ABSTRACT
@#[Abstract] Objective: To investigate the miR-423-5p expression in brain glioma tissues and cell lines, and its promotive effect on temozolomide (TMZ) chemoresistance by targeting PDCD5 (programmed cell death protein 5). Methods: Tumor tissues and matched peritumoral tissues were collected from 20 brain glioma patients who were surgically treated in the Department of Neurosurgery, Affiliated Hospital of Beihua University between January 2017 and December 2018. Glioblastoma cell lines (U251, U87, SHG-44) and human normal glial cell line HMC-3 were also used in the study. The relative expression of miR-423-5p and PDCD5 in brain glioma and peritumoral tissues and cell lines was detected by qPCR. The synthesized miR-423-5p mimics and miR-NC were respectively transfected into U251 and U87 cells; meanwhile, TMZ at different concentrations (50, 100, 150 and 200 μmol/L) were also used to treat the cells. Then, the chemoresistance of cells to TMZ were determined. MTT assay and colony formation assay were used to examine the proliferation of U251 and U87 cells, andWestern blotting was used to detect the expression of c-caspase 3, Bcl-2 and PDCD5 proteins in U251 and U87 cells. The targeting relationship between PDCD5 and miR-423-5p was validated through Dual luciferase reporter gene assay. Results: miR-423-5p was highly expressed in glioma tissues and glioma cell lines (all P<0.01). As compared with the miR-NC group, the proliferation and TMZ-chemoresistance of U251 and U87 cells in miR-423-5p mimics group significantly increased (all P<0.01). Dual luciferase reporter gene assay validated that miR-423-5p could bind with PDCD5 3' UTR to suppress the expression of PDCD5. Conclusion: High expression of miR-423-5p enhances the chemoresistance of glioma cells to TMZ, and miR-423-5p may serve as a potential therapeutic target in the treatment of brain glioma.
ABSTRACT
Introduction@#Wilms’ tumor 1gene was originally discovered as mutated in nephroblastoma, a common pediatric kidney cancer also known as Wilms’ tumor. This gene’s product alteration was indicating the safety of WT1 immunotherapy as well as a potential therapeutic response to its application in patients with glioma. @*Goal@#Our aim was to further elucidate the role of WT1 as a diagnostic and prognostic marker in brain glioma in neuropathology field.@*Materials and Methods@#In this study, formalin fixed paraffin embedded blocks of 135 patients with brain glioma were selected. After tissue preparation for WT1 immunohistochemical evaluation, 2 tissue preparations were excluded due to unsatisfactory amount of tissue. Therefore, data about tissue specimens from 133 patients were included in statistical analysis.@*Results@#In this study, out of 133 cases, 55 were astrocytomas, 42 were oligodendroglioma, 35 were glioblastoma and 1 was mixed oligoastrocytoma.WT1 immunohistochemistry expression was found in 127/6 (95.5%) samples. For the glioblastoma, WT1-expression significantly increased with patient’s age (p=0.05, table 3). WT1 expression and Ki67 proliferation index had prognostic effect in patients with brain glioma (p<0.05), and low expression mean survival was 48.5 months, high expression survival was 18.4 months respectively.@*Conclusion@#WT1 expression in Mongolian patients with brain glioma had significantly associated with several adverse prognostic indicators including high Ki67 proliferation index (high grade tumor) and high expression of WT1 and univariable survival.
ABSTRACT
Focus on the inconsistency of the shape, location and size of brain glioma, a dual-channel 3-dimensional (3D) densely connected network is proposed to automatically segment brain glioma tumor on magnetic resonance images. Our method is based on a 3D convolutional neural network frame, and two convolution kernel sizes are adopted in each channel to extract multi-scale features in different scales of receptive fields. Then we construct two densely connected blocks in each pathway for feature learning and transmission. Finally, the concatenation of two pathway features was sent to classification layer to classify central region voxels to segment brain tumor automatically. We train and test our model on open brain tumor segmentation challenge dataset, and we also compared our results with other models. Experimental results show that our algorithm can segment different tumor lesions more accurately. It has important application value in the clinical diagnosis and treatment of brain tumor diseases.
Subject(s)
Humans , Brain Neoplasms , Diagnostic Imaging , Glioma , Diagnostic Imaging , Image Processing, Computer-Assisted , Magnetic Resonance Imaging , Neural Networks, ComputerABSTRACT
Objective To investigate the anti-glioma treatment effect of triptolide modified by phospholipid complex nanocapsule encapsulation technology in vitro.Methods Human glioma U87 ceils were used to investigate the uptake efficiency of tumor cells to nano-microcapsules TPL and TPL.MTT method was used to detect the proliferation inhibition rate of U87 cells by nano-microcapsule TPL.The levels of ATP and ROS in U87 cells were measured to reflect cell proliferation and apoptosis effect of nano-microcapsules TPL.Results Glioma cells are sensitive to TPL and nano-microcapsule TPL.Compared with single TPL,nano-microcapsule TPL has better glioma cell inhibitory effect and anti-tumor activity (P＜0.05),and has significantly reduced ATP level (P＜0.05) and increased ROS level (P＜0.05) in U87 cells.Conclusions After being encapsulated by nanocapsule technology,TPL has good glioma cell targeting and can significantly enhance the inhibitory effect of drug-loaded liposomes on glioma cell proliferation.
ABSTRACT
Objective To evaluate the targeting and inhibitory effects of the targeted artemether liposomes modified with APGA- PEG2000-DSPE conjugate in vitro. Methods APGA was used as targeted molecular, and the new functional material APGA- PEG2000-DSPE conjugate was synthesized and confirmed by using MALDI-TOF-MS. A kind of targeted artemether liposomes was developed by modifying APGA-PEG2000-DSPE using film dispersed method. Co-culture model of BBB-glioma C6 tumor spheroids was developed, and it was used to study the transporting efficiency across the BBB and the penetration ability of the liposomes into glioma C6 tumor spheroids viewed by a confocal laser scanning microscope. Flow cytometry was used to evaluate the targeting properties of the preparations to C6 glioma cells. SRB method was used to study the inhibitory effect of targeted liposomes on the growth of C6 glioma cells. Results The APGA-PEG2000-DSPE conjugate was confirmed by using MALDI-TOF-MS, and its average mass was 3 150. The co-culture model showed that the targeted liposomes can better transport across the BBB and then penetrate into the glioma C6 tumor spheroids. To C6 glioma cells, the IC50 of the targeted artemether liposomes, artemether liposomes and free artemether were 45, 82, and 95 μmol/L, separately. Conclusion The targeted artemether liposomes could effectively transport across the BBB and penetrate into the glioma C6 tumor spheroids. The targeted artemether liposomes had stronger inhibitory effect on brain glioma cells in vitro.
ABSTRACT
Objective:To investigate the clinical efficacy of stereotactic radiation therapy combined with temozolomide on recurrent glioma.Methods:A total of 36 patients with recurrent glioma were retrospectively analyzed and divided into a control group (n=12),who received stereotactic radiation therapy,and an experimental group (n=24),who received stereotactic radiation therapy plus temozolomide.The clinical efficacy and adverse reactions for the 2 groups were compared.Results:Total effective rate and local control rate for clinical treatment were 66.67％ and 93.94％,respectively.Late adverse reaction was not observed.The effective rate and local control rate in the experimental group were 77.27％ and 95.45％,which were slight higher than those in the control group,with no statistical significance (P＞0.05).The 0.5-,1-,2-,3-year follow-up total survival rates were 90.91％,63.64％,42.42％,and 15.15％,respectively.The 0.5-,1-,2-,3-year follow-up survival rates in the experimental group were 95.45％,72.72％,54.54％ and 22.73％,respectively,while those in the control group were 81.82％,45.45％,18.18％,and 0％,respectively.Survival analysis showed the survival time for the experimental group was significantly longer than that of the control group (30.00 months vs 14.00 months,P=0.010).Conclusion:Stereotactic radiation therapy combined with temozolomide for recurrent glioma is effective,and it has positive effect on improving the clinical efficacy and survival rate for the patients.
ABSTRACT
Objective Cell cycle-associated protein 1 (Caprin-1) is closely related to the development and progression of cancer. This study aimed to explore the expression of Caprin-1 in the clinical glioma specimen and its influence on the biological char-acteristics of the glioma cell line. Methods Brain tissue specimens were collected from 29 glioma patients and 2 normal humans that died of accidental trauma. A stably transfected U251 cell line with overex-pressed Caprin-1 was established,and the U251 cells were transfected with the pEGFP-C1 plasmid (the negative control group), or the pEGFP-C1-Caprin-1 plasmid (the experimental group), or left un-transfected (the blank control group). The expressions of Caprin-1 mRNA and protein in the cells were determined by RT-PCR and Western blot, and the proliferation and migration of the cells exam-ined by scratch test and Transwell assay,respectively. Results The expression of Caprin-1 was upregulated with the increased grade of glioma,145.9±22.0,444.4±110.0,and 1661.0±54.5 in WHO gradeⅡ,Ⅲ,andⅣglioma,respectively,significantly higher than in the normal brain tissue (P<0.05). Both the mRNA and protein expressions of Caprin-1 were remarkably higher in the experimental group (1.70±0.19 and 1.07±0.09) than in the blank control(0.89±0.10 and 0.52±0.04) and negative control(0.98±0.08 and 0.58± 0.03) (P<0.05).The A value was also markedly higher in the former group(2.55±0.14) than in the latter two(1.40±0.06 and 1.35± 0.04) (P<0.01),and so were the count of migrated cells(526.00±42.19 vs 289.00±29.24 and 279.00±32.48,P<0.01) and the ex-pression of CyclinD1 (0.60±0.05 vs 0.13±0.03 and 0.15±0.05, P<0.01). Conclusion The expression of Caprin-1 in the U251 cells was upregulated with the increased WHO grade of glioma,and the overexpression of Caprin-1 accelerated the proliferation and mi-gration of the U251 cells.
ABSTRACT
18F-FET PET imaging had its fundamental principle introduced, and then had its application to brain glioma diagnosis described from the aspects of patient survival period estimation, facilitating radiotherapy planning, post-radiotherapy effect monitoring and pseudoprogression neuroimaging evaluation. The advantages and disadvantages of 18F-FET PET were analyzed when used to diagnose brain glioma, and it's pointed out that 18F-FET PET combined with structural imaging technologies such as MRI spectral analysis, perfusion and diffusion-weighted imaging contributed to enhancing the accuracy of brain glioma diagnosis, positioning, differentiating, grading, prognosis prediction and treatment.
ABSTRACT
Objective To investigate the effects of VCAM-1 on migration and invasion of glioma cell lines . Methods The techniques of lentivirus pSGU6/GFP/Neo-based VCAM-1 shRNA and EF1 a-GFP/puro-based VCAM-1 expression vector, the scratch wound healing migration and transwell invasion assays , and the Western blot and cell staining were applied to observe the effects of VCAM-1 expression levels on migration and invasion of glioma cell line cells.There are four groups in T98G cells including control, vector, scramble and shRNA-VCAM-1 groups and three groups in U251 cells covering control, vector and VCAM-1 overexpressed groups ( n=6 per group) .Results The stabled glioma cell lines of T98 G cells with down-regulated VCAM-1 and U251 cells with VCAM-1 overexpression were established by using lentivirus-based VCAM-1 shRNA and expression vector.The ability of scratch wound healing (migration activity) decreased significantly (P<0.01) in T98G cells with lower VCAM-1 expression levels, while the migration activity was obviously improved in U251 cells with overexpressed VCAM-1 ( P <0.05 ) .Similarly, the invasion ability was significantly inhibited ( P <0.05) in T98G cells with silenced VCAM-1, as well as VCAM-1 overexpression could enhance the invasion ability of U251 cells ( P<0.01 ) .Conclusions VCAM-1 improves the migration activity and invasion ability of human glioma cell line cells.
ABSTRACT
Objective To investigate the correlation between the levels of serum tumor necrosis factor-α(TNF-α),glial fibrillary acidic protein(GFAP) and the malignant degree and prognosis of patients with brain glioma.Methods A total of 90 patients with brain glioma were collected from March 2013 to September 2015 in Shaanxi Provincial People's Hospital.The patients were divided into low malignant degree group (n =43) and high malignant degree group (n =47) according to the pathological grade of malignant degree.Fifty-two healthy volunteers were selected as control group at the same period.The fasting venous blood 5 mL of the patients with brain glioma was collected at the time points of pretherapy,the 14th day after operation,the first day after radiotherapy and chemotherapy.The fasting venous blood 5 mL was collected on the day of physical examination in the control group.The level of serum TNF-α was detected by enzyme-linked immunosorbent assay,and the level of serum GFAP was measured by double antibody sandwich method.Results The levels of serum TNF-α and GFAP in the low malignant degree group and high malignant degree group were significantly higher than those in the control group (P ＜ 0.05),and the levels of serum TNF-α and GFAP in the high malignant degree group were significantly lower than those in the low malignant degree group before treatment (P ＜ 0.05).The levels of serum TNF-α and GFAP at the time points on the 14th day after operation and after radiotherapy and chemotherapy were significantly lower than those before treatment (P ＜ 0.05);the levels of serum TNF-α and GFAP after radiotherapy and chemotherapy were significantly lower than those on the 14th day after operation in the low malignant degree group (P ＜ 0.05).The level of serum TNF-α on the 14th day after operation was significantly lower than that before treatment in the high malignant degree group (P ＜ 0.05).There was no significant difference in serum GFAP level between the 14th day after operation and before treatment in the high malignant degree group (P ＞ 0.05).The levels of serum TNF-α and GFAP after radiotherapy and chemotherapy were significantly lower than those before treatment and the 14th day after operation in the high malignant degree group (P ＜ 0.05).The levels of serum TNF-α and GFAP in the patients with complete remission were significantly lower than those in the patients with partial remission,stable disease and progressive disease after treatment (P ＜ 0.05).The levels of serum TNF-α and GFAP in the patients with partial remission were significantly lower than those in the patients with stable disease and progressive disease after treatment (P ＜ 0.05).The levels of serum TNF-α and GFAP in the patients with stable disease were significantly lower than those in the patients with progressive disease after treatment (P ＜ 0.05).Conclusions The levels of serum TNF-α and GFAP are correlated with the malignant degree and the therapeutic effect in patients with brain glioma.It can be used as a biological marker to evaluate the malignant degree,therapeutic effect and prognosis of brain glioma.
ABSTRACT
To investigate the effect of MMP-26 on human glioma angiogenesis and the possible mechanism.Methods The MMP-26 plasmid and empty plasmid pcDNA3.1 were stably transfected into U251 cells to establish a nude mice xenograft model,and then an in vitro human tumor tissue-based three-dimensional angiagenic model.Tissue disks were visually assessed over time to determine the percentage of wells that developed an angiogenic response(I％) and the density and length of neovessel growth were graded at intervals using a semiquantitative visual growth-rating scheme (angiogenic index,AI,0-16scale) in groups of MMP-26 transfected U251 cells (U251-MMP-26),pcDNA3.1 vector-transfected U251 cells (U251-pcDNA3.1) and non-transfected U251 cells (U251).RT-PCR and immunohistochemistry were used to detect the expression of mRNA and protein of MMP-26 and VEGF in groups of U251-MMP-26,U251-pcDNA3.1 and U251.Immunohistochemical localization of CD31 was determined in the endothelial tubes invading the fibrin-thrombin clot matrix.Results Immunohistochemical endothelial cell markers CD31 was positive in the vascular tubes invading the fibrin-thrombin clot matrix,confirming their endothelial origin.The angiogenesis results showed that difference of length of micro capillaries,density of branches,and the area occupied between U251-MMP-26 groups and control groups were significant.The percentage of tumor implants that developed invasion (I％) and the angiogenic index AI in U251-MMP-26 group on day 14 were higher than those of U251-pcDNA3.1 group and U251 group (P ＜ 0.05).The trends of I％ and AI in 14 days were significant compared with those in control groups.The expression of mRNA and protein of MMP-26and VEGF in U251-MMP-26 group was significantly higher in U251-MMP-26 group than those in U251-pcDNA3.1 group and U251 group(P ＜0.01).Conclusion The effect of MMP-26 on promoting glioma angiogenesis may be related to the increased expression of VEGF,which can be used as targets for anti-tumor therapy.
ABSTRACT
Objective To evaluate the relationship between 1H-MRS features of brain glioma and its pathological grade and invasiveness, and the correlation between those features and the expression of VEGF, MMP-9 and uPA.Methods The brain gliomas in 55 patients confirmed by pathology were divided into two groups: high grade and low one.An analysis of intragroup and intergroup differences in some metabolite ratios of 1H-MRS in the tumor parenchyma was conducted.The expression of MMP-9,VEGF and uPA in brain glioma was detected,and the correlation between the above three parameters and changes in metabolite ratios of 1H-MRS were explored.Results There was significant difference and correlation in Cho/Cr and Cho/NAA ratio in the tumor parenchyma between high and low grade(P0.05).Conclusion VEGF and MMP-9 play an important role in the development of brain glioma.Cho/Cr and Cho/NAA ratios provide an important reference for preoperative grading of brain gliomas and indirectly reflect the expression of VEGF and MMP-9.
ABSTRACT
Objective To synthesize a new kind of acid-sensitive doxorubicin prodrug nanoparticles and to evaluate its anti-brain glioma effect and efficiency through blood-brain barrier (BBB). Methods The prodrug acid-sensitive poly-ethylene glycol (PEG)-doxorubicin (PEG-DOX) copolymer was synthesized by Schiff base reaction, and PEG-DOX pro-drug nanoparticles (PEG-DOX NPs) were prepared by self-assembling. The character of PEG-DOX copolymer was detected by dynamic light scattering (DLS) instrument and 1H NMR. The morphology of PEG-DOX NPs was observed by transmission electron microscopy (TEM). The character of drug release was detected by UV mothed. The cellular uptake efficiency of glio-ma cells to PEG-DOX NPs was observed by inverted fluorescence microscope. The anti-brain glioma effects of PEG-DOX NPs and Free DOX were studied by MTT mothed. PS80-PEG-DOX NPs were gained by the modification of PEG-DOX NPs with Tween 80. Nine BALB/c mice were separated into Free DOX, PEG-DOX NPs and PS80-PEG-DOX NPs groups by ran-dom drawing lots. The mean fluorescence intensity of brain and main organs were observed by in vivo imaging system. Re-sults The copolymer of PEG-DOX can self-assemble into nanoparticles with the diameter of 100 nm. PEG-DOX NPs can quickly release DOX in acid environment. Although PEG-DOX NPs had slow cancer cell uptake than Free DOX, it had lon-ger accumulation. MTT results showed that PEG-DOX NPs had concentration dependent anti-brain glioma effect. Indepen-dent samples t-test indicated that the efficiency through BBB was significantly higher in PS80-PEG-DOX NPs group than that of Free DOX group and PEG-DOX NPs group. Conclusion PEG-DOX NPs show well anti-brain glioma effect in vi-tro, and can across BBB with high efficiency after modification, which make it possible for a potential therapeutic prodrug for brain glioma.
ABSTRACT
Aim To establish ICR animal model with C6 glioma stem cells, to provide the ideal model for the further study of gli-oma stem cells in brain glioma model. Methods C6 glioma stem cell was cultured in vitro by suspension,and was identified with Nestin antibody. C6 stem cells of ICR mouse glioma model were used to investigate survival state and tumor volume in mice after the operation. HE staining and CD133 immunohistochemi-cal study were adopted to investigate the postoperative pathologi-cal changes in mice. Results The expression of Nestin was 96. 01% in C6 glioma stem cells, and Nestin was highly ex-pressed in the cultured C6 glioma stem cells. Mice were inocula-ted with tumor after loss of appetite, weight, behavior and slow, sluggish reaction. Tumor volume at day 21 after modeling was (9. 77 ± 6. 58) mm3 . After HE staining, the model showed the invasive growth, tumor cell shrinkage and derangement. Immu-nohistochemical CD133 staining revealed that tumor cytoplasm color was brown. Conclusion Glioma model can be established based on glioma stem cells into a high rate of tumor, the tumor cycle is short, which can be used as an ideal model for glioma.
ABSTRACT
Objective To establish nude mouse model with human brain glioma SHG-44 and understand its growing characteristics in vivo.Methods The 4-week-old male mice were randomly divided into high density cell suspension inoculation group(n=10),low density cell suspension group(n=10),the tumor tissue mass vaccination group(n=10)and the blank control group with normal saline injection(n=10).The SHG-44 human brain glioma cell suspension was injected into the subcutaneous of the nude mice' s armpit.The tumor tissue was cut into 1 mm3 after tumor tissue growth and formation,and re-inoculated into the subcutaneous of the new nude mice' s armpits.Apart from daily observation,the long and short diameters of tumor were recorded every 5 days after graft.All the mice were sacrificed at 60 days and the tumor tissues were harvested for pathological examination.Results With a longer incubation period and slower growth rate,the tumor formation rate in high density cell suspension inoculation group and low density cell suspension group was lower compared with that in the tumor tissue mass vaccination group.Around day 20,grafted tumor appeared remarkably big((41.51 ±6.42)mm3) with good morphology.On day 50,the tumor derived from group the tumor tissue mass vaccination group((565.69± 123.36)mm3) showed a bigger size in comparison with that from high density cell suspension inoculation group((203.85±104.63) mm3) and low density cell suspension group ((153.02± 31.76) mm3,all P＜0.05).The tumors in three groups were well defined with a rich vascularity and no apparent invasion was observed.The positive expression of GFAP and S-100 in a large body of tumor cells was observed under optical microscope.Conclusion With a shorter incubation period and faster growth,the mouse tumor models established with tissue pieces from the tumor-bearing mice are much better compared to those with cell suspension.
ABSTRACT
Aim To observe the effect and primary mechanism of arctigenin ( ARG) in C6 rat glioma. At the same time, to investigate the effect of ARG com-bined with temozolomide. Methods C6 glioma rat model was established, and 90 rats were divided into six groups, which were subcutaneously administered with model, low and high ARG (0. 05 and 0. 1 mg· kg-1 , sc) , temozolomide (20 mg·kg-1 , p. o. ) , low ARG combined with temozolomide(TMZ / ARG 0. 05) and high ARG combined with temozolomide ( TMZ /ARG 0. 1 ) . The tumor specimens of brain were col-lected after tumor graft. Proliferating cell nuclear anti-gen ( PCNA ) , glial fibrillary acidic protein ( GFAP ) and CD40 in tumor specimens were determined by im-munohistochemistry. Results ① Compared with the model group, the tumor sizes of rats in the arctigenin treatment groups were decreased ( P significantly decreased PCNA and CD40 expression ( P<0. 05 ) and increased GFAP expression ( P<0. 05 ) .③ Compared with model group, arctigenin combined with temozolomide decreased the tumor sizes ( P <0. 01 ) , and the tumor inhibition rate was higher than that of the arctigenin and temozolomide. At the same time, arctigenin combined with temozolomide de-creased PCNA and CD40 expression ( P <0. 01 ) and increased GFAP expression ( P <0. 05 ) , which was better than arctigenin and temozolomide. Conclusion Arctigenin inhibits rat glioma growth, and synergizes with temozolomide, which may be associated with in-hibiting PCNA and CD40 expression and strengthening GFAP expression.
ABSTRACT
Objective:To evaluate the application value of diffusion tensor imaging (DTI) in guiding the postoperative radiothera-py plan of the gliomas adjacent to the corticospinal tract (CST). Methods:Thirty patients with gliomas adjacent to the CST underwent routine magnetic resonance imaging (MRI) contrast-enhanced scanning and DTI after radiotherapy. Tractography data sets were ac-quired and were fused with the images of corresponding anatomical MRI and computed tomography. The acquired data sets of radio-therapy planning system were imported to assist with the delineation of the target volume, organs at risk, and CST. Two sets of radio-therapy plan, which considered or did not consider the dose protective effect of the CST, were formulated and compared using the treat-ment technique of intensity modulated radiotherapy. Results:The protective radiotherapy and unprotected plans both achieved the thera-peutic dose to the target volume and the protection of the routine organs at risk. In the protective dose (with an optimization program that considered the dose reduction of CST), the maximum and mean radiation doses suffered by the patients' ipsilateral and contra-later-al CSTs were lower compared with the unprotected plan (P<0.05). Conclusion:DTI can identify the location and shape of CSTs, and their relationship with the postoperative radiotherapy target of gliomas. These findings contribute to the formulation of a protective ra-diotherapeutic regimen to keep the CST from the maximum and the mean radiation doses to the largest extent, thereby decreasing the possibility of nerve damage after radiotherapy.
ABSTRACT
OBJECTIVE: To develop a kind of multifunctional targeting epirubicin liposomes for the treatment of brain tumor, to characterize their physicochemical properties, and to observe their targeting effects on the brain microvascular endothelial cells (BM-VECs) and on the brain glioma cells. METHODS: The 2-amino-2-deoxy-β-Z)-glucopyranose(NH2-Glu) was used as a targeting molecule and conjugated with a cholesterol-polyethylene glycol derivative (Chol-PEG2000-NHS) for obtaining the targeting functional material aimed at targeting to glucose transporter-1 (Glut-1) on the BMVECs of blood-brain barrier and further targeting to the glioma cells. To prepare the multifunctional targeting epirubicin liposomes, the targeting functional material was modified onto the surface of liposomes, and epirubicin was loaded into the core of liposomes as the anticancer drug. The encapsulation efficiency, particle size, polydispersity indexes and Zeta potential of the liposomes were measured, their cellular uptakes were performed on the BMVECs and the glioma cells. The inhibitory effect was performed on the glioma cells. RESULTS: The analysis by MALDI-TOF-MS demonstrated that the targeting functional material, Chol-PEG2000-Glu, was successfully synthesized. The multifunctional targeting epirubicin liposomes were prepared, and had an average particle size of approximately 125 nm, and were negatively charged. The encapsulation efficiency of epirubicin in the liposomes was about 93%. Results from flow cytometry indicated that the multifunctional targeting epirubicin liposomes had the highest cellular uptakes by BMVECs and by two kinds of brain glioma cells as compared with no-targeting epirubicin liposomes. The cytotoxic study showed that the multifunctional targeting epirubicin liposomes had the strongest inhibitory effect to brain glioma cells as compared with free epirubicin or no-targeting epirubicin liposomes. CONCLUSION: A new targeting material (Chol-PEG2000-Glu) and the multifunctional targeting epirubicin liposomes are developed, and the multifunctional targeting epirubicin liposomes exhibit the potential lortransporting across the blood-brain barrier (BBB), and selectively inhibiting the brain glioma cells.
ABSTRACT
OBJECTIVE: To prepare and evaluate the novel core-shell structural phospholipid-functionalized mesoporous silica nanoparticles (MSN-LP) modified with angiopep-2 (ANG-MSN-LP). METHODS: Mesoporous silica nanoparticles (MSN) was synthesized by the modified Stober method. MSN-PTX was prepared by saturated solution adsorption method. ANG-MSN-LP was developed by selfassembly and film hydration method. By using dialysis bag method to investigate the in vitro drug release characteristics and MTT method to investigate the cytotoxicity on HBMEC and C6 cells. The transport ability and effects on cell cycle of the carrier was investigated by the BBB monolayer model. RESULTS: MSN was synthesized with high specific surface area (SBET, 425 m · g-1), cumulative pore volume (Vp, 0.37 cm · g-1) and pore size(3.5 nm). PTX was highly encapsulated (drug loading efficiency up to 11.1%) into MSN. Results of in vitro release showed that about 75.5% of PTX released from ANG-MSN-LP-PTX after 48 h and burst release was effectively reduced compared with MSN-PTX or PTX solution, indicating pronounced sustained-release characteristics. The good biocompatibility and low toxicity of ANG-MSN-LP were evaluated by HBMEC and C6 cells. The transport ratio was 2.49% for PTX, 2.72% for MSN-PTX, 4.45% for MSN-LP-PTX and 10.74% for ANG-MSN-LP-PTX respectively. In addition, ANG-MSN-LP-PTX showed a higher cell number of G2-M phase of 40.92 ± 6.20%. CONCLUSION: ANG-MSN-LP is a prospective targeting drug delivery system for therapy of brain glioma. Meanwhile, saturated solution adsorption method can increase the drug loading efficiency highly.
ABSTRACT
Objective To study the expression of transforming growth factor beta1(TGF-β1)and its type I receptors activin-like kinase 1(ALK1)in the human brain glioma and estimate the significance in the pathogenisis and development of human glioma . Methods The mRNA and protein expressions of TGF-β1 and ALK1 were detected with semiquantitative RT-PCR ,Western blot and Immunohistochemistry in 3 normal brain tissues and 32 patiens with human glioma .Results ALK1 and TGF-β1 mRNA and protein sometimes had co-expression in human gliomas tissues .Compared with those normal brain tissues ,the expression of TGF-β1 and ALK1 mRNA and protein in high grade gliomas were significantly increased (P<0 .05) ,the overexpression of ALK1 in human gliomas was demonstrated significant positive correlation with the pathological grades of gliomas (r=0 .297 ,P<0 .05) .Conclusion The overexpression of ALK1 may play important role in the development of human brain glioma .