Expression of PFKFB3 in brain glioma tissues and its effect on malignant biological behaviors of H4 cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: Toevaluatetheexpressionof6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3(PFKFB3) in malignant glioma tissues and the effects of inhibitor of PFKFB3(PFK15) on the proliferation, migration, invasion, clone formation and tumorigenesis of H4 cells. Methods: Malignant brain glioma tissues and corresponding paratumor tissues from 31 patients, who were hospitalized in Department of Neurosurgery,Ankang Hospital of Traditional Chinese Medicine during February 1, 2015 to January 31, 2016 for operative treatment, were collected for this study. Immunohistochemistry and western blotting assays were applied to detect the expression of PFKFB3 in collected tissues. PFKFB3 in H4 cells were blocked by PFK15 (1.25, 2.5, 5.0 μmol/L). The effect of PFK15 on proliferation, migration, clone formation and tumorigenesis of H4 cells were determined by MTT assay, EdU incorporation assay, wound healing assay, Transwell assay, colone formation assay and in vivo xenograft bearing nude mice model respectively. Results: Positive expression rate of PFKFB3 was significantly higher in malignant glioma tissues compared with normal adjacent tissues[(80.60±8.98)% vs (41.57±10.16)%, P<0.05]. The results of MTT assay and EdU incorporation assay indicated that PEK15 significantly inhibited the proliferation of H4 cells in a concentration dependent manner. The migration, invasion and clone formation activity of H4 cells were significantly reduced by treatment with PFK15 (all P<0.05). In tumor bearing nude mice, the tumor volume of mice treated with PFK15 was significantly smaller than that of mice from control group ([254.15±154.25] vs [801.52±224.25] mm3, P<0.05). Conclusion: PFKFB3 was highly expressed in malignant glioma tissues. Blocking of PFKFB3 by PFK15 significantly reduced the malignant biological behaviors and tumorigenesis of H4 cells in vitro and in vivo, which may serve as a promising target for the treatment of malignant gliomas.

Malignant Glioma with Neuronal Marker Expression : A Clinicopathological Study of 18 Cases

OBJECTIVE: Malignant gliomas with neuronal marker expression (MGwNM) are rare and poorly characterized. Increasingly diverse types of MGwNM have been described and these reported cases underscore the dilemmas in the classification and diagnosis of those tumors. The aim of this study is to provide additional insights into MGwNM and present the clinicopathological features of 18 patients. METHODS: We reviewed the medical records of 18 patients diagnosed as MGwNM at our institute between January 2006 and December 2012. Macroscopic total resection was performed in 11 patients (61%). We evaluated the methylation status of O6-methylguanine-DNA methyltransferase (MGMT) and expression of isocitrate dehydrogenase 1 (IDH-1) in all cases, and deletions of 1p and 19q in available cases. RESULTS: The estimated median overall survival was 21.2 months. The median progression-free survival was 6.3 months. Six patients (33%) had MGMT methylation but IDH1 mutation was found in only one patient (6%). Gene analysis for 1p19q performed in nine patients revealed no deletion in six, 19q deletion only in two, and 1p deletion only in one. The extent of resection was significantly correlated with progression free survival on both univariate analysis and multivariate analysis (p=0.002 and p=0.013, respectively). CONCLUSION: In this study, the overall survival of MGwNM was not superior to glioblastoma. The extent of resection has a significant prognostic impact on progression-free survival. Further studies of the prognostic factors related to chemo-radio therapy, similar to studies with glioblastoma, are mandatory to improve survival.

Mortalidad por tumores cerebrales malignos durante los primeros 30 días de la cirugía / Mortality from malignant brain tumors within the first 30 days after surgery

Introducción: las complicaciones neurológicas en la cirugía de los gliomas de alto grado están relacionadas con problemas en localización y exposición, extensión de la resección y la manera en que se manipulan los tejidos. Hay factores secundarios que contribuyen a la evolución desfavorable y la muerte precoz de un paciente. Objetivos: establecer la relación que existe entre la mortalidad en los primeros 30 días y diferentes factores de riesgo e identificar causas de muerte. Métodos: fueron operados 131 pacientes en el Servicio de nNeurocirugía del Hospital Calixto García desde enero de 2005 a enero de 2010, con el diagnóstico de gliomas de alto grado. De ellos, 14 fallecieron en los primeros 30 días. Variables principales utilizadas: edad, localización tumoral, grado de resección quirúrgica y causas de la muerte. Se aplicó la prueba chi cuadrado de independencia con un nivel de significación de 0,05 para evaluar relación entre variables. Resultados: la mortalidad perioperatoria fue del 10,7 por ciento. El riesgo fue mayor en el sexo masculino; discretamente mayor en los mayores de 60 años. No estuvo relacionado con la localización del tumor. Operar con signos de enclavamiento cerebral, incrementó al máximo la probabilidad de fallecer en el primer mes. La resección parcial en relación con la total y con la biopsia mostró mayor probabilidad de mortalidad. No hubo muerte transoperatoria. Conclusiones: los factores que influyeron principalmente en la evolución desfavorable fueron: la baja puntuación en la escala de Karnofskyy una resección limitada, igual o menor que 50 por ciento en una craneotomía(AU)

Introduction: The neurological complications found in the high gradegliomas surgery are primarily related with location, exposure and extension of the resection and with the way of handling tissues. There are secondary factors that contribute to the unfavorable progression and even the early death of patients that suffers this disease. Objectives: To set the relationship between mortality within the first 30 days after surgery and the different risk factors, and to identify the causes of death. Methods: One hundred and thirty one patients diagnosed with high gradegliomas were operated on in the neurosurgery service of Calixto García hospital from January 2005 through January 2010. Fourteen of them died within the first thirty days. The study variables were age, tumor location, degree of surgical resection and causes of death. Chi-square test of independence with significance level of 0.05 was applied to evaluate the association among the variables. Results: The perioperative mortality rate was 10.7 percent. The risk of death was higher in males; slightly higher in those over 60 years and unrelated to the tumor location. If patients are operated on with signs of brain herniation, they are more likely to die in the first month. Partial compared to total resection and to biopsy showed higher mortality probabilities. There was no intraoperative death. Conclusions: The low scoring in the Karnofsky Scale, and a partial resection equal or smaller than 50 percent in a craniotomy were the influential factors in an unfavorable outcome of disease(AU)

Treatment of recurrent glioblastoma with intra-arterial BCNU [1, 3-bis (2-chloroethyl)-1-nitrosourea] / Tratamento do glioblastoma recorrente com BCNU [1, 3-bis (2-chloroethyl)-1-nitrosourea] intra arterial

Contemporary therapies for patients with glioblastomas remain marginally efficient, and recurrence following surgery, radiation therapy and adjuvant chemotherapy is practically universal. The major obstacles to the successful use of chemotherapy for CNS tumors are the drug delivery to the tumor site and the infusion of chemotherapeutic agents directly into the arterial supply of a tumor. The latter could provide a pharmacokinetic advantage by enhancing drug delivery to the tumor. Sixteen patients with recurrent unilateral glioblastomas treated with intra-arterial BCNU were evaluated retrospectively. During the infusion, eleven patients referred pain in the ipsilateral eye, five patients were nauseated, three reported headache, one patient presented mental confusion, while two presented focal signs. There were two deaths during the course of therapy. Four patients achieved temporary clinical improvement, seven showed disease stability, and three presented clinical deterioration. The median total survival time was 87.9 weeks. Unilateral vision loss and focal signs were observed as delayed complications of this treatment. This study has confirmed previous reports indicating that arterial chemotherapy is clearly not curative, and presents serious toxicity. Only through a randomized prospective study performed in a large series of patients can the questions concerning survival period increment be answered properly.

Os tratamentos atuais para pacientes com glioblastoma permanecem pouco eficientes e a recorrência, acompanhando cirurgia, radioterapia e quimioterapia, é a regra geral. O maior obstáculo para o sucesso da quimioterapia para os tumores do SNC é a disponibilização da droga no sitio do tumor sendo que a infusão do agente quimioterápico diretamente na trama arterial da lesão pode proporcionar vantagens por maior liberação da substância diretamente no tumor. Estudamos retrospectivamente dezesseis pacientes com glioblastomas recorrentes, unilaterais, que foram tratados com BCNU intra-arterial; durante a infusão, onze pacientes sentiram dor no olho ipsilateral, cinco ficaram nauseados, três queixaram-se de cefaléia, um apresentou confusão mental e dois apresentaram sinais focais. Ocorreram duas mortes durante a terapia. Quatro pacientes apresentaram melhora clinica temporária, sete apresentaram estabilização e três apresentaram deterioração. A média de sobrevida total foi de 87,9 semanas. Perda da visão unilateral e sinais focais foram complicações tardias. Este estudo confirmou trabalhos anteriores indicando que a quimioterapia intra-arterial claramente não é curativa, séria toxicidade pode ocorrer e somente um estudo prospectivo e randomizado, realizado em uma serie maior de pacientes, poderá responder questões sobre o aumento do tempo de sobrevida de forma adequada.

Effect of High-Dose Tamoxifen on Malignant Gliomas

In vitro studies have shown that the nonsteroidal antiestrogen tamoxifen can suppress deoxyribonucleic acid(DNA) synthesis and cell proliferation in cultured human gliomas. This growth suppression is independent on its antiestrogenic properties. Tamoxifen may act through the inhibition of the enzyme protein kinase C(PKC), which transduces mitogenic signals from the cell surface to the nucleus. In order to evaluate the therapeutic response and side effect of high-dose tamoxifen, we performed a clinical study of 28 patients with malignant gliomas who were treated with high-dose tamoxifen in our hospital between February 1991 and January 1993. An effect was defined as a statistically improved survival times/rates. In patients who were assigned to receive high-dose tamoxifen, it was first administered at standard antiestrogen doses(20mg orally bid/day) to observe for any side effect and if tolerated, the dose was increased weekly to achieve target doses(100mg orally bid/day) over a 1 month period. We compared the survival times/rates between anaplastic astrocytomas and glioblastoma mutiformes. Although the median survival time was slightly longer in anaplastic astrocytomas than that of glioblastoma multiformes, there was no statistical difference of survival curves between two groups at the p=0.05 level. We also examined the survival times/rates of malignant gliomas according to treatment modalities(radiotherapy alone, radiotherapy plus ACNU, and radiotherapy plus tamoxifen). Although the survival rate and time were slightly higher in radiotherapy plus tamoxifen group than those of another treatment groups, we could not find the statistical significance of survival curves between three treatment groups(p>0.05). High-dose oral tamoxifen appeared to be well tolerated in most patients. Five patients developed anorexia following dose escalation of tamoxifen. Another complications were amenorrhea, nausea/vomiting, and constipation. There were no changes in hematological studies that could be attributed to tamoxifen. We think that high-dose tamoxifen cah be administered safely to malignant gliomas patients. Our results were not impressive. We conclude that the definition of the true efficacy of high-dose tamoxifen in patients harboring malignant gliomas is not possible from this limited study, and a further large scale, randomized trial of this agent is necessary.

Effect of ACNU Chemotherapy on Malignant Glioma

In order to determine if there was an enhancing therapeutic effect of ACNU(1-4-amino-2-methyl-5-pyrimidinyl)-methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride, nimustine chloride given in addition to radiotherapy, we performed a randomized clinical study of irradiation alone and combination of irradiation with ACNU in the treatment of malignant gliomas. Thirty-seven patients who were treated in our hospital from August 1990 to September 1992 were included in this study. An effect was defined as a statistically improved survival times. Radiotherpy with a total dose of 5000 to 6500 rads was applied to the whole brain and to a generous field surrounding the tumor. Patients who were assigned to receive chemotherapy were given ACNU intravenously at a dose of 1-2mg/kg. The survival rates of patients with anaplastic astrocytoma and glioblastoma multiforme at 18 months after the surgery were 0% and 37% for radiotherapy alone, and 66.7% and 40.1% for radiotherapy plus ACNU, respectively. The median survival times of patients with anaplastic astrocytoma and glioblastoma multiforme were 14 and 15 months for radiotherapy alone, and 19 and 16 months for radiotherapy plus ACNU, respectively. The survival rates of patients with malignant gliomas(anaplastic astrocytoma plus glioblastoma multiforme) at 18 months were 5.5% for radiotherapy alone and 45.4% for radiotherapy plus ACNU, and the median survival times were 15 and 16 months, respectively, Althouh the survival rate of patients with malignant gliomas at initial 6 months was much higher in radiotherapy plus ACNU than in radiotherapy alone, the differences between survival curves were not significant at the p=0.05 level. This study demonstrated that, although the use of ACNU during radiotherapy suppressed malignant gliomas more than radiotherapy alone, the survival time was not extended significantly. It is necessary to continue to search for an effective chemotherapapeutic regimen to prolong survival of patients with malignant gliomas.