Paclitaxel and cyclosporine A show supra-additive antiproliferative effects on smooth muscle cells by activation of protein kinase C

We intended to establish a pharmacologic concept of synergistic antiproliferative effects on smooth muscle cells (SMC) by using paclitaxel and cyclosporine A at clinically applicable doses. Coronary SMC were incubated with paclitaxel and cyclosporine A at concentrations of 10 – 20 nmol/L and 83 – 415 nmol/L, respectively. Antiproliferative effects were assessed by cell counts, [³H]thymidine incorporation and cell cycle analysis. In addition, apoptosis was studied by cytoplasmic histone-associated DNA fragments and in vitro protein kinase C activity (PKC) was determined by immunoassay. We found paclitaxel and cyclosporine A to exert a highly supra-additive antiproliferative effect on SMC with significant reductions of cell counts (p < 0.01) and [³H]thymidine incorporation (p < 0.05). SMC were found to be arrested at the G2/M transition. This antiproliferative effect was observed in the absence of DNA fragmentation above values obtained for single compound treatment, which had virtually no impact on cell proliferation. DNA fragmentation started to increase at a drug combination comprising paclitaxel at the higher dose of 20 nmol/L. Under the treatment with both paclitaxel and cyclosporine A, PKC activity showed a 1.8-fold increase (p < 0.05) compared with untreated controls. In conclusion, PKC mediates supra-additive antiproliferative effects of paclitaxel and cyclosporine A on SMC. The data demonstrate a highly efficient pharmacologic concept for the inhibition of SMC proliferation. Further studies are needed to test this concept under in vivo conditions for the prevention of restenosis or transplant vasculopathy by systemic application of cyclosporine A – when already applied for immunosuppressive purposes – and local delivery of paclitaxel. Received: 28 August 2001, Returned for revision: 25 September 2001, Revision received: 20 November 2001, Accepted: 4 December 2001     

A phase I study of paclitaxel, topotecan, cisplatin and Filgrastim in patients with newly diagnosed advanced ovarian epithelial malignancies: a Gynecologic Oncology Group study

OBJECTIVES: To determine a recommended dose level (RDL) of paclitaxel, cisplatin and topotecan in women with previously untreated epithelial ovarian or peritoneal cancer as a possible experimental arm in a future Gynecologic Oncology Group phase III study. METHODS: Patients with newly diagnosed stage III or IV disease were treated with paclitaxel 175 mg/m2/3 h, followed 2 h later by cisplatin 50 mg/m2 on day 1. Topotecan was administered on consecutive days as a 30-minute infusion, beginning after cisplatin on day 1, receiving either 5 days beginning at 0.3 mg/m2 (cohort 1), or 3 days beginning at 0.5 mg/m2 (cohort 2). Treatment was given every 21 days for a maximum of 8 cycles. RESULTS: Forty-five evaluable patients were enrolled in the two cohorts. Thrombocytopenia and prolonged neutropenia were the major dose-limiting toxicities. Dose-limiting neutropenia was seen at the first dose level, thus all subsequent dose escalations included Filgrastim. The RDL of cohort 1 was paclitaxel 175 mg/m2/3 h, cisplatin 50 mg/m2 and topotecan 0.5 mg/m2 daily x 5 with Filgrastim. The RDL of cohort 2 was paclitaxel 175 mg/m2/3 h, cisplatin 50 mg/m2 and topotecan 0.75 mg/m2 daily x 3 with Filgrastim. CONCLUSION: In women with previously untreated epithelial ovarian or peritoneal cancer the combination of paclitaxel, cisplatin and topotecan is feasible. However, this treatment requires the use of Filgrastim and attenuated dosing of topotecan in both a 5-day and 3-day topotecan infusion schedule.     

生脉注射液联合化疗治疗晚期非小细胞肺癌的临床研究

目的:观察生脉注射液联合泰素加卡铂方案治疗晚期非小细胞肺癌的临床效果及其减毒作用。方法:将38例晚期非小细胞肺癌患者随机分为治疗组和对照组各19例。对照组第1天予泰素135 mg/m2,3 h静滴,卡铂AUC 5静滴,21 d为1个周期;治疗组予同一化疗方案并于化疗结束后第2天用生脉注射液60 m l加入50 g/L葡萄糖注射液500 m l静滴,每天1次,连用15 d。两个周期后按照WHO实体瘤近期客观疗效评定标准进行评价。结果:治疗组和对照组近期有效率分别为52.6%和47.4%;治疗组Karnorfsky评分增加6例,对照组增加3例;治疗组体质量明显增加,骨髓抑制发生率明显较对照组低。结论:生脉注射液能提高泰素加卡铂方案化疗患者的生存质量,减轻化疗后骨髓抑制。     

胃癌腹膜转移病人腹腔化疗港并发症发生及危险因素分析

目的:分析腹腔化疗港并发症发生及相关危险因素.方法:回顾性分析我院323例放置腹腔化疗港病人的临床资料,纳人261例胃癌腹膜转移病人.分析腹腔化疗港在胃癌腹膜转移病人腹腔化疗时发生的并发症及其危险因素.结果:261例中59例(22.6％)发生化疗港相关并发症.其中,皮下积液(25例,42.4％)和化疗港感染(16例,2...     

紫杉醇联合顺铂治疗晚期乳腺癌30例临床观察

目的评价紫杉醇联合顺铂治疗晚期乳腺癌的疗效和毒副作用。方法符合人组条件的晚期乳腺癌31例,化疗前常规抗过敏治疗,化疗方案紫杉醇（TAX）175mg／m^2静脉滴注d1,顺铂（DDP）30mg／m^2静脉滴注d1-3,21d为1周期,化疗3周期。结果30例完成疗效观察,RR56．7％（17例）,CR2例（6．7％）,PR15例（50．0％）。主要毒副作用为骨髓抑制,胃肠道反应,肌肉关节痛和周围神经痛。白细胞减少的发生率96．7％,其中Ⅲ度10．0％,无Ⅳ度发生。恶心、呕吐的发生率90．0％,无Ⅲ-Ⅳ度发生。结论紫杉醇联合顺铂是一个疗效肯定、毒副作用可耐受的方案。     

Inhibitory effects of everolimus in combination with paclitaxel on adriamycin-resistant breast cancer cell line MDA-MB-231

ObjectiveWe aimed to evaluate the therapeutic effects of paclitaxel in combination with mTOR inhibitor everolimus on adriamycin-resistant breast cancer cell line MDA-MB-231 (MDA-MB-231/ADR).Materials and methodsMDA-MB-231/ADR cells were treated with different concentrations of paclitaxel and everolimus. The IC₅₀ values after 48 h of treatment were measured by the MTT assay. The apoptosis rate and cell cycle were detected by flow cytometry. The protein expressions of Akt, PI3K, mTOR, p-pI3K, p-AKT and p-mTOR were detected by Western blot.ResultsWhen paclitaxel at ≥1.56 μg/ml was used, the growth of MDA-MB-231/ADR cells was inhibited more significantly than that of control group (P < 0.05). After treatment with ≥6.25 μg/ml everolimus, the cell growth was also suppressed more significantly (P < 0.05). The IC₅₀ values of everolimus and paclitaxel were 32.50 μg/ml and 7.80 μg/ml, respectively. The inhibition rate of paclitaxel plus everolimus was significantly enhanced with increasing paclitaxel concentration (P < 0.001). After treatment with 7.80 μg/ml paclitaxel, the two drugs had best synergistic inhibitory effects on proliferation. Compared with drugs alone, the combination significantly promoted apoptosis (P < 0.001). The paclitaxel + everolimus group had significantly more cells in the G0-G1 phase than those of control and individual drug groups (P < 0.001). Everolimus significantly decreased mTOR and p-mTOR expressions compared with those of control group (P < 0.001). Compared with everolimus alone, the combination reduced the expressions more significantly (P < 0.05). Paclitaxel decreased the expression levels of PI3K, p-PI3K and p-AKT. Compared with paclitaxel alone, the combination significantly promoted the reduction of PI3K, p-PI3K and p-AKT expressions (P < 0.05).ConclusionEverolimus can enhance the effect of paclitaxel on MDA-MB-231/ADR cells, inhibit cell proliferation, induce apoptosis and arrest cell cycle in the G1 phase mainly by down-regulating the expressions of key proteins in the mTOR signaling pathway.     

Randomized Phase II Study of Paclitaxel plus Alisertib versus Paclitaxel plus Placebo as Second-Line Therapy for SCLC: Primary and Correlative Biomarker Analyses

IntroductionWe assessed the Aurora A kinase inhibitor, alisertib, plus paclitaxel (henceforth referred to as alisertib/paclitaxel) as second-line treatment for SCLC.MethodsIn this double-blind study, patients with relapsed or refractory SCLC were stratified by relapse type (sensitive versus resistant or refractory) and brain metastases and randomized 1:1 to alisertib/paclitaxel or placebo plus paclitaxel (henceforth referred to as placebo/paclitaxel) in 28-day cycles. The primary end point was progression-free survival (PFS). Associations of c-Myc expression in tumor tissue (prespecified) and genetic alterations in circulating tumor DNA (retrospective) with clinical outcome were evaluated.ResultsA total of 178 patients were enrolled (89 in each arm). The median PFS was 3.32 months with alisertib/paclitaxel versus 2.17 months with placebo/paclitaxel (hazard ratio [HR] = 0.77, 95% confidence limit [CI]: 0.557–1.067, p = 0.113 in the intent-to-treat population versus HR = 0.71, 95% CI: 0.509–0.985, p = 0.038 with corrected analysis applied). Among 140 patients with genetic alternations, patients with cell cycle regulator mutations (cyclin-dependent kinase 6 gene [CDK6], retinoblastoma-like 1 gene [RBL1], retinoblastoma-like 2 gene [RBL2], and retinoblastoma 1 gene [RB1]) had significantly improved PFS with alisertib/paclitaxel versus with placebo/paclitaxel (3.68 versus 1.80 months, respectively [HR = 0.395, 95% CI: 0.239–0.654, p = 0.0003]), and overall survival (7.20 versus 4.47 months, respectively [HR = 0.427, 95% CI: 0.259–0.704, p = 0.00085]). A subset of patients with c-Myc expression showed significantly improved PFS with alisertib/paclitaxel. The incidence of grade 3 or higher drug-related adverse events was 67% (58 patients) with alisertib/paclitaxel versus 22% (25 patients) with placebo/paclitaxel. Twelve patients (14%) versus 11 (12%) died on study, including four versus zero treatment-related deaths.ConclusionsEfficacy signals were seen with alisertib/paclitaxel in relapsed or refractory SCLC. c-Myc expression and mutations in cell cycle regulators may be potential predictive biomarkers of alisertib efficacy; further prospective validations are warranted.     

TA方案和NP方案治疗晚期乳腺癌临床研究

目的：观察比较紫衫醇＋表阿霉素（TA）和长春瑞滨＋顺铂（NP）两种化疗方案治疗晚期乳腺癌的疗效和毒性.方法：46例晚期乳腺癌患者随机分为TA组和NP组,每组23例.TA组紫杉醇135 mg/m^2,第1天;表阿霉素25 mg/m^2 ,第2～3天.NP组长春瑞滨40 mg/m^2,第1天、第8天;顺铂25 mg/m^2, 第1～4天. 结果：疗效按WHO标准,TA组总有效率65.2%,NP组总有效率为56.5%,两组间比较差异无统计学意义（P＞0.05）.主要毒副反应中,骨髓抑制、恶心呕吐、脱发,两组间差异无统计学意义（P＞0.05）;TA组的关节肌肉酸痛发生率（39.1%）高于NP组（4.3%）,NP组的静脉炎发生率（43.5%）,明显高于TA组（8.7%）,组间比较均有统计学意义 （P＜0.05）.结论：TA和NP两组方案治疗复发转移性乳腺癌疗效高且无统计学差别,均可作为治疗晚期乳腺癌的有效方案.     

hTERT反义寡核苷酸促进紫杉醇诱导Hut78细胞凋亡

目的：探讨转染人端粒酶逆转录酶基因（hTERT)反义寡核苷酸后能否增强紫杉醇诱导皮肤T细胞淋巴瘤细胞株Hut78凋亡。方法:采用脂质体介导的基因转染技术将hTERT反义寡核苷酸（AODN)、正义寡核苷酸（SODN)导入Hut78细胞株中，应用端粒酶重复序列扩增及酶联免疫吸附方法（TRAP-PCR-ELISA)、逆转录-聚合酶链反应、四甲基偶氮唑蓝法（MTT)、台盼蓝拒染法和流式细胞仪分析测定联合应用紫杉醇后对细胞端粒酶活性、hTERT mRNA表达、细胞增殖及凋亡的影响。结果:转染hTERT基因反义寡核苷酸并联合应用紫杉醇，48 h后Hut78细胞增殖即被明显抑制，端粒酶活性下降，hTERT mRNA表达降低，分别同SODN联合紫杉醇组及单用紫杉醇组进行比较，有显著差异（P<0.05)； MTT结果显示，紫杉醇对AODN组、SODN组及空白对照组细胞的IC50值分别为(81.10 ± 1.68) nmol/L，(138.70±1.80) nmol/L和(139.40± 5.50) nmol/L，差异显著（P< 0.05)；经流式细胞仪检测，AODN联合紫杉醇组凋亡率明显高于SODN联合紫杉醇组和单用紫杉醇组。结论:hTERT反义寡核苷酸与紫杉醇有协同抗肿瘤效应，并能增加Hut78细胞对紫杉醇的敏感性，促进紫杉醇诱导Hut78细胞凋亡。     

超声破坏载紫杉醇微泡对卵巢癌细胞株SKOV3的抑制增殖及诱导凋亡作用

目的探讨超声破坏载紫杉醇微泡后对卵巢癌细胞株SKOV3生长抑制及凋亡诱导作用。 方法体外培养卵巢癌细胞,将细胞分为5组,即紫杉醇组,紫杉醇联合超声辐照组,载紫杉醇微泡联合超声辐照组,单纯载紫杉醇微泡组及空白对照组。用MTT法观察超声破坏载紫杉醇微泡后在不同时间对细胞的生长抑制情况,用透射电镜观察细胞的凋亡诱导情况。 结果在紫杉醇浓度为1×10^-6mol/L时,载紫杉醇微泡联合超声组的细胞在微泡辐照击破后生长明显受到抑制,并且随培养时间延长,抑制效应越明显,透射电镜观察到载紫杉醇微泡联合超声组的细胞有凋亡小体,比其它各组对卵巢癌细胞株SKOV3的凋亡诱导作用强。 结论载紫杉醇微泡被超声辐照破坏后对卵巢癌细胞株SHKOV3生长有明显抑制作用,并且能诱导其凋亡。