Cross-resistance pattern in brain tumour cells resistant to antitumour chloroethylnitrosoureas.

We tested acquired resistance and cross-resistance of brain tumour cells after repeated treatments of antitumour agents including chloroethylnitrosoureas (CENUs) in clinical use for brain tumour chemotherapy. Within ten repeated 2-day incubation periods with either 1-(4-amino-2-methyl-5-pyrimidinyl) methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride (ACNU) or methyl-6-[3-(2-chloroethyl)-3-nitrosoureido]-6-deoxy-alpha-D-glucopyr anoside (MCNU), brain tumour cells (9L) developed high degrees of resistance to these drugs, as evidenced by about 3- and 6-fold increases, respectively, at the 10% survival dose (SD10). The resistance has unchanged with time after termination of challenging treatments. Repeated challenges with bleomycin (BLM), cis-diaminedichloroplatinum (II) (CDDP), and methotrexate (MTX) did not develop a significant resistance. ACNU-resistant (9L/ACNU) cells showed complete cross-resistance to MCNU, and MCNU-resistant (9L/MCNU) cells to ACNU. Drug sensitivity to other DNA-damaging agents (BLM, NCS, CDDP, etoposide) than CENUs fluctuated to a lesser extent among 9L parent, 9L/ACNU, and 9L/MCNU cells. These data suggest clinical disadvantage of prolonged adjuvant CENU chemotherapy and advantage of combined CENU chemotherapy with other agents than CENUs in brain tumours.     

DNA lability induced by nimustine and ramustine in rat glioma cells.

The DNA labile sites induced by two nitrosoureas, nimustine (ACNU) and ramustine (MCNU) synthesised in Japan, have been examined in highly reiterated DNA sequences of rat glioma cells. Reiterated fragments of 167 and 203 base pairs (bp), obtained after Hind III and Hae III restriction endonuclease digestion of rat glioma cells DNA, were used as target DNA sequences to determine the labile sites. In vitro reaction with ACNU and MCNU resulted in scission products corresponding to the locations of guanine. Subsequent piperidine hydrolysis produced more frequent breaks of the phosphodiester bonds at guanine positions, thus forming alkali-labile sites.     

Intrathecal distribution of ACNU by various modes of its administration analyzed by HPLC and autoradiography]

Various modes of administration of ACNU (nimustine hydrochloride) were tried to make clear which mode is the best method to obtain intrathecal diffuse distribution of ACNU to match the condition of killing of glioma cells (10 micrograms/ml; greater than 30 min.). Tried modes of administration included 1)bolus injection into ventricular cavity, 2)bolus injection into cisterna magna, 3)bolus injection into lumbar subarachnoid space, 4)ventriculo-lumbar perfusion, 5)chiasmatic cistern-lumbar perfusion. Used dose of ACNU was 5 mg/body for all modes of administration. ACNU level in CSF was measured by HPLC method specially developed by authors. To make clear intrathecal distribution of ACNU, autoradiography using 14C-ethylene-ACNU was studied after administration of 10 muCi/Kg of radioactive ACNU. The images were studied by image analyzer system (BAS-2,000 system developed by Fuji Film Co. Ltd). Among the modes of administration tried, ventriculo-lumbar perfusion method gave the best results in terms of lumbar, ventricular, cisterna magna, and basal cistern distribution of ACNU to match the cell kill condition experimentally ascertained. Although, bolus injection of ACNU into cisterna magna gave sufficient amount of ACNU in lumbar region, the initial level of ACNU was too high in cisterna magna, and administration of ACNU once a week for three times in a canine cisterna magna resulted in considerable deterioration of brain stem and basal structure. In addition to it, the level of ACNU in ventricular cavity was not detectable. Lumbar bolus injection resulted in also too much ACNU accumulation at the injected lumbar area, and at the cisterna magna region, ACNU was not detectable.(ABSTRACT TRUNCATED AT 250 WORDS)     

Neoplastic meningitis from breast carcinoma with complete response to liposomal cytarabine: case report

Neoplastic meningitis from breast cancer often leads to a progressive neurologic deterioration followed by fatal outcome. The therapy is based on the administration of high dose systemic chemotherapy with drugs able to pass through the blood-brain barrier, such as methotrexate (MTX) and cytarabine, cranial or craniospinal irradiation, and intrathecal (IT) administration of MTX and/or cytarabine. However, these approaches only have modest efficacy and are associated with side effects for the patients. A depot formulation of liposomal cytarabine (DepoCyte^®) has proven to be useful in clinical trials. We describe the case of a woman with a diagnosis of leptomeningeal carcinomatosis from breast carcinoma who presented cerebrospinal fluid normalization and prolonged complete MRI response to intrathecal chemotherapy with liposomal cytarabine (DepoCyte).     

Chemotherapy for progressive pilocytic astrocytomas in the chiasmo-hypothalamic regions

Over the past 25 years, we have treated 17 patients with chiasmo-hypothalamic astrocytomas. Before 1988, the initial treatments consisted of surgery and/or radiotherapy, while since 1989, 4 children (1 male, 3 females, aged 3–8 years) were treated primarily with chemotherapy. None of them was associated with neurofibromatosis. After a biopsy of the tumor, the intraveneous administration of ranimustine (MCNU; 30–86 mg/m²) and/or nimustine (ACNU; 30.3–64.1 mg/m²) was given without radiation therapy. Chemotherapy was usually given as an out-patient, with a total of 5–13 courses. The total doses of MCNU and ACNU administered ranged from 150 to 570 mg and from 64.8 mg to 100 mg, respectively. After chemotherapy 2 patients showed clinical improvement and tumor regression on neuro-imaging, while one patient showed clinical improvement and tumor size stabilization on neuro-imaging. The remaining one child, however, showed a clinical worsening and tumor progression on neuro-imaging studies. He was thus treated with a second chemotherapy regimen with carboplatin and etoposide, which brought about tumor regression. The acute and subacute toxicity of chemotherapy was mild. All patients are now leading almost normal lives with a median of 43 months after diagnosis. Although a longer and more careful clinical observation is required, the quthors conclude that chemotherapy with MCNU and/or ACNU may benefit patients with unresectable pilocytic astrocytoma requiring treatment. The advantages of this therapy include its mild side effects and the lack of any hospitalization in most patients. It may also delay the need for radiation therapy, which can have a deleterious effect on the young developing brain.     

脑胶质瘤化疗敏感性实验研究

目的探讨脑胶质瘤细胞体外对化疗药物的敏感性，为临床有效治疗胶质瘤提供实验依据。方法采用四甲基偶氮唑盐微量酶反应比色法（MTT法）检测50例原代培养胶质瘤细胞在体外对替尼泊甙（VM-26）、卡氮芥（BCNU）、顺铂（CDDP）、长春新碱（VCR）、甲氨蝶呤（MTX）5种化疗药物单一或联合化疗的敏感性。结果42例获得药敏结果，成功率达84％。42例胶质瘤细胞对单一化疗药物的敏感性从高到低依次为VM-26〉BCNU〉CDDP〉VCR〉MTX。两种联合化疗方案（BCNU＋VM-26＋CDDP）和（BCNU＋VCR＋MTX）对胶质瘤细胞的敏感性明显优于单一用药（P〈0．01），且对绝大多数胶质瘤细胞增殖抑制率较高。结论两种联合化疗方案（BCNU＋VM-26＋CDDP）和（BCNU＋VCR＋MTX）可能对胶质瘤的化疗效果更好。     

Adult medulloblastoma

Adult medulloblastoma is a rare tumor with few retrospective studies published so far. The role of adjuvant chemotherapy or chemotherapy at relapse is unclear. This study reports therapy and outcome in all adult ( 16 years old) medulloblastoma (n = 34) and supratentorial primitive neuroectodermal tumor (PNET) patients (n = 2) treated in 2 neuro–oncological centers between 1976 and 2002. The median age was 24.5 years (range 16–76). After resection, 16 patients were treated with craniospinal radiotherapy alone, 20 patients also received adjuvant chemotherapy (8 vincristine, CCNU, cisplatin; 7 methotrexate alone or methotrexate/vincristine–based polychemotherapy; 5 other protocols). Median survival in the whole cohort was 126 months (2+ – 200+ months). Five–year and 10–year survival rates were 79 % and 56%. Adjuvant chemotherapy was associated with a non–significant trend to prolonged survival (relative risk (RR) 1.89; p = 0.068). The median progression–free survival (PFS) after primary therapy was 83 months. At relapse, 10 of 12 evaluable patients achieved a complete response upon second–line therapy. The median survival times from first (n = 17) and second relapse (n = 9) were 21 months (0–67+ months; 5/17 without second relapse) and 20 months (1–29 months). Cox regression analysis revealed the infiltration of the floor of the 4^th ventricle at diagnosis as the only therapy–independent prognostic factor (RR 0.48; p = 0.03). In conclusion, adjuvant chemotherapy may prolong survival in adult medulloblastoma patients. Moreover, second–line therapy may be beneficial for these patients. As in pediatric medulloblastoma patients, primary infiltration of the floor of the 4^th ventricle indicates a poor prognosis.This publication is dedicated to the late Joachim Kühl, MD, whose life was dedicated to improve therapy for medulloblastoma patients.*Drs. Herrlinger and Steinbrecher contributed equally to this work.     

Intrathecal chemotherapy for leptomeningeal dissemination of medulloblastoma

Cerebrospinal fluid dissemination of medulloblastoma occurs despite adequate systemic chemotherapy, and unless it is present at the time of initial diagnosis, occurs late in the course of the disease. Intrathecal chemotherapy with an implanted Ommaya reservoir can produce short-term benefit. We report here our protocol for and the results of intrathecal chemotherapy for leptomeningeal dissemination of medulloblastoma.     

Intrathecal methotrexate affects cognitive function in children with medulloblastoma

BACKGROUND: Cognitive impairment occurs after malignant brain tumor treatment in children, following brain radiotherapy and systemic and intrathecal chemotherapy. OBJECTIVES: 1) To compare two groups of children who underwent surgery for cerebellar medulloblastoma with their cousins and siblings, assessing intelligence, executive function, attention, visual perception, and short-term memory. Both groups were treated with the same combined radiotherapy-chemotherapy, but differed in that only one group received intrathecal methotrexate (MTX+). 2) To relate these measures to MRI findings (leukomalacia). RESULTS: The two groups performed worse than their control subjects in all tests. The MTX+ group younger than 10 years performed significantly worse in all tests, particularly executive ones. The group older than 10 years performed significantly worse only in short-term memory. Younger patients without MTX performed significantly worse than controls only in some neuropsychological measures; there were no differences between older patients and control subjects. Only in the MTX+ group was there a direct correlation between extent of leukomalacia and performance in some tests. CONCLUSIONS: The administration of intrathecal methotrexate to children with medulloblastoma worsens the cognitive deficits induced by chemotherapy and radiotherapy. The use of intrathecal methotrexate in the treatment of medulloblastoma and other malignancies should be reassessed.     

恶性脑肿瘤的超选择性脑动脉灌注治疗

目的:对恶性脑胶质细胞瘤术后残留的肿瘤组织,用化疗方法将其杀灭和抑制生长,以延长病人的生存期和改善生存质量;并比较嘧啶亚硝脲(ACNU)加氨甲喋呤(MTX)(A组)以及三尖杉酯碱加MTX(B组)的疗效。方法:将ACNU2～3mg/kg,三尖杉酯碱0.1～0.5mg/kg,MTX0.1～0.2mg/kg按A、B两组方案,采用超选择性脑动脉内联合灌注化疗,每例1～5次不等。结果:A组12例中CR1例,PR3例,NC7例,PD1例;B组12例中CR1例,PR2例,NC7例,PD2例。A组和B组对恶性脑胶质瘤的有效率分别为33.33％和25％,化疗后肿瘤体积缩小率分别为48.32％和46.21％,各组治疗前后肿瘤体积变化均有显著性差异,但两组疗效对比无明显差异。结论:恶性脑胶质细胞瘤术后辅以超选择性脑血管内化疗,能抑制肿瘤生长,ACNU加MTX与三尖杉酯碱加MTX的联合化疗方案合理,毒副作用小。     

Primary CNS lymphoma: combined treatment with chemotherapy and radiotherapy

Primary central nervous system lymphoma (PCNSL), an uncommon tumor, is occurring with increasing frequency. Conventional therapy with corticosteroids and cranial radiotherapy (RT) usually gives a dramatic initial response, but median survival is only 10 to 18 months. Chemotherapy is more successful in comparable systemic lymphoma and has been employed for PCNSL at relapse, causing remission but not cure. Between June 1985 and June 1988, we prospectively staged 32 patients with PCNSL at Memorial Sloan-Kettering Cancer Center and treated 28 on a new protocol that combined chemotherapy and radiotherapy at diagnosis. None had occult systemic lymphoma, but 19% had ocular and 69% had definite or probable leptomeningeal lymphoma. There were no complications in 19 stereotactic biopsies, but 4/10 patients who had a complete resection suffered a severe postoperative deficit. Four patients received RT alone, and 28 received chemotherapy and cranial RT, 17 of whom (group A) received a combination regimen using pre-RT systemic (1 g/m2) and intra-Ommaya methotrexate (MTX), 4,000 cGy whole-brain RT with a 1,440 cGy boost, and 2 courses of post-RT high-dose cytosine arabinoside; 5 other patients received an identical regimen but with a decreased dose of MTX (200 mg/m2). Sixty-three percent of assessable patients had a response to MTX independent of corticosteroid and prior to RT. Eighteen of 26 (69%) assessable patients who received combined therapy are alive with a median follow-up of 25.4 months. Twelve of 16 (75%) assessable group A patients are alive in the same period. Chemotherapy-related toxicity was minimal, and no late toxicities have occurred to date.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chemotherapy for malignant brain tumors of childhood

During the past 3 decades, chemotherapeutic agents for the treatment of pediatric brain tumors have been extensively evaluated in a myriad of schedules, doses, and combinations. Remarkable advances in outcome have been achieved for certain children, notably those with medulloblastoma, but with a high cost to quality of life. In addition, the success achieved for medulloblastoma is offset by lack of progress for high-grade glioma. Despite intensive investigation, no single chemotherapeutic regimen stands out for children with high-grade glioma, with most succumbing to their disease. Further treatment intensification using conventional nonspecific chemotherapy is more likely to result in additional toxicity without major advances in survival. Genome-wide analysis using microarray technology has contributed significantly to our understanding of tumor biology, shifting the focus onto novel agents that target molecular changes crucial for tumor proliferation or survival. These selective agents are likely to be less toxic to normal cells and more effective.     

Acute fever and delayed leukoencephalopathy following low dose intraventricular methotrexate.

Nine out of 14 patients treated with intraventricular methotrexate (MTX) for meningeal carcinomatosis from breast carcinoma and surviving more than 4 months developed disseminated necrotising leukoencephalopathy (DNL). All four patients who had received both intraventricular MTX and whole brain radiotherapy developed DNL. Five of the six patients who experienced an acute febrile reaction with mild encephalopathic signs following intraventricular administration of MTX developed DNL after a mean time of 5 months and a low mean dose of 44 mg MTX. DNL was also noted in two patients without a previous febrile reaction or whole brain radiotherapy, following prolonged intraventricular MTX therapy after a mean time of 19.5 months and a mean dose of 147 mg MTX. These findings confirm the hazards of (1) high cumulative doses of intrathecal MTX and (2) combined intrathecal chemotherapy and whole brain radiotherapy. This study also suggests a possible relationship between an early and transient febrile reaction during intraventricular administration of MTX and the development of DNL.     

Babinski and the diagnosis of amyotrophic lateral sclerosis

Cytological evaluation of cerebrospinal fluid (CSF) is an important means of following response to intracavitary chemotherapy for leptomeningeal malignancy. We studied the feasibility of quantitative cytological evaluation by retrospective analysis of serial CSF specimens from 7 patients receiving phase I intracavitary chemotherapy for leptomeningeal malignancy who had persistent malignant cytology. Three to 34 CSF specimens per patient obtained over a 3- to 48-week period were reviewed. Significant (five- to 10-fold or greater) reductions in numbers of malignant cells in CSF during treatment could be identified in specimens otherwise diagnosed as positive. Quantitative CSF cytological evaluation is neither overly time consuming nor tedious to perform and may provide useful clinical information.     

Prognostic significance of the immunohistochemical expression of O6‐methylguanine‐DNA methyltransferase,P‐glycoprotein,and multidrug resistance protein‐1 in glioblastomas

We studied the expression of O⁶‐methylguanine‐DNA methyltransferase (O⁶‐MGMT), P‐glycoprotein (Pgp), and multidrug resistance protein‐1 (MRP‐1) in 23 glioblastomas using RT‐PCR, methylation‐specific PCR, and immunohistochemistry, and analyzed their association with overall patient survival. Univariate analysis of collected data demonstrated that the expressions of O⁶‐MGMT and MRP‐1 detected by immunohistochemistry, in addition to the consistent factors, including preoperative Karnofsky performance scale (KPS), radical surgery, and tumor location and extension, were significant prognostic factors for the overall survival (OS) of patients with glioblastoma, who received nimustine (ACNU)‐based chemotherapy in association with surgery and radiotherapy. Among them, following multivariate analysis, preoperative KPS, radical surgery, tumor location, and the expression of O⁶‐MGMT remained as significant prognostic factors. These findings suggest that immunohistochemical analysis of O⁶‐MGMT in patients with glioblastoma can be a useful method to predict the effects of chemotherapy and identify alternative chemotherapeutic regimens for O⁶‐MGMT‐positive patients.     

Evaluation of ACNU alone and combined with tegafur as additions to radiotherapy of the treatment of malignant gliomas--a cooperative clinical trial

Controlled, prospective, randomized studies were performed to evaluate the effects of 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3- (2-chloroethyl)-3-nitrosourea hydrochloride (ACNU) and ACNU plus tegafur as additions to radiotherapy for the treatment of malignant gliomas. In the first trial, 105 patients with glioblastoma or anaplastic astrocytoma were randomly divided into two groups after surgery and received radiotherapy (RT, 40 to 60 Gy to the whole brain), or radiotherapy plus concomitant chemotherapy with ACNU (100 mg/m2 on day 1 and 42). Effects of the treatment were compared in 82 evaluable patients from results of CT scans taken before and one month after the completion of radiotherapy. The regression rates more than 50% of the tumor size were observed in 15.0% of patients treated with RT alone and in 47.6% of patients treated with RT plus ACNU. The difference was statistically significant (p less than 0.005). In the second trial, 87 patients were randomly divided into two groups and received RT plus ACNU, or RT plus combined chemotherapy with ACNU and tegafur (400 mg/m2, daily for 8 weeks). Sixty-nine patients were within the valid study group. The regression rates more than 50% of the tumor size were observed in 34.2% of patients treated with RT plus ACNU: and in 41.2% treated with RT, ACNU plus tegafur. No statistical difference was noted in the response rate between the groups. These results indicate that ACNU is an effective agent in conjunction with radiotherapy for patients with malignant gliomas, and that tegafur does not enhance the effectiveness of ACNU.     

脑肿瘤超选择性脑动脉灌注化疗的临床分析

目的:研究超选择性脑动脉插管灌注嘧啶亚硝脲(ACNU)或卡氮芥(BCNU)治疗恶性脑肿瘤的近期疗效及其并发症的防治。方法:对67例经组织学确诊的恶性脑肿瘤采用超选择性脑动脉插管灌注嘧啶亚硝脲或卡氮芥化疗,并对照化疗前后的影像学表现,评价该化疗对恶性脑肿瘤的疗效。结果:本组67例病人治疗后完全缓解10例(14.9％),部分缓解43例(64.2％),稳定11例(16.4％),恶化3例(4.5％)。对部分病例行免疫组化和神经病理研究,证实超选择性脑动脉灌注化疗可诱导肿瘤的凋亡。超选择性脑动脉插管灌注化疗可明显降低常规化疗的并发症和毒性反应,并发症和毒性反应的发生率明显低于文献报道。结论:超选择性脑动脉插管灌注化疗治疗恶性脑肿瘤可明显降低药物毒性,近期疗效良好。     

Primary central nervous system lymphoma: a retrospective study

Eleven patients with primary central nervous system lymphoma (PCNSL) proven with biopsy between January 1991 and September 1995 were analysed according to the location of lesions, pathological studies, treatment and outcome. The ages of these patients ranged from 25 to 74 years with an average of 51.4 years. The male to female sex ratio was 6 to 5. The most frequent sites of lesions were frontal lobes. One patient had only leptomeningeal involvement. All intracranial PCNSL in this report arose from B cells; the leptomeningeal PCNSL case was the only one which arose from T cells. All patients received chemotherapy with or without radiotherapy, except two patients who did not receive any treatment. The results of treatment were generally poor. The survival time of patients who died ranged from 2.5 months to 45 months with a median of about 12.5 months. If a combined treatment is to be considered, the possibility of complications such as leucoencephalopathy should be taken in account.     

脑胶质瘤组织培养化疗药物敏感性的研究

目的建立脑胶质瘤的组织培养药敏检测法，用于筛选化疗药物及指导临床个体化化疗。方法应用组织培养药敏检测法对69例人脑胶质瘤标本进行化疗药物敏感性测定，所测药物为脑肿瘤常用化疗药物顺铂（DDP）、长春新碱（VCR）、替尼泊甙（VM26）、尼莫司汀（ACNU）、替莫唑胺（TMZ）及DDP＋VM26组合。比较6组化疗药物体外抗肿瘤作用的差异，并分析药物敏感性与病人临床资料之间的相关性。结果DDP＋VM26组合用药的敏感性与VM26单独用药比较，差异无统计学意义．但明显优于其他4种单药。病人性别、年龄、肿瘤初发或复发均对药物敏感性无显著性影响；除组合用药DDP＋VM26外，其他药物的敏感性与病理分级无显著相关性。结论胶质瘤标本不同个体对同一药物及同一个体对不同药物的敏感性差异很大，脑胶质瘤化疗应优先选择以VM26为主的联合用药。     

Relationship between expression of O6-methylguanine-DNA methyltransferase,glutathione-S-transferase pi in glioblastoma and the survival of the patients treated with nimustine hydrochloride: an immunohistochemical analysis

Drug resistance is one of the important factors that determine tumor response to chemotherapy. Several candidates for resistance to various chemotherapeutic agents have been elucidated. O6-methylguanine-DNA methyltransferase (MGMT) removes methylation damage induced by nitrosourea from the O6 position of DNA guanines before cell injury. Glutathione-S-transferase (GST) pi is also involved in nitrosourea resistance. We examined the expression of MGMT and GST pi in 18 glioblastomas (GBM) using immunohistochemistry and compared the results with patients' survival after administration of 1-(4-amino-2-methyl-5-pyrimidinyl) methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride (ACNU)-based chemotherapy. According to the Kaplan-Meier's method, although median progression free survival (PFS) of eight patients whose tumors retained high MGMT (3+ approximately 2+), and 10 patients whose tumors showed low MGMT expression (1+ approximately 0) were nine and 15 months, respectively (p = 0.09), median overall survival (OS) of the two groups were 12 and 22 months, respectively, which were significantly different (p = 0.01). GST pi expression in GBM was not a prognostic factor. It is suggested that GBM with strong staining of MGMT activity may show more resistance to ACNU-based chemotherapy compared to that with low MGMT. The simple immunohistochemical analysis of MGMT in GBM can be a useful method to determine whether ACNU or another treatment regimen should be recommended.