1.Effects of morusin on the migration and invasion of U87 cells
Dong TANG ; Wenhong GAO ; Huaping ZHANG ; Qianxue CHEN
Practical Oncology Journal 2024;38(3):168-172
Objective The aim of this study was to investigate the effects of morusin on the migration and invasion of human glioblastoma U87 cells,and to explore its mechanism of action.Methods The Cell Counting Kit-8(CCK-8)was used to detect the effect of morusin on proliferation of U87 cells.Wound-healing and Transwell assays were used to detect the effects of morusin on mi-gration and invasion of U87 cells.Western blot was employed to detect the effect of morusin on the expressions of MMP-2 and Vimen-tin in U87 cells.Results The results of CCK-8 assay showed that morusin(1,2,4,and 6μg/mL)could significantly inhibit the pro-liferation of U87 cells compared to the 0 μg/mL morusin(P<0.001).The results of wound-healing assay showed that the migration rates of morusin-treated groups(2,4,and 6 μg/mL)were(20.597±1.225)%,(14.734±1.528)%and(7.811±1.496)%,respec-tively,which were significant lower than that in the 0 μg/mL group(40.566±3.284)%(P<0.001).The results of Transwell assay showed that the invasion number of U87 cells treated with morusin at the concentrations of 2,4,and 6 μg/mL was 85.000±6.557,41.000±6.245,and 13.333±3.215,respectively,which were significant lower than that in the 0 μg/mL group(116.667±14.572)(P<0.001).The results of Western blot showed that the expression of Vimentin in U87 cells increased gradually accompanying with the increase of morusin concentrations,while the expression of MMP-2 decreased gradually accompanying with the increase of morusin concentrations(P<0.001).Conclusion Morusin can effectively inhibit the migration and invasion ability of U87 cells,and its effect is positively correlated with the concentration of morusin within a certain range.
2.Effects of over-expression of E2F transcription factor 1 on radiosensitivity glioma U251 cells
Conggang HUANG ; Ming LUO ; Qianxue CHEN ; Yuan WANG ; Qiaochun HUANG ; Jie ZHOU ; Xingwan WANG ; Zhihua LUO ; Faliang DUAN
Chinese Journal of Behavioral Medicine and Brain Science 2023;32(3):218-224
Objective:To investigate the effects of over-expression of E2F transcription factor 1 (E2F1) on proliferation, invasion, apoptosis and radiosensitivity of glioma cell U251.Methods:Real-time quantitative PCR (qRT-PCR) were used to detect the differential expression of E2F1 mRNA in glioma cells LN18, SW1088, U251 and normal brain glial cells. The stable over-expression of E2F1 plasmid was constructed and transfected into U251 cells. qRT-PCR and Western blot test were used to detect the expression of E2F1, pituitary tumor transforming gene 1(PTTG1), C-Myc, B-cell lymphoma-2 (Bcl-2), Bcl2-associated X (Bax) mRNA and protein expression in the control group and E2F1 over-expression group.U251 cells were divided into control group(no X-ray irradiation), irradiation group(6 Gy dose of X-ray), and irradiation + E2F1 over-expression group(transfected with E2F1 first, then irradiated by 6 Gy of X-ray). Cell proliferation ability was detected by cell counting Kit-8(CCK-8) cell viability detection reagent, and cell invasion and migration ability were detected by Transwell chamber. Apoptosis and cell cycle were detected by flow cytometry.GraphPad Prism 8.0 was used for data analysis.The statistical methods were one-way ANOVA and independent sample t-test. Results:qRT-PCR showed that there was statistical difference in the mRNA levels of E2F1( F=201.92, P<0.05) in different cell lines.The expression levels of E2F1 mRNA in LN18(4.04±0.29), SW1088(3.19±0.16)and U251(4.66±0.20) cells were higher than those in HEB(1.02±0.07)cells ( q=27.00, 19.40, 32.52, all P<0.05). After successfully constructing U251 cells with stable over-expression of E2F1 plasmid, qRT-PCR and Western blot detection results showed that: the mRNA and protein levels of E2F1, PTTG1, C-Myc and Bcl-2 in E2F1 over-expression group were higher than those in control group ( t=77.16, 57.88, 4.63, 51.13, 7.50, 70.85, 8.38, 48.81, all P<0.05). Bax mRNA(0.20±0.01) and protein(0.66±0.01) levels were lower than those in control group((1.00±0.02), (0.94±0.01)), and the differences were statistically significant ( t=1.74, 54.65, both P<0.05). After X-ray irradiation (6 Gy), CCK8 detection results showed: the proliferation ability of the three groups at 24, 48, 72 and 96 h were significantly different ( F=95.41, 187.53, 1 158.49, 7 883.78, all P<0.05). The proliferation capacity of the irradiation group were lower than those of the control group at 24, 48, 72 and 96 h ( q=19.51, 27.20, 66.60, 174.9, all P<0.05). The proliferation capacity of irradiation + E2F1 over-expression group at 24, 48, 72 and 96 h were higher than those of irradiation group ( q=10.63, 10.81, 21.11, 60.90, all P<0.05). Transwell assay results showed that there were significant differences in cell invasion and migration ability among the three groups ( F=315.38, 681.10, both P<0.05). The invasion and migration ability of cells in the irradiation group were lower than those in the control group ( q=35.09, 12.76, both P<0.05), and the invasion and migration ability of cells in the irradiation + E2F1 over-expression group were higher than those in the irradiation group ( q=52.06, 22.81, both P<0.05). Flow cytometry showed that there were significant differences in apoptosis rate and percentage of cells in each cycle among the three groups ( F=667.63, 3 213.30, 3 011.26, 861.98, all P<0.05). The percentage of the apoptosis rate, S phase and G2 phase cells in the irradiation group were higher than those in the control group ( q=51.10, 89.39, 51.82, all P<0.05), while the percentage of G1 phase cells in the irradiation group was lower than that in the control group ( q=141.2, P<0.05). The apoptosis rate and percentage of S phase and G2 phase cells in the irradiation + E2F1 over-expression group were lower than those in the irradiation group ( q=18.87, 41.42, 29.31, all P<0.05), while the number of G1 phase cells in the irradiation + E2F1 over-expression group was lower than that in the irradiation group ( q=70.73, P<0.05). Conclusion:Over-expression of E2F1 can reduce the radiosensitivity of glioma U251 cells by regulating the expression of mRNA and protein of genes related to cell cycle and apoptosis, and E2F1 may be involved in the radioresistance of glioma cells.
3.Research Progress of Diagnosis and Treatment of Glioblastoma
Cancer Research on Prevention and Treatment 2022;49(11):1103-1106
Glioblastoma is the most common primary malignant brain tumor. Despite large-scale researches have been carried out, the prognosis of glioblastoma is still not significantly improved. In recent years, with the application of multi-omics studies in glioblastoma, the understanding of glioblastoma has been deepened. The classification of glioblastoma has been revised; moreover, new therapeutic methods such as targeted therapy, immunotherapy and tumor treating fields have been developed on the basis of the standard therapy. This article reviews the recent progress in the diagnosis and treatment of glioblastoma.
4.Mining prognostic marker of glioma based on weighted gene co-expression network analysis
Chunyu ZHANG ; Liguo YE ; Long WANG ; Yinqiu TAN ; Fanen YUAN ; Ye TAO ; Qianxue CHEN ; Daofeng TIAN
Journal of Chinese Physician 2021;23(4):529-533
Objective:To identify effective biomarkers for glioma patients.Methods:The mRNA expression profiles of 464 glioma patients with complete clinical follow-up information were downloaded from the Chinese Glioma Genome Atlas (CGGA). Weighted gene co-expression network analysis (WGCNA) was used to identify gene modules related to World Health Organization (WHO) grading of glioma, and univariate and multivatiate Cox regression analysis were performed to identify gliomas survival-related genes.Results:In weighted gene co-expression analysis, the module Brown was significantly positively correlated with glioma WHO stage ( r=0.55, P<0.05). In univariate analysis, five genes (TAGLN2, IGFBP2, METTL7B, ARAP3, PLAT) that were most significantly associated with clinical prognosis were selected for multivariate survival analysis, and the prognosis model was established to calculate the risk score. The receiver operating characteristic curve (ROC) confirmed that the risk score had high accuracy in predicting the 1-, 3-, 5-year survival rate of glioma patients. The above survival analysis results were verified in the Cancer Genome Atlas (TCGA) database. Conclusions:We use mRNA expression profiles to establish prognostic markers for gliomas to assess the overall survival of patients with glioma.
5.Effecacy of drilling and drainage combined with atorvastatin calcium tablets in treatment of chronic subdural hematoma
Chao LIU ; Mingchang LI ; Junmin WANG ; Changlin ZOU ; Yinqiu TAN ; Pian GONG ; Qi TIAN ; Qianxue CHEN
Chinese Journal of Neuromedicine 2018;17(3):295-300
Objective To observe the clinical efficacy of drilling and drainage combined with atorvastatin calcium tablets in treatment of chronic subdural hematoma (CSDH).Methods Totally,46 patients with CSDH,admitted to and received therapy in our hospital from January 2014 to January 2017,were selected for this research.These patients were divided into control group (n=16) and experimental group (n=30) according to therapeutic schemes.The patients from the control group underwent drilling and drainage.Besides that,the patients from the experimental group were given atorvastatin calcium tablets additionally,20 mg/d×2 months.Two months after that,the curative efficacy,hematoma volume before and after operation,pneumocephalus volume one week after operation,duration of tube drainage,length of hospital stay,China stroke scale (CSS) scores,activities of daily life-Barthel index scale (ADL-BI) and visual analog scale (VAS) score were compared between the patients from the two groups.Results Two months after treatment,patients from the experimental group had significantly decreased hematoma volume as compared with those from the control group (P<0.05).The hematoma volume in both groups 2 months after treatment was significantly decreased as compared with that before treatment (P<0.05).The pneumocephalus volume,indwelling time of drainage tube,and hospital stays in the experimental group were significantly shorter/lower than those in the control group (P<0.05).The CSS scores and VAS scores in the experimental group 2 months after treatment were significantly lower than those in the control group (P<0.05).The ADL-BI scores in the experimental group 2 months after treatment were significantly higher than those in the control group (P<0.05).The ADL-BI scores in both groups 2 months after treatment was significantly increased as compared with those before treatment (P<0.05).Conclusion As compared with simple use of drilling and drainage,drilling and drainage combined with atorvastatin calcium tablets can help hematoma absorption,decrease incidence of pneumocephalu,and improve prognosis effectively.
6.Predictive value of neuron-specific enolase in patients with mild cognitive impairment secondary to mild, moderate craniocerebral injury
Conggang HUANG ; Faliang DUAN ; Jinglei WU ; Qianxue CHEN ; Yuan WANG ; Ping SONG
Chinese Journal of Neuromedicine 2017;16(5):513-518
Objective To explore the predictive value of neuron-specific enolase (NSE) in patients with mild cognitive impairment (MCI) secondary to mild,moderate craniocerebral injury.Methods Seventy-six patients with mild,moderate craniocerebral injury,admitted to our hospital from March 19,2014 to September 1,2015,were chosen in our study;16 of them had secondary MCI during follow up (experimental group) and 60 did not show cognitive dysfunction (control group).Their clinical data between the two groups were compared.The predictive value of different NSE levels for secondary MCI was analyzed.Logistic regression analysis was performed to analyze the risk of secondary MCI in patients with different NSE levels.Multiple linear regression analysis was used to analyze the influence of mini-mental state examination (MMSE) scores in cognitive level of secondary MCI patients.Results (1) There were significant differences in age,salvage time,proportion of hypertension,ratio of skull fracture,injury severity scale (ISS) scores and total cholesterol (TC) between the experimental group and control group (P<0.05).(2) A lowest quartile group,second quartile group,third quartile group and highest quartile group were divided using NSE levels as independent variables (9.31 ng/mL-12.08 ng/mL,12.09 ng/mL-15.68 ng/mL,15.69 ng/mL-19.65 ng/mL and 19.66 ng/mL-23.47 ng/mL);following the increase of NSE levels,the age,salvage time,proportion of hypertension,and ISS scores were significantly increased (P<0.05);Single-factor and multivariate Logistic regression analyses showed that the risk of secondary MCI in the highest quartile group was 1.42 and 1.21 folds,respectively,as compared with that in the lowest quartile group.(3) Multiple linear regression analysis showed that baseline NSE level,age,salvage time,and ISS scores were the nfluence factors of MMSE scores in patients with secondary MCI;when the NSE content increased 1 ng/mL,MMSE decreased 0.369 points.Conclusion NSE level in patients with traumatic brain injury is an independent risk factor for secondary MCI,and its level is significantly associated with cognitive impairment.
7.Homer1a protein expression and its relationship with neuronal deficit and neuronal apoptosis in craniocerebral trauma patients
Conggang HUANG ; Faliang DUAN ; Jinglei WU ; Qiaochun HUANG ; Qiang MIN ; Ming LUO ; Qianxue CHEN ; Yuan WANG ; Ping SONG ; Jiqiang LI
Chinese Journal of Neuromedicine 2017;16(6):595-598
Objective To study the Homer1a protein expression and its relationship with neurological deficit and neuronal apoptosis in craniocerebral trauma patients. Methods Forty-two craniocerebral trauma patients, admitted to our hospital from May 2012 to March 2016, were selected as craniocerebral trauma group; 50 healthy subjects accepted physical examination at the same period in our hospital were selected as normal control group (n=50). Immediately after admission, serum contents of Homer1a protein and nerve function damage indices (neurospecific estrogenase [NSE]), fatty acid binding protein [FABP], insulin-like growth factor [IGF-1], and S100B protein) were measured by enzyme linked immunosorbent assay (ELISA). Serum apoptotic indices (soluble apoptotic factor [(sFas)], sFas ligand [sFasL], and cell lymphoma-2 [Bcl-2]) were detected by radioimmunoassay. Results Immediately after admission, serum content of Homer1a protein content in craniocerebral trauma group ([113.27±12.19] pg/mL) was significantly higher than that in normal control group ([53.93±4.06] pg/mL, P<0.05); the median serum Homer1a protein level was 115.302 pg/mL, and according to this level, the patients from the craniocerebral trauma group were further divided into high Homer1a group and low Homer1a group. Serum NSE, FABP, S100B, sFas and sFasL levels in the high Homer1a group, low Homer1a group and normal control group were decreased in sequence, and IGF-1 and Bcl-2 levels increased in sequence, with significant differences (P<0.05). Conclusion Expression of Homer1a protein is increased in patients with traumatic brain injury, and its content is directly related to nerve injury and neuron apoptosis.
8.Influence of intracranial pressure monitoring on GCS score,complications and prognosis of severe traumatic brain injury patients with trauma craniectomy
Journal of Clinical Medicine in Practice 2017;21(11):73-76
Objective To explore the influence of intracranial pressure monitoring on GCS score,complications and prognosis of severe traumatic brain injury patients with trauma craniectomy.Methods A total of 84 patients with STBI were randomly divided into two groups.The routine group was treated with standard large trauma craniectomy,and monitoring group was treated with trauma craniectomy combined with intracranial pressure monitoring.The change of intra-carnial pressure before and after operation,GCS scores change,prognosis as well as post operative complications were compared between two groups.Results Compared with routine group,the intra-carnial pressure at 3rd and 7th day after operation in monitoring group significantly decreased (P<0.05),and the GCS score at 28th day improved after operation in both groups,and increasing degree of monitoring group was significantly higher than the routine group (P<0.05).The GCS score at 3rd month after operation indicated that the prognosis in monitoring group was significantly better than routine group (P<0.05).The incidence rates of electrolyte disturbance in monitoring group and routine group at 6 month after surgery were 59.5% and 33.3% respectively (P<0.05).Conclusion Standard trauma craniectomy combined with intracranial pressure monitoring treatment is effective in treatment of patients with severe traumatic brain injury.
9.Influence of intracranial pressure monitoring on GCS score,complications and prognosis of severe traumatic brain injury patients with trauma craniectomy
Journal of Clinical Medicine in Practice 2017;21(11):73-76
Objective To explore the influence of intracranial pressure monitoring on GCS score,complications and prognosis of severe traumatic brain injury patients with trauma craniectomy.Methods A total of 84 patients with STBI were randomly divided into two groups.The routine group was treated with standard large trauma craniectomy,and monitoring group was treated with trauma craniectomy combined with intracranial pressure monitoring.The change of intra-carnial pressure before and after operation,GCS scores change,prognosis as well as post operative complications were compared between two groups.Results Compared with routine group,the intra-carnial pressure at 3rd and 7th day after operation in monitoring group significantly decreased (P<0.05),and the GCS score at 28th day improved after operation in both groups,and increasing degree of monitoring group was significantly higher than the routine group (P<0.05).The GCS score at 3rd month after operation indicated that the prognosis in monitoring group was significantly better than routine group (P<0.05).The incidence rates of electrolyte disturbance in monitoring group and routine group at 6 month after surgery were 59.5% and 33.3% respectively (P<0.05).Conclusion Standard trauma craniectomy combined with intracranial pressure monitoring treatment is effective in treatment of patients with severe traumatic brain injury.

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