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Objective To explore the role of breast/ovarian cancer susceptibility gene 1 associated protein 1(BAP1)in the occurrence and progression of human malignant glioma and the feasibility of BAP1 as a clinical diagnostic marker for malignant glioma.Methods The differential expression of BAP1 in normal and glioma tissue was analyzed based on the GSE4290 and GSE90598 sub-datasets from the gene expression omnibus(GEO)database.Receiver operating characteristic(ROC)curve analysis was conducted to assess the early diagnostic value of BAP1 for malignant glioma.Primary lesion tissues from 28 nonpaired malignant glioma patients and non-tumor brain tissues removed by internal decompression surgery in 5 patients with traumatic brain injury collected independently were collected,and the expression levels of BAP1 were measured using quantitative real-time polymerase chain reaction(qRT-PCR).Specific small interfering RNAs(siRNAs)targeting BAP1 were transiently transfected into U251 cells to further evaluate their interference efficiency.Flow cytometry was employed to analyze changes in the cell cycle and apoptosis of U251 cells with BAP1 knockdown.Results The results of bioinformatics showed that the expression of BAP1 in malignant glioma tissues was lower than that in normal brain tissues(GSE 4290:1 209±18.49 vs 1 476±53.90,GSE 90598:5.19±0.10 vs 5.65±0.21),and the differences were significant(t=5.115,2.267,all P<0.05).ROC curve showed that BAP1 could efficiently differentiate malignant glioma tissue from normal brain tissue(GSE4290:AUC=0.78,GSE90598:AUC=0.75,all P<0.05).The expression level of BAP1 in primary malignant glioma tissue was lower than that in normal brain tissue(0.27±0.04 vs 1.06±0.07),and the difference was significant(t=10.22,P<0.001).After down-regulating the expression of BAP1 in U251 cells,the proportion of S phase cells increased from 17.59%to 27.21%(siBAP1-1)and 25.79%(siBAP1-2),respectively,and the differences were significant(t=6.576,6.642,all P<0.01).However,the apoptosis levels decreased from 10.17%to 2.70%(siBAP-1)and 3.00%(siBAP-2),respectively,and the differences were significant(t=10.31,9.428,all P<0.01).Conclusion Histone H2A deubiquitinase BAP1 could exert the function of tumor suppressor genes by inhibiting rapid cell cycle progression and promoting apoptosis in malignant glioma,and could serve as a potential clinical diagnostic biomarker for malignant glioma.
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Malignant glioma is the most common primary tumor of the central nervous system in adults, and it exhibits complex biological behavior and high malignancy. However, current therapies for malignant glioma are limited. Despite standard radiation therapy, maximal safe surgical resection, and combined radiochemotherapy, clinical outcomes remain dismal. Patients diagnosed with GBM only have a median survival of approximately 15 months. Several molecular targets and immunotherapies have recently emerged, and the field has made great progress, particularly with respect to oncolytic virus, which has received extensive attention among researchers because of its unique targeting, security, and antitumor effects. Oncolytic virus therapy has demonstrated considerable efficacy in basic and clinical research of various cancers. Furthermore, relevant investigations of malignant glioma are being conducted for its better understanding. Here, we review the recent clinical research progress in oncolytic virus therapy for treating malignant glioma.
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Objective: To observe the expressions of SENP1, SENP2 and SENP6 proteins in human malignant glioma tissue and cells∗ and to elucidate the their effects in the development of malignant glioma. Methods: The samples of normal human brain tissue and malignant glioma tissue were obtained and used as normal control group and malignant glioma group, respectively. The Cos7 cells and the malignant glioma LN443 and U343 cells were cultured; the Cos7 cells were used as normal cell control group, and the LN443 and U343 cells as malignant glioma cell group. Western blotting method was used to detect the expression levels of SENP1, SENP2 and SENP6 proteins in human malignant glioma tissue and cells. Results: In brain tissue, the expression levels of SENP1,SENP2 and SENP6 proteins in malignant glioma group were higher than those in normal control group ( P<0. 05). Compared with normal cell control group, the expression levels of SENP1,SENP2 and SENP6 proteins in the LN443 and U343 cells in malignant glioma cell group were significantly increased (P<0. 05). Conclusion: SENP1, SENP2 and SENP6 proteins highly express in the malignant glioma tissue and cells,and they may play an important role in promoting the occurrence of malignant glioma.
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Objective:To observe the expressions of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells, and to elucidate the their effects in the development of malignant glioma.Methods:The samples of normal human brain tissue and malignant glioma tissue were obtained and used as normal control group and malignant glioma group, respectively.The Cos7cells and the malignant glioma LN443and U343cells were cultured;the Cos7cells were used as normal cell control group, and the LN443and U343cells as malignant glioma cell group.Western blotting method was used to detect the expression levels of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells.Results:In brain tissue, the expression levels of SENP1, SENP2and SENP6proteins in malignant glioma group were higher than those in normal control group (P＜0.05) .Compared with normal cell control group, the expression levels of SENP1, SENP2and SENP6proteins in the LN443and U343cells in malignant glioma cell group were significantly increased (P＜0.05) .Conclusion:SENP1, SENP2and SENP6proteins highly express in the malignant glioma tissue and cells, and they may play an important role in promoting the occurrence of malignant glioma.
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@#Background: There has been increasing evidence showing that stingless bee honey exhibits anti-oxidant, anti-inflammatory and anti-cancer properties. Pharmacologically-active components in honey such as flavonoids and phenolic constituents are known to contribute to its medicinal benefits. To the best of our knowledge, this is the first study on evaluating anticancer effects of locally-produced Malaysian stingless bee honey from Heterotrigona itama sp. on malignant glioma cells. Methods: Proliferation and apoptosis studies of U-87 MG cells following stingless bee honey treatment were carried out using MTS assay and acridine orange/propidium iodide dual staining, respectively. Results: Results demonstrated time and dose-dependent cytotoxicity using 0.625%, 1.25% and 10% stingless bee honey (P < 0.05). IC50 values were calculated using cells treated with 10% stingless bee honey. It was also observed that 10% stingless bee honey induced nuclear shrinkage, chromatin condensation and nucleus fragmentation, indicating that cellular changes were consistent with the apoptotic characteristics of the cells. Conclusion: These data provide a good basis for further evaluation of the medicinal properties of stingless bee honey from Heterotrigona itama sp. This source of honey may serve as a potential therapy for malignant glioma.
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@# Objective: To validate the effect and the possible mechanism of human regulatory factor X1 (RFX1) over-expression on the proliferation and invasion of glioma F98 cells. Methods: RFX1-overexpressed F98 cells (F98-RFX1 group) were constructed by lentivirus transfection, a control group (F98-Vector group) and normal group (F98 group) were established. The effect of RFX1 over-expression on F98 cell proliferation was observed with counting method, cell apoptosis was determined by AnnexinV-PI staining, and the cell invasion was observed with Transwell method. Results: F98 cell line over-expressing RFX1 was successfully established. The proliferation capacity of F98-RFX1 group was significantly lower than that of F98 group (48 h: [12.08±2.17]×104/ml vs [23.67±4.51]×104/ ml, P<0.05] and F98-Vector group (96 h: [8.17±0.31]×104/ml vs [18.58±1.18]×104/ml, P<0.05); The apoptosis level of cells in F98RFX1 group was significantly increased ([21.89±2.33]% vs [3.38±1.39]%, [10.42±1.83]%, P<0.05]; The invasiveness of cells in F98RFX1 group was significantly reduced ([33.3±7.99] vs [56.5±13.9], [60.6±11.8], P<0.01). Conclusion: RFX1 can regulate the expression of genes related with proliferation and invasion, thereby inhibiting the proliferation and invasion ability of glioma cells and promote cell apoptosis.
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Objective To investigate the autophagy and related mechanisms of human glioma U251 cells induced by temozolomide (TMZ) combined with radiotherapy.Methods MTT assay was used to detect the inhibitory effect of TMZ treatments with different concentrations (0～64 μ mol/L) on U251 cells for 24 h and 48 h respectively.The clone formation assay was used to detect the survival fraction of U251 cells under different concentrations of TMZ combined with radiotherapy,and the radiosensitization effect of TMZ was evaluated.The effect of the combined treatment on the apoptosis of U251 cells was detected by flow cytometry.The expression of microtubule-associated protein 1 light chain 3 (LC3)-Ⅰ,LC3-Ⅱ,Beclin-1 and phosphated protein kinase B (p-Akt) was detected by Western Blot.Results The inhibitory effect of TMZ on the proliferation of U251 cells was concentration-and time-dependent,and the IC50 values were 42.25 μmol/L and 25.13 μmol/L,respectively for the 24 h and 48 h treatment.TMZ has a radiosensitizing effect on U251 cells.TMZ combined with radiotherapy can induced apoptosis of U251 cells,and significantly up-regulate the expression levels of LC3-Ⅰ,LC3-Ⅰ and Beclin-1,and the differences were statistically significant (all P＜0.01).This combined treatment can down-regulate the expression of p-Akt protein,and the difference was statistically significant (P＜0.05).Conclusions TMZ combined with radiotherapy can activate autophagy on U251 cells.The mechanism may involve the inhibition of phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway by inhibiting Akt phosphorylation,the promotion of the expression of autophagy marker proteins (LC3-Ⅱ,LC3-Ⅰ and Beclin-1),and subsequent autophagy activation playing an anti-tumor effects.
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The incidence of leptomeningeal dissemination (LMD) of anaplastic glioma has been increasing. LMD can be observed at the time of initial presentation or the time of recurrence. As a result of both rarity and unusual presentation, a standard therapy has not yet been suggested. In contrast to leptomeningeal carcinomatosis for systemic solid cancers, a relatively prolonged survival is observed in some patients with LMD of anaplastic gliomas. Treatment modalities include whole craniospinal irradiation, intra-cerebrospinal fluid (CSF) chemotherapy, and systemic chemotherapy. In some cases, response to temozolomide (TMZ), with or without combined radiation has been reported. Here, we report two cases of LMD of an anaplastic glioma. In one case LMD presented at the time of diagnosis, and in the other at the time of recurrence after radiation. CSF cytology was positive in both cases, and persisted in spite of intrathecal methotrexate chemotherapy. Later, TMZ was prescribed for progressing brain parenchymal lesions, and both radiological and cytological responses were obtained after oral TMZ treatment.
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Humans , Brain , Cerebrospinal Fluid , Craniospinal Irradiation , Diagnosis , Drug Therapy , Glioma , Incidence , Meningeal Carcinomatosis , Methotrexate , RecurrenceABSTRACT
OBJECTIVE:To study the effect of Elemene emulsion combined with ganciclovir on invasive ability and TIMP-1 mRNA expression in C6 brain tumor stem cells. METHODS:Cells with 3 generations were isolated and cultured from rats'C6 ma-lignant glioma cell lines,and immunocytochemistry was adopted to detect the protein expressions of stem cell markers CD133,Nes-tin. 20 rats were randomly divided into blank control group(normal saline),ganciclovir group(intragastrically administrated,216 mg/kg),Elemene emulsion injection group (intravenously injected in tail,36 mg/kg) and combination group (intragastrically ad-ministrated 216 mg/kg of ganciclovir+intravenously injected 36 mg/kg of Elemene emulsion injection in tail),5 in each group, once every day,for 10 d. After 2 h of last administration,the blood of rats in each group was collected,serum was isolated and co-cultured 24 h with C6 BTSCs in in vitro culture medium. Boyden invasion test was used to detect the invasive ability of C6 BTSCs,and real-time quantitative polymerase chain reaction(RT-PCR)method was used to determine the TIMP-1 mRNA expres-sion of C6 BTSCs. RESULTS:Cells with 3 generations had obvious protein expressions in CD133 and Nestin,indicating they were BTSCs. Compared with blank control group,the cell invasion rates of C6 BTSCs in ganciclovir group,Elemene emulsion in-jection group and combination group were obviously decreased,TIMP-1 mRNA expression was obviously enhanced,with statisti-cal significances(P<0.05). Compared with ganciclovir group and Elemene emulsion injection group,the cell invasion rate of C6 BTSCs in combination group was obviously decreased,TIMP-1 mRNA expression was obviously enhanced,with statistical signifi-cances(P<0.05). CONCLUSIONS:Elemene emulsion injection combined with ganciclovir can obviously inhibit the cell invasion of C6 BTSCs,the mechanism may be associated with promoting the TIMP-1 generation,and combination use shows better effect than single use.
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Malignant glioma cells invading surrounding normal brain are inoperable and resistant to radio- and chemotherapy, and eventually lead to tumor regrowth. Identification of genes related to motility is important for understanding the molecular biological behavior of invasive gliomas. According to our previous studies, Metallothionein 1E (MT1E) was identified to enhance migration of human malignant glioma cells. The purpose of this study was to confirm that MT1E could modulate glioma invasion in vivo. Firstly we established 2 cell lines; MTS23, overexpressed by MT1E complementary DNA construct and pV12 as control. The expression of matrix metalloproteinases (MMP)-2, -9 and a disintegrin and metalloproteinase 17 were increased in MTS23 compared with pV12. Furthermore it was confirmed that MT1E could modulate MMPs secretion and translocation of NFkB p50 and B-cell lymphoma-3 through small interfering ribonucleic acid knocked U87MG cells. Then MTS23 and pV12 were injected into intracranial region of 5 week old male nude mouse. After 4 weeks, for brain tissues of these two groups, histological analysis, and immunohistochemical stain of MMP-2, 9 and Nestin were performed. As results, the group injected with MTS23 showed irregular margin and tumor cells infiltrating the surrounding normal brain, while that of pV12 (control) had round and clear margin. And regrowth of tumor cells in MTS23 group was observed in another site apart from tumor cell inoculation. MT1E could enhance tumor proliferation and invasion of malignant glioma through regulation of activation and expression of MMPs.
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Animals , Humans , Male , Mice , B-Lymphocytes , Brain Neoplasms , Brain , Cell Line , DNA, Complementary , Drug Therapy , Glioma , Matrix Metalloproteinases , Metallothionein , Mice, Nude , Nestin , RNAABSTRACT
OBJECTIVE:To observe clinical effect and safety of temozolomide (TMZ) combined with radiotherapy in the treatment of malignant glioma. METHODS:58 patients with malignant glioma were randomly divided into TMZ-RT group(32 cas-es)and RT group(26 cases). RT group were given the therapy of 3D-CRT 1.8-3 Gy/d,and some patients received the hyperfrac-tion radiotherapy with total dose of 60-75 Gy. TMZ-RT group were given TMZ chemotherapy 75 mg/(m2·d)combined with radio-therapy for 6 weeks and TMZ sequential therapy 150-200 mg/(m2·d)for 4 weeks. Therapeutic efficacy and ADR were observed in 2 groups. RESULTS:After treatment,in TMZ-RT group,9 cases had complete remission,11 cases had partial remission,10 cas-es were stable and 2 cases were progressive;in RT group,6 cases had complete remission,8 cases had partial remission,8 cases were stable and 4 cases were progressive. There was no statistical significance in short-term efficacy between 2 groups(P>0.05). The 1,2,3-year survival rates of TMZ-RT group were 81.3%(26/32),78.1%(25/32) and 62.5%(20/32),while those of RT group were 84.6%(22/26),34.6%(9/26)and 15.4%(4/26). There were statistical significance in 2,3-year survival rates between 2 groups (P<0.05). Median recurrent time of TMZ-RT group was (23.2 ± 8.6) months,while that of RT group was (15.6 ± 8.7) months,with statistical significance(P<0.05). According to the KPS,TMZ-RT group was(89.6±9.6)and RT group(65.1±10.1), with statistical significance (P<0.01). The side effect of 2 groups were slight. CONCLUSIONS:Radiotherapy combined with TMZ can significantly improve the survival rates of 2 and 3-year and delay the tumor recurrent time,and lighter adverse reaction. It can be used as new therapy method for malignant glioma. But it needs further study for treating the malignant glioma patients with osteomyelitis or low immune function.
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Objective To compare with the therapy effect of bevacizumab or bevacizumab combined with dose-dense temozolomide on recurrent malignant gliomas.Methods Thirty-eight patients with recurrent malignant glioma,admitted to our hospital from June 2008 to June 2012,were chosen in our study.Among them,26 patients accepted bevacizumab therapy (single drug group) and 12 accepted bevacizumab combined with dose-dense temozolomide therapy (unite drugs group).Then,the clinical data,progressive free survival (PFS) and adverse effects were retrospectively analyzed.Results Single drug group:the median PFS was 21.8 weeks,PFS-6 months was 37.8％;united drug group:the median progressive free survival was 34.7 weeks,PFS-6 months was 52.7％;significant differences were noted between the two groups (x2=8.161,P=0.023) Conclusion Bevacizumab can prolong the PFS of patients with recurrent malignant glioma;the therapy effect of bevacizumab and dose-dense temozolomide is better than single drug,which can be recommended as an effective therapy for recurrent malignant glioma.
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Objective To observe the clinical effect of temozolomide combined with radiotherapy in the treatment of malignant gliomas. Methods A total of 64 patients with pathologically confirmed glioblastoma received radiotherapy combined with chemotherapy in our hospital from January 2008 to January 2014, radiotherapy synchronized with TMZ chemotherapy were used in 39 patients (synchronized group) and radiotherapy sequential with TMZ chemotherapy in 25 patients (sequential group).1 year and 3 year survival rates, response rate and drug safety were compared between the two groups. Results The patients were followed up for 6 months to 3 years, the synchronous group of patients 1 years and 3 years survival rates (74.4%, 38.5%) were higher than the sequential group(64.0%, 28.0%) (P<0.05), objective re-sponse rate of two groups were 64.1%and 44.0%, the difference was statistically significant(P<0.05), gastrointestinal re-action rates of two groups were 23.1% and 20.0%, the difference was not statistically significant (P>0.05). Adverse re-actions were statistically significant (P<0.05). Conclusion The effect of radiotherapy synchronized combined with TMZ chemotherapy is better than radiotherapy sequential with TMZ chemotherapy for glioblastoma, it is still the first choice for the treatment of glioblastoma although the clinical manifestations of toxicity increased than the sequential group.
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PURPOSE: The aim of this study was to assess the feasibility and efficacy of hypofractionated simultaneous integrated boost-intensity modulated radiotherapy (SIB-IMRT) using three-layered planning target volumes (PTV) for malignant gliomas. MATERIALS AND METHODS: We conducted a retrospective analysis of 12 patients (WHO grade IV-10; III-2) postoperatively treated with SIB-IMRT with concurrent temozolomide. Three-layered PTVs were contoured based on gadolinium-enhanced magnetic resonance imaging as follows; high risk PTV (H-PTV) as the area of surgical bed including residual gross tumor with a 0.5 cm margin; low risk PTV (L-PTV) as the area surrounding the high risk PTV with 1.5 cm margin; moderate risk PTV (M-PTV) as a line at one-third the distance from high risk PTV to low risk PTV. Total dose to high risk PTV was 70 Gy in 8 and 62.5 Gy in 4 patients. RESULTS: The median follow-up time was 52 months in surviving patients. The 2- and 5-year overall survival (OS) rates were 66.6% and 47.6%, respectively. The 2- and 5-year progression-free survival (PFS) rates were 57.1% and 45.7%, respectively. The median OS and PFS were 48 and 31 months, respectively. Six patients (50%) progressed: in-field only in one, out-field or disseminated in 4, and both in one patient. All patients completed planned treatments without a toxicity-related gap. Asymptomatic radiation necrosis was observed in 4 patients at post-radiotherapy 9-31 months. CONCLUSION: An escalated dose of hypofractionated SIB-IMRT using three-layered PTVs can be safely performed in patients with malignant glioma, and might contribute to better tumor control and survival.
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Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Glioma/radiotherapy , Radiotherapy, Intensity-Modulated/methods , Retrospective StudiesABSTRACT
Objective To investigate the coping mode and quality of life of patients with malignant glioma during chemotherapy.Methods European organization for research and treatment of cancer quality of life,Core 30(EORTC QLQ-C30) and medical coping modes questionnaire(MC-MQ)were used for the survey among 45 patients with malignant glioma during chemotherapy.Results The scores on patients' emotion function and role function were(7.80±2.52)and(16.26±5.35),both at a low level.The avoidance factor in copy mode was scored(16.26±5.35),significantly lower than the norm(14.44±2.97)(P<0.05). Conclusion Nurses should make pertinent measures for intervention,encouraging patients to take right cope mode and keeping positive attitude so as to improve their quality of life.
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Objective To observe the efficacy and complications of iodine-125 radioactive seeds implantation after surgery of recurrent malignant gliomas.Methods Fifty-eight patients with recurrent gliomas,admitted to our hospital from September 2006 to January 2010,were randomly divided into observation group (n=26) and control group (n=32).All patients accepted tumor subtotal resection.Patients in the observation group were implanted with iodine-125 seeds which were planned by therapy planning system (TPS) depending on the tumor size and position.Patients in the control group were irradiated at 50-60 Gy after the operations.Patients from both groups were followed up for 3 years,and the 1-year and 3-year survival rates,median survival time,complications and quality of life were observed.Results The survival rate of 1-year of observation group and control group was 46.2％ and 34.4％,without significant difference (P＞0.05),while that of 3-years time was 19.2％ and 12.5％,with significant difference (P＜0.05).The average survival time of the two groups was (15.6±2.5) months to (9.4±1.3) months,with statistically significant difference (P＜0.05).No severe complications (grade four or higher than grade four) were noted; lower rates of dizziness and headache were found in observation group as compared with those in control group (P=0.030,P=0.035).Conclusions The iodine-125 radioactive seeds implantation after surgery of recurrent malignant gliomas not only can improve the survival rate and prolong survival time,but also reduce radiation brain damage to improve the life quality.So it is a safe and effective treatment,and worth of applicattion in clinic.
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Introducción: la asociación de un cáncer con un embarazo pone en juego dos pronósticos vitales: el de la madre y el del feto. Objetivo: describir un caso clínico de una paciente con tumor cerebral asociado al embarazo. Métodos: paciente femenina de 34 años de edad, nulípara, que a las 25 sem de embarazo se le diagnostica un tumor cerebral y se le realiza craniectomía de fosa posterior, bulbotomía y evacuación del quiste en el hospital "Hermanos Ameijeiras", en La Habana. A las 31 sem de edad gestacional fue remitida al hospital "Ramón González Coro" para vigilancia del bienestar fetal y materno con la recomendación de cesárea electiva si es posible al término de la gestación por indicación neurológica. Se utilizaron los inductores de la madurez pulmonar fetal por la posibilidad de terminar la gestación antes del término. Progresa sin incidentes hasta las 38 sem de gestación, momento en que se le practica la cesárea. Resultados: se obtuvo un recién nacido vivo, femenino, con peso de 2 600 g y apgar 9/9. La madre y el neonato evolucionaron satisfactoriamente hasta el alta. Se remitió a la madre nuevamente al hospital "Hermanos Ameijeiras" para su control y seguimiento posoperatorio. Conclusiones: la presencia de una tumoración cerebral asociada al embarazo implica un alto riesgo de morbilidad y mortalidad materna y perinatal, por lo que requiere de un manejo multidisciplinario para lograr óptimos resultados.
Introduction: the association of cancer with a pregnancy involves two vital forecasts: mother and her fetus. Objective: to describe a case of a patient with brain tumor associated with pregnancy. Methods: A 34 year- old female patient, who was nulliparous, and at 25 weeks of pregnancy was diagnosed with a brain tumor. She underwent a posterior fossa craniectomy, bulbotomy and cyst evacuation at Hermanos Ameijeiras hospital in Havana. At 31 weeks of her gestational stage, she was sent to the Ramón González Coro Maternity hospital for monitoring fetal and maternal well-being with the recommendation of elective cesarean if possible at the end of gestation as it was neurologically indicated. Lung maturity fetal inducers were used by the possibility of terminating this pregnancy before term. This pregnancy uneventfully progressed until 38 weeks of gestation, when Caesarean section was conducted. Results: a live female infant was obtained; she weighed 2 600 g and had Apgar 9/9. Mother and infant recovered satisfactorily to discharge. This mother was referred back to Hermanos Ameijeiras hospital for postoperative monitoring and control. Conclusions: the presence of a brain tumor associated with pregnancy carries a high risk of morbidity and maternal and perinatal mortality, therefore this issue requires a multidisciplinary approach to achieve optimal results.
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Objective:The poor prognosis of patients with malignant gliomas (MG) has led to the search for new therapeutic strat-egies. Recently, nimotuzumab has been studied as a new anti-EGFR-receptor humanized monoclonal antibody in patients with MG, who showed improvement of outcome and good tolerability. We conducted phase I of our study to determine the toxicity, tolerated dose, and clinical feasibility of nimotuzumab in combination with concurrent chemoradiotherapy for Chinese MG patients after surgical resection. Methods:Patients with pathologically proven grades 3 and 4 glioma were enrolled in the study. The protocol included infu-sions of nimotuzumab plus standard Stupp schedule (postoperative radiotherapy in a total dose of 60 Gy in combination with daily te-mozolomide). Patients received 6 weekly infusions of nimotuzumab at three levels (100, 200, and 400 mg/week). If none of the first three patients enrolled at a dose level experienced dose-limiting toxicity (DLT), the dose was increased, as appropriate. If DLT was ob-served, another three patients were added to the dose level. Results:Nine patients with MG were enrolled, including 7 with grade 3 MG and 2 with glioblastoma. The treatment was well tolerated, and no evidence of grade 3 or 4 adverse events was detected, even at the highest level (400 mg/week). Grade 1 or 2 myelosuppression was the most common toxicity. Three months after treatment, stable dis-ease occurred in 5 patients, whereas progression disease was observed in 4 patients. Conclusion:Nimotuzumab combined with concur-rent chemoradiotherapy was associated with mild toxicity in Chinese MG patients.
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Objective To investigate the effects of enterovirus 71 (EV71) on growth,apoptosis and human telomerase reverse transcriptase (hTERT) expression of human malignant glioma cell line U251 in vitro.Methods U251 cells were cultured in RPMI 1640 for 24 hours,then randomly divided into experimental group and control group.In the experimental group,EV71 were added in cell culture holes at multiplicity of infection (MOI) equal to 1,and cultured continuously for 72 hours with U251 cells,but in the control group,EV71 were not put.The morphological features and growth of cells were observed under inverted microscope at 0 hour,24 hours,48 hours and 72 hours of EV71 treatment,respectively.At the same time,cell apoptosis was measured by flow cytometry using Annexin V/PI double staining method.hTERT expression was detected with semi-quantitative reverse transcriptase-polymerase chain reaction (RTPCR).Results The growth of U251 cells was inhibited significantly at 24 h of EV71 treatment under inverted microscope in experimental group when compared with control group,and at 48 h and 72 h,the morphological features of these cells became significantly shrunken,smaller and irregular shape in experimental group,but which were uniform size in control group.At 24 h,48 h,72 h of EV71 treatment,flow cytometry showed the apoptosis rates were significantly higher in experimental group than that in control group [24 h:(12.55 ± 2.38) ％ vs (1.42 ± 0.21) ％,48 h:(65.60 ± 8.48)％ vs (1.42 ±0.17)％,72 h:(87.52 ±3.05)％ vs (1.41 ±0.16)％,all P =0.000],and there was upward trend day by day,but no change in control group.At the same time,hTERT mRNA expression levels of U251 cells were significantly lower in experimental group than control group [24 h:(0.58 ± 0.05) vs (0.89 ± 0.05),48 h:(0.23 ± 0.04) vs (0.89 ± 0.03),72 h:(0.10 ± 0.03) vs (0.90 ± 0.06),all P =0.000],and the downward trend was observed day by day,but there was no this trend in control group.Conclusions EV71 can effectively inhibit the growth of U251 cells,induce the cell apoptosis,and has the potential anti-tumor effect.Its mechanisms may be partly related to down regulation the hTERT expression of U251 cells.
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Objective To show evidences that autophagy is induced in human malignant glioma cell line U251 by enterovirus(EV71) and its effect on the expression of microtubule-associated protein 1 lightchain 3 (LC3) in vitro.Methods U251 cells were cultured in RPMI 1640 for 24 hours,then randomly divided into experimental group and control group.In experimental group,EV71 were added in cell culture holes at multiplicity of infection (MOI) equal to 1,and cultured continuously.After 12 hours post infection of U251 cells with EV71,autophagic vacuoles of U251 cells were marked by monodansylcadaverine staining and observed under fluorescence microscope.After 24 hours post infection,expression and intracellular distribution of LC3 protein in U251 cells were observed under fluorescence microscope by immunofluorescence.Expressions of LC3-I and LC3-Ⅱ protein were measured by Western blot and analysis of LC3-Ⅱ protein expression was performed with semi-quantitative calculation at 2,4,8,12,24 and 48 hours post infection of U251 cells with EV71,respectively.Results Autophagic vacuoles stained by MDC in U251 cells appeared as distinct dot-like structures distributed under fluorescence microscope.The number of autophagic vacuoles were increased significantly at 12 hours post infection of U251 cells with EV71 when compared with control group.The morphological features of these cells became significantly shrunken,smaller and irregular shape at 24 hours post infection of U251 cells,and the expressions of LC3 protein were significantly higher in experimental group than those in control group.Under a fluorescence microscope,LC3 protein distributed within the cytoplasm or localizing in the perinuclear regions.At 4 hours post infection of U251 cells with EV71,the expression of LC3-Ⅱ protein started to increase,and was significantly higher than that in control group (P ＜ 0.01).Conclusion These results indicate that EV71 can effectively induce autophagy of human malignant glioma cell line U251,and play its oncolytic effect.