity and apoptotic mechanisms of Calphostin C (Cal C), a perylenequinone microbial compound in combination with visible light dose was examined in different tumor cell lines. Our results demonstrated both a time and drug-light dose dependence in Cal-C-PDT induced photo toxicity and apoptotic cell death. The induction of apoptosis by Cal C-PDT was found to transit to necrotic cell death at higher drug and light doses. The detection of apoptosis in irradiated tumor cells was performed using various approaches including cell morphology analysis, flow cytometry [DNA fragmentation and phosphatidylserine (PS) externalization] and biochemical assays (activation of caspases). Time-course analysis of Cal C cellular uptake and distribution showed a rapid increase within the cellular compartments. The activation of caspases and nuclear fragmentation was evidenced at a maximum time point of 3 h after irradiation. By the use of specific caspase substrates, significant activation of caspase-8 and -3 was found. Mitochondrial involvement during Cal C-PDT-induced apoptosis was proven by a rapid reduction of the mitochondrial membrane potential. Furthermore, Cal C-PDT also enhanced FasL expression, which then induced Fas signalling-dependent cell death in NPC and colon cancer cell lines tested. Our results contribute to a deeper understanding of the processes involved in apoptotic cell death following photodynamic treatment with Cal C.     

Capecitabine and vinorelbine in metastatic breast cancer

BackgroundAs anthracyclines and taxanes are frequently used in the adjuvant and first-line metastatic settings, capecitabine and vinorelbine are frequently used as monotherapy and in combination for metastatic breast cancer (MBC). In the absence of comparative, phase III data, retrospective analyses and cross-trial comparisons provide the only indication of the relative efficacy of these options.MethodsWe reviewed studies evaluating the 2 agents alone or in combination in MBC.ResultsWe identified 6 capecitabine and 2 vinorelbine phase III trials, numerous phase II monotherapy studies and 35 phase I/II studies exploring capecitabine–vinorelbine combination therapy (1 with trastuzumab in HER2-positive MBC).ConclusionFor monotherapy, the limited, retrospective comparative evidence supported by consistent prospective data suggests that capecitabine is more effective than vinorelbine. Comorbidities, organ function tolerability, tumour biology and patient characteristics should also inform treatment choice. If combination therapy is deemed clinically appropriate, intravenous vinorelbine with capecitabine may be considered, potentially improving efficacy compared with monotherapy, but at the cost of increased toxicity. Randomised evaluation versus capecitabine monotherapy is ongoing. In contrast, cross-trial comparison suggests that addition of oral vinorelbine to capecitabine adds haematological toxicity without apparently improving efficacy in pretreated MBC. Data from small, single-arm, phase II studies in the first-line setting are more encouraging. In summary, the strongest clinical data support capecitabine monotherapy in the majority of patients. In certain populations, a capecitabine–vinorelbine combination may be appropriate but requires further validation in randomised trials.     

In vitro effects of trastuzumab and vinorelbine in trastuzumab-resistant breast cancer cells

Purpose The majority of patients who initially respond to trastuzumab will progress within 1 year. Currently, patients who progress after trastuzumab-based therapy are often maintained on trastuzumab combined with a different chemotherapeutic agent, such as vinorelbine. However, evidence supporting the continued use of trastuzumab in these breast cancers is lacking.Methods We created a preclinical model of trastuzumab resistance using the SKBR3 HER-2-overexpressing breast cancer cell line. Dose-response and cell cycle alterations in response to trastuzumab and/or vinorelbine were assessed.Results In contrast to the parental SKBR3 cells, vinorelbine-mediated growth inhibition and apoptosis were not significantly enhanced by the addition of trastuzumab in the trastuzumab-resistant pools.Conclusions These results suggest that the continued treatment of trastuzumab-resistant breast cancers with trastuzumab-containing regimens may not be effective. A randomized clinical trial of trastuzumab plus vinorelbine versus vinorelbine alone should be conducted in patients with HER-2-overexpressing breast cancer to determine the optimal duration of trastuzumab therapy upon progression.     

Mechanism of the radiosensitization induced by vinorelbine in human non-small cell lung cancer cells.

Vinorelbine (Navelbine, KW-2307), a semisynthetic vinca alkaloid, is a potent inhibitor of mitotic microtubule polymerization. The aims of this study were to demonstrate radiosensitization produced by vinorelbine in human non-small cell lung cancer (NSCLC) PC-9 cells and to elucidate the cellular mechanism of radiosensitization. A clonogenic assay demonstrated that PC-9 cells were sensitized to radiation by vinorelbine with a maximal sensitizer enhancement ratio at a 10% cell survival level of 1.35 after 24-h exposure to vinorelbine at 20 nM. After 24-h exposure to vinorelbine at 20 nM, the approximately 67% of the cells that had accumulated in the G2/M-phase were cultured in the absence of vinorelbine and then irradiated at a dose of 8 Gy. Flow cytometric analyses showed prolonged G2/M accumulation concomitant with continuous polyploidization, and induction of apoptosis was observed in the cells subjected to the combination of vinorelbine-pretreatment and radiation. Polyploidization and induction of apoptosis were confirmed by morphological examination and a DNA fragmentation assay, respectively. We concluded that vinorelbine at a minimally toxic concentration moderately sensitizes human NSCLC cells to radiation by causing accumulation of cells in the G2/M-phase of the cell cycle. Prolonged G2/M accumulation concomitant with continuous polyploidization and increased susceptibility to induction of apoptosis may be associated with the cellular mechanism of radiosensitization produced by vinorelbine.     

姜黄素诱导人胃癌细胞SGC-7901凋亡的作用机制

 目的 探讨姜黄素诱导人胃癌细胞SGC-7901凋亡的作用及其相关机制。方法 以不同浓度的姜黄素作用于胃癌SGC-7901细胞，利用倒置相差显微镜观察细胞生长形态变化，通过MTT检测细胞生长抑制率，流式细胞术检测细胞凋亡，Western blot 检测胃癌细胞中Fas及survivin的表达情况。结果 姜黄素能显著抑制体外培养的SGC-7901细胞的生长并呈量-效和时-效关系，流式细胞术检测到亚二倍体凋亡峰，细胞凋亡率增加，Western blot结果提示经姜黄素作用后Fas表达率上升，survivin表达率下降。结论 姜黄素能抑制胃癌细胞SGC-7901的生长并促进其凋亡，姜黄素可能通过上调Fas及下调survivin的表达而诱导凋亡。     

单药长春瑞滨对乳腺癌原代细胞杀伤效果的体外研究

 目的 探讨长春瑞滨用于复发性乳腺癌患者的治疗效果和应用前景。方法 取得原发性和复发性乳腺癌患者的癌组织标本，采用胶原酶消化法获取乳腺癌原代细胞，应用原代培养和MTT法检测5种化疗药物在体外对乳腺癌原代细胞的杀伤效果。结果 对原发性乳腺癌原代细胞，紫杉醇和多西紫杉醇（多西他赛）的杀伤效果（敏感率分别为91.04 ％、92.54 ％）优于多柔比星、表柔比星和长春瑞滨（敏感率分别为73.13 ％、74.63 ％、71.64 ％；P＜0.01）；对复发性乳腺癌原代细胞，长春瑞滨和紫杉醇、多西紫杉醇的杀伤效果（敏感率分别为74.07 ％、66.67 ％、66.67 ％）明显优于多柔比星和表柔比星（敏感率分别为37.04 ％、40.74 ％；P＜0.01）。结论 长春瑞滨可以作为化疗一线药物应用于复发性乳腺癌患者。     

Prevention of phlebitis caused by vinorelbine chemotherapy in outpatients with breast cancer

We studied the prevention of phlebitis in 10 patients who had developed the symptoms after receiving vinorelbine to treat breast cancer at our outpatient chemotherapy clinic from July 2005 to August 2006. Veins proximal to the injection site were warmed using hot compresses during the vinorelbine injection and physiological saline was increased to wash out the drug after the injection from 250 mL to 500 mL in combination to investigate whether the treatment was effective in preventing phlebitis. The severity of phlebitis was significantly decreased after the combined treatment compared with the pre-treatment level(p=0.039). The combination was effective to relieve vascular pain during the injection in all 10 patients, and the number of event occurrences was significantly decreased(p<0.0005). It was also effective to decrease the frequency of vascular pain after patients returned home(p=0.001). The combination of hot compresses and increase of physiological saline for washing out was an effective treatment to prevent phlebitis caused by vinorelbine. The comparison of patient characteristics to find other contributing factors to phlebitis than vinorelbine revealed no association with the number of doses, diameter of the vein to be punctured, or pretreatment.     

Weekly schedule of vinorelbine in pretreated breast cancer patients

Purpose: In this phase II study, we explored tolerability and activity of vinorelbine administered according to a dose-dense weekly schedule with hematopoietic growth factor support in pretreated, advanced breast cancer patients. Patients and Methods: From January 1994 to March 1996, 40 patients with metastatic breast cancer, pretreated with at least one prior anthracycline-containing regimen, were entered into the study. Patient characteristics: median age 53 years (range 32–70); ECOG performance status 0-1: 34 patients, 2: 6 patients; dominant visceral metastatic disease: 15 patients, dominant non-visceral: 25; anthracycline-refractory/resistant: 2 patients, sensitive: 38 patients. Six patients were treated as first-line therapy for metastatic disease and 34 in second- or subsequent lines.All patients received vinorelbine at the dose of 25 mg/m²/week as a short intravenous infusion, together with routine antiemetic medication. Granulocyte-colony stimulating factor (Lenograstim) at the dose of 150 g/m² subcutaneously on day 3 was included in the treatment schedule. Results: The median number of treatment weeks was 23 (range: 4–24), with a delivered dose-intensity (DDI) of 23.8 mg/m²/week (range: 18.7–25, 95.2% of projected dose-intensity).Toxicity was mild, with non-complicated neutropenia being the main toxicity observed (grade 3–4 in 25% of the patients but only 2% of treatment weeks). Overall response rate was 52.5%, with complete responses in 12.5% of patients. Median duration of the response and median time to progression were 10 and 9 months, respectively. Median overall survival was 19 months. Conclusion: Dose-dense weekly vinorelbine is safe and effective with minimal toxicity in pretreated advanced breast cancer patients.     

Efficacy of vinorelbine in advanced and metastatic breast cancer

Vinorelbine is a new anti-cancer drug that is available for advanced or metastatic breast cancer, approved by the Japanese Ministry of Health, Labour and Welfare in May 2005. We evaluated its efficacy and safety in 35 patients treated with vinorelbine since April of 2005 to February of 2009. Patient's average age was 52 years old, and the average number of previous treatments was 2. 7. The response rate was 8. 6%; there was no complete responder, and three partial responders. Median duration of response was 5. 3-months. Clinical benefit rate was 28. 6%, 16. 7% in the vinorelbine monotherapy group, and 54. 3% in the VNR/trastuzumab combination therapy group. The adverse event was observed in 5. 7% as grade 3 or 4 neutropenia, and in 2. 9% as Grade 1 superficial phlebitis. These results suggest that vinorelbine is a safe and effective agent among a limited number of patients.     

Combination of trastuzumab and vinorelbine in metastatic breast cancer

BACKGROUND: Since the clinical introduction of trastuzumab (Herceptin) for metastatic breast cancers that overexpress human epidermal growth factor receptor 2 (HER2), this anticancer agent has played an important role in breast cancer treatment. We examined the effects of trastuzumab and vinorelbine (Navelbine) as a second- or third-line therapy in 24 patients whose HER2-positive tumors did not respond to or relapsed after administration of trastuzumab alone or in combination with taxane. METHODS: Trastuzumab was administered at 2 mg/kg (loading dose 4 mg/kg) once weekly and vinorelbine at 25 mg/m(2) once weekly. The median treatment duration was 118.5 days (range, 22-351 days). RESULTS: The response rate was 42% (95% confidence interval (CI): 22%-63%). The adverse events of NCI-CTC grade 3 or above consisted of neutropenia in three patients; other adverse events, including vasculitis, generalized fatigue, anemia and thrombocytopenia, were grade 1 or 2. All adverse events were reversible after treatment withdrawal and were easily manageable. CONCLUSION: A combination of trastuzumab and vinorelbine can be safely administered on an outpatient basis, and is useful in the treatment of patients with HER2-overexpressing metastatic breast cancer.     

Combination of vinorelbine + doxorubicin in advanced breast cancer

OBJECTIVE: To assess the efficacy of a vinorelbine + doxorubicin combination in terms of response rate and time to progression in patients with locally advanced or metastatic breast cancer. METHODS: Vinorelbine (25 mg/m2) and doxorubicin (25 mg/m2) were administered intravenously in a rapid injection on days 1 and 8 every 21 days. Initially, 3 courses of vinorelbine + doxorubicin were given. Patients with responding or stable disease received 6 more courses to a maximum of 9 courses. RESULT: Twenty-nine patients were entered into the study and 27 eligible patients were considered evaluable for response. Median age was 45 years (range 33 to 63). Overall response rate was 66.67% (18/27) (CR = 5, PR = 13). Median time to progression was 7.8 months (range 4 to 16) and the median survival time was 25.9 months. Median follow-up time was 8.5 months (range 1.5 to 25). Toxicity was generally moderate. Hematologic complication was the dose limiting toxicity. WHO grade III/IV neutropenia was observed in 18.5%/3.7% of patients. The major non-hematologic toxicities were nausea and phlebitis. Grade III nausea/vomiting was observed in 7.4% and grade III/IV phlebitis in 3.7%/3.7% of patients. No toxic deaths were observed. CONCLUSION: The present vinorelbine + doxorubicin combination was highly effective and generally well tolerated in cases of advanced breast cancer. Further studies are required.     

瘦素诱导乳腺癌细胞的凋亡抵抗及其分子机制

Objective To explore the apoptosis resistance induced by Leptin and its mechanism in breast cancer cells in vitro.Methods The leptin-mediated reduction of docetaxel-induced apoptosis in human breast cancer T47D cells was evaluated by TransAM ELISA,MTT and caspase-9 assay.The leptinpromoted survivin expression was analyzed by Western-blot and RT-PCR.The reversing effect of STAT3 knockdown on leptin-induced survivin upregulation was measured by Western-blot and RT-PCR.Results Leptin promoted T47D cells proliferation and the inhibitory rate was-63.6%.It reduced docetaxel-induced apoptosis in T47D cells by 31.9%.Leptin at different concentrations promoted survivin protein and mRNA expression in T47D cells.The expression of survivin mRNA was 4.6 fold compared with the T47D cells not treated with leptin(10 nmol/L).The expression of survivin mRNA in T47D cells was 0.55±0.15 fold after transfected with small interfering RNA(siRNA)of STAT3.The expression of survivin mRNA in STAT3 siRNA group and mock transfected group were 0.56±0.18 fold and 1.61±0.22 fold after treated by leptin,respectively.The survivin protein level of T47D mock transfected cells was increased after treated by leptin,but the protein level of T47D transfected with STAT3 siRNA cells were not changed significantly.Conclusion Leptin/STAT3 signaling is a novel pathway for up-regulation of survivin expression in breast cancer cells.     

瘦素诱导乳腺癌细胞的凋亡抵抗及其分子机制

目的 观察瘦素对乳腺癌细胞凋亡的影响,并探讨其分子机制.方法 采用TransAM酶联免疫吸附法(ELISA)检测瘦素对乳腺癌细胞T47D凋亡的影响,采用四甲基偶氮唑蓝(MTT)法检测细胞毒性,并测定T47D细胞caspase-9的活性.采用实时荧光定量逆转录聚合酶链反应(RT-PCR)和Western blot法测定瘦素对T47D细胞survivin mBNA和蛋白表达水平的影响.采用BNA干扰技术沉默信号转导和转录激活因子3(STAT3)基因,并采用实时荧光定量逆转录聚合酶链反应(RT-PCR)和Westem blot法测定瘦素对沉默STAT3基因后T47D细胞survivin mRNA和蛋白表达水平的影响.结果 瘦素可以促进人乳腺癌细胞T47D的增殖,增殖抑制率为63.6%;减少多西他赛诱导的细胞凋亡,达31.9%.不同浓度瘦素均可促进T47D细胞survivin mRNA的表达,其中以10 nmol/L时作用最明显.10 nmol/L瘦素作用60 min后,T47D细胞中survivin mRNA的表达量是瘦素作用前的4.6倍;作用2 h后,T47D细胞中survivin蛋白的表达增强.将STAT3小干扰RNA(siRNA)转入T47D细胞30 min后,survivin mRNA表达的变化倍率为0.55±0.15.经瘦素处理后,STAT3 siRNA质粒转染组和空质粒对照组survivin mRNA表达的变化倍率分别为0.56±0.18和1.61±0.22.将STAT3 siRNA转入T47D细胞后,survivin蛋白表达下降.经瘦素处理后,转染空质粒的细胞survivin蛋白的表达增加,而转入STAT3 siRNA的细胞survivin蛋白无明显变化.结论 在乳腺癌T47D细胞中,瘦素/STAT3信号通路是上调survivin表达的有效途径.     

A review of vinorelbine in the treatment of breast cancer

As combinations and sequences of anthracyclines and taxanes increasingly become standard adjuvant treatment for early breast cancer, a major need for new treatment options for metastatic breast cancer will arise. Vinorelbine is highly active in the treatment of metastatic breast cancer, both as a single agent and in combination regimens. Furthermore, it is well tolerated, with a low incidence of subjective toxicities. It is anticipated, therefore, that vinorelbine will become increasingly utilized for treating metastatic breast cancer due to its favorable safety profile, good tolerability, and promising results in combination with other chemotherapy agents. Combinations with trastuzumab and newer molecular targeting agents are being explored. Doublets or triplets of vinorelbine with drugs other than anthracyclines and taxanes could be considered in the next generation of adjuvant and neoadjuvant trials, where it is anticipated that anthracycline/taxane combinations are likely to replace anthracycline/cyclophosphamide combinations as the mainstay of adjuvant treatment.     

洛铂联合长春瑞滨治疗晚期乳腺癌33例的临床疗效

目的:观察长春瑞滨(NVB)联合洛铂(LBP)组成NL方案治疗晚期乳腺癌的疗效和不良反应。方法:从2004年1月~2005年10月,采用NVB加LBP的联合化疗方案治疗晚期乳腺癌33例,LBP30mg/m2,第1天,静脉滴注3小时;NVB25mg/m2,第1、8天,静脉滴注30分钟,21天为1周期。结果:全组33例共完成105个周期,中位3周期(2~4周期),均可评价疗效。获CR1例,PR13例,SD11例,PD8例,RR为42·4%,肿瘤控制率DCR(CR PR SD)为75·8%,中位缓解期为4个月(1~17个月)。主要不良反应为骨髓抑制,其中Ⅲ~Ⅳ度粒细胞减少发生率为57·6%,Ⅲ度血小板减少发生率为9·1%;非血液学毒性轻微,可以耐受。结论:洛铂联合长春瑞滨组成NL方案治疗局部晚期及转移性乳腺癌疗效较好,不良反应可以耐受,在加强支持治疗(应用造血因子)的基础上,可以考虑作为二线治疗或解救化疗方案。     

Mitoxantrone plus vinorelbine in pretreated patients with metastatic breast cancer

Vinorelbine and mitoxantrone have both been demonstrated to have significant antitumor activity in patients with breast cancer. The aim of this study was to evaluate the efficacy and safety of the combination as second or third line treatment in patients with metastatic breast cancer (MBC). Fifty-one previously treated patients with MBC were enrolled from October 2001 to May 2004 and 48 were eligible for evaluation. Median age was 59 years (range 33-82) and ECOG performance status was < or =2. Distant sites of metastasis were as follows: liver 64%, bone 49%, lung 36%, lymph nodes 6%, skin 4%, brain 2% and other sites 6%. All patients received vinorelbine 20 mg/m(2), D1+8 and mitoxantrone 10 mg/m(2) D8 every 21 days for 6 cycles. All eligible patients were analyzed for toxicity and response. Two patients (4%) achieved complete response and 12 (25.5%) partial response. The objective overall response rate was 29.5% (95% confidence interval [CI] 17 - 45), 9 (19%) patients had stable disease, 17 (36%) had progressive disease and 7 (15%) were non-evaluable. After a median follow up of 18 months, overall survival was 13 months (range 0.8 - 38+) and median time to disease progression was 5 months (range 1 - 32). A total of 280 cycles was delivered. The relative dose intensities of mitoxantrone and vinorelbine were 79% and 77%, respectively. Toxicities (grade III-IV) were as follows: leukopenia 18 (38%), neutropenia 21 (45%), thrombocytopenia 1 (2%), anemia 4 (8.5%), alopecia 2 (4%) and constipation 1 (2%). Febrile neutropenia was recorded in one patient. There were no treatment related deaths. The combination of mitoxantrone and vinorelbine is an effective regimen with manageable toxicity in pretreated patients with advanced breast cancer.