人多发性骨髓瘤耐药细胞系MOLP-2/R的建立及其耐药机制

目的 建立耐马法兰的人多发性骨髓瘤细胞系MOLP-2/R,并对其生物学特性及耐药机制进行初步探讨.方法 采用马法兰浓度梯度递增法,建立耐马法兰多发性骨髓瘤细胞系.检测两种细胞的形态差异、生长曲线及倍增时间;用药物敏感试验检测两种细胞对马法兰及多种化疗药物的半数抑制浓度(JC50)和耐药指数(RJ);使用Western blot比较两种细胞P_gP、MRP和FANCD2的表达差异来探讨可能的耐药机制.结果 成功建立了耐药指数为6.03的M()Lp2/R耐药细胞系,与MOLP-2细胞相比,MOLP-2/R耐药细胞异形明显;耐药细胞倍增时间显著延长(P<0.05);MOLP_2/R且对ADM、CTX、DDP、VP-16有交叉耐药;MOLp-2/R中FANC132的单泛素化表达较MOLP-2明显增加,而P-gp、MRP在耐药细胞系中表达无升高.结论 MOLP-2/R具有典型多药耐药特性,为进一步研究耐药逆转途径提供了实验基础.FANCD2蛋白单泛素化表达增强可能是MOLp-2/R细胞产生获得性耐药的主要机制之一.     

Antineoplastic efficacy of melphalan andN-(2-chloroethyl)-N-nitrosocarbamoyl-ω-lysine,in combination with diazoxide or insulin in autochthonous mammary carcinoma of the Sprague-Dawley rat

Summary The anticancer activity of melphalan andN-(2-chloroethyl)-N-nitrosocarbamoyl--lysine (CNC--Lys), was compared in the autochthonous, methylnitrosourea-induced mammary carcinoma of the Sprague-Dawley rat. In addition, the influence on the therapeutic efficacy of the combination with diazoxide, causing a mild, reversible diabetes, and with insulin was investigated. The comparison of melphalan and CNC--Lys clearly showed the superiority of melphalan. Both compounds displayed a significant tumour inhibition in their medium and the highest dosages in comparison to the untreated control. The combination with diazoxide resulted for almost all groups in an increased tumour inhibition. Only the lowest dose of CNC--Lys + diazoxide did not reduce the tumour volume significantly versus the control group. The combination with insulin, however, resulted in a loss of tumour inhibition compared to the effect of the cytotoxic drug alone, although in these groups, too, a significant decrease of tumour volumes versus controls could be observed. Mortality was within tolerable limits (<20%) through the treatment period for all experimental groups. Median lifespans were increased in all therapy groups, but no additional benefit could be observed in the combination treatment groups.Abbreviation used CNC--Lys N-(2-chloroethyl)-N-nitrosocarbamoyl--lysine     

Amidine analogues of melphalan: synthesis, cytotoxic activity, and DNA binding properties

Design, synthesis, and cytotoxic activity of amidine derivatives of melphalan are described and structure-activity relationships are discussed. Evaluation of the cytotoxicity of these compounds employing a MTT assay and inhibition of [(3)H]thymidine incorporation into DNA in both MDA-MB-231 and MCF-7 human breast cancer cells demonstrated that these compounds were more active than melphalan. Data from the ethidium displacement assay showed that these compounds were able to bind in the minor groove-binding mode in AT sequences of DNA. The cytotoxic properties of the amidine analogues of melphalan towards cultured human breast cancer cells correlate with topoisomerase II inhibitory properties but not with DNA-binding properties.     

小剂量马法兰治疗高危骨髓增生异常综合征的临床观察

目的：观察小剂量马法兰治疗高危骨髓增生异常综合征，即原始细胞过多的难治性贫血（ＲＡＥＢ）和转变中的原始细胞过多的难治性贫血（ＲＡＥＢ－Ｔ）的疗效。方法：应用马法兰２ｍｇ，隔日一次口服，治疗高危骨髓增生异常综合征１３例。结果：完全缓解５例，部分缓解２例，进步１例，总有效率６１．５％。除２例轻度骨髓抑制外，无其他毒副反应。结论：小剂量马法兰治疗高危骨髓增生异常综合征具有安全有效、服用方便、费用低廉等特     

L-Amino acid oxidase (LOX) modulation of melphalan activity against intracranial glioma

 These studies evaluated the efficacy of sequential pretreatment with L-amino acid oxidase (LOX) and LOX antiserum in the modulation of melphalan activity against intracranial glioma in athymic nude mice. LOX produced statistically significant (P<0.01) depletion of the large neutral amino acids isoleucine, leucine, methionine, phenylalanine, tyrosine, and valine in murine plasma at doses of 100 and 200 μg administered intravenously. Polyclonal anti-LOX antibody was successfully produced in mice, rabbits, and goats subsequent to immunization with LOX. Staphylococcal protein A-purified rabbit anti-LOX serum inhibited approximately 50% of LOX activity in vitro relative to control samples. This antiserum was used in vivo to inactivate LOX after it had depleted the large neutral amino acids, thereby preventing LOX-mediated catabolism of melphalan. Inoculation of three mice with rabbit anti-LOX serum after the treatment with LOX (100 μg) reduced LOX activity by 100%, 89%, and 100% at 6 h compared with reductions of 80%, 59%, and 52% over the same period in animals receiving LOX alone. In three separate studies using groups of eight to ten mice bearing intracranial human glioma xenografts, pretreatment with LOX followed by anti-LOX serum increased the antitumor activity of melphalan as compared with treatments with melphalan plus LOX, melphalan plus anti-LOX serum, or melphalan alone. Received: 4 December 1995/Accepted: 25 May 1996     

Hyperthermia modifies pharmacokinetics and tissue distribution of intraperitoneal melphalan in a rat model

Background and objectives Peritoneal surface malignancy is a common manifestation of failure of treatment for abdominal cancers. Best results of treatment have been achieved with complete cytoreduction followed by heated intraoperative chemotherapy. Melphalan is a chemotherapeutic agent that shows increased pharmacological activity with heat. But the combination of intraperitoneal administration and heat have never been tested for this drug. The purpose of this study was to evaluate the effect of hyperthermia on the pharmacokinetics and tissue distribution of intraperitoneal melphalan in a rodent model.Methods Melphalan was given by the intraperitoneal route to 20 Sprague-Dawley rats at a dose of 12 mg/kg over 90 min. Rats were randomized into two groups according to the temperature of the peritoneal perfusate: group NT received normothermic (33.5°C) melphalan; group HT received hyperthermic (42°C) melphalan. During the course of intraperitoneal chemotherapy, peritoneal fluid and blood were sampled at 5, 15, 30, 60 and 90 min. At the end of procedure, the rats were killed and tissues samples (heart, liver, ileum, jejunum, colon, omentum, and abdominal wall) were collected. Concentrations of melphalan were determined in peritoneal fluid, plasma, and tissues by high-performance liquid chromatography.Results The area under the curve (AUC) of peritoneal fluid melphalan was significantly lower in the HT group than in the NT group (P=0.001), whereas no significant difference in plasma AUC was found. AUC ratios (AUC peritoneal fluid/AUC plasma) were 12.1 for the NT group and 12.3 for the HT group. The mean time to reach the plasma peak was shorter in the HT group than in the NT group (P=0.004). The HT group exhibited increased melphalan concentrations in all intraabdominal tissues. These differences were significant for the ileum (P=0.03) and jejunum (P=0.04).Conclusion Hyperthermia affected the pharmacokinetics of intraperitoneal melphalan by decreasing the AUC of peritoneal fluid melphalan without increasing the plasma AUC. It increased intraabdominal tissue concentrations.Olivier Glehen is supported by a grant from Fondation de France.     

Hyperthermic Isolated Limb Perfusion With Low-Dose Tumor Necrosis Factor-α and Melphalan for Bulky In-Transit Melanoma Metastases

Background: Melphalan (L-PAM) hyperthermic isolated limb perfusion (HILP) is currently considered the standard treatment for patients with in-transit metastases from cutaneous melanoma. We here report on the results of L-PAM and low-dose tumor necrosis factor (TNF) HILP in patients with bulky disease.Methods: Twenty patients underwent TNF (1 mg) and L-PAM (10 mg/L) HILP. Perfusion was performed for 90 minutes, and systemic leakage was strictly monitored. Locoregional toxicity was evaluated according to Wieberdinks criteria, whereas tumor response was evaluated with physical examination and ultrasound scan with or without fine-needle aspiration of any suspected recurrence.Results: In all cases, systemic leakage was <5%. No postoperative deaths occurred, and locoregional toxicity was mild (grade 1 or 2) in 95% of patients. A complete tumor response was obtained in 14 patients (70%), and partial responses were obtained in 5 patients (25%). After a median follow-up of 18 months, six patients are alive and disease free, seven are alive with local or distant recurrence or both, and seven have died of disease.Conclusions: Low-dose TNF HILP can achieve tumor responses comparable with those reported with higher doses of cytokine. Moreover, this drug regimen is associated with acceptable local toxicity, carries a smaller risk of systemic toxicity, and incurs lower costs.     

POEMS syndrome: experience with fourteen cases

The POEMS syndrome, also known as Crow-Fukase disease, is a rare multisystem disorder, which may take several years to evolve fully. The combination of symptoms and signs is highly complex and some of the features are detected at sub-clinical level requiring high level of suspicion. The clinical data on POEMS is still evolving with only a few case reports from India. Herein, we report a series of 14 cases with POEMS syndrome at our centre over the past 8 years, which were analysed retrospectively for their clinical features, response to therapy and treatment outcome. Presence of plasma cell dyscrasia (PCD) was essential for inclusion in this study. Confirmation of PCD was done by positive "M" spike in serum and/or urine, bone marrow plasmacytosis or presence of plasmacytoma on biopsy. In addition, the diagnosis of POEMS syndrome needed the presence of at least two of the following features: polyneuropathy, organomegaly, endocrinopathy and/or skin changes. Patients were excluded from study if there was a secondary cause of polyneuropathy like amyloidosis, drugs like vincristine, nerve root or spinal cord compression. Two patients had complete form (all five features) of the syndrome, whereas 12 had incomplete form. Median age was 48 years (range 32-65). Peripheral neuropathy was seen in 13 (92.85%) cases, organomegaly 10 (71.42%), endocrinal involvement 7 (50%) and skin changes 9 (64.28%). An association with Castleman's disease and vasculitis was also noted. With different chemotherapy protocols, all treated patients (n=12), had significant symptomatic improvement with or without objective improvement at median follow up of 48 months (range 6-120). In conclusion, high level of suspicion is required to detect this rare entity.     

Japanese contributions to ocular oncology

Important studies by Japanese ocular oncologists are reviewed. These include investigations of mortality from malignant neoplasms of the eye in Japan and 17 other countries; the incidence of retinoblastoma, malignant melanoma of the eye, and lacrimal gland cancer; photodynamic therapy for retinoblastoma, using argon laser and hematoporphyrin derivatives; chemosensitivity profiles of retinoblastoma; chemo-thermotherapy, using melphalan; selective ophthalmic arterial infusion, using a balloon catheter; intravitreal injection of melphalan for the treatment of vitreous seeding of retinoblastoma; and stereotactic radiotherapy for choroidal melanoma. Received: October 22, 1999