Construction of a pyroptosis-related lncRNAs signature for predicting prognosis and immunotherapy response in glioma

Recent studies have proved that pyroptosis-related long non-coding RNAs (PRlncRNAs) are closely linked to tumor progression, prognosis, and immunity. Here, we systematically evaluated the correlation of PRlncRNAs with glioma prognosis. This study included 3 glioma cohorts (The Cancer Genome Atlas, Chinese Glioma Genome Atlas, and Gravendeel). Through Pearson correlation analysis, PRlncRNAs were screened from these 3 cohorts. Univariate Cox regression analysis was then carried out to determine the prognostic PRlncRNAs. A pyroptosis-related lncRNAs signature (PRLS) was then built by least absolute shrinkage and selection operator and multivariate Cox analyses. We systematically evaluated the correlation of the PRLS with the prognosis, immune features, and tumor mutation burden in glioma. A total of 14 prognostic PRlncRNAs overlapped in all cohorts and were selected as candidate lncRNAs. Based on The Cancer Genome Atlas cohort, a PRLS containing 7 PRlncRNAs was built. In all cohorts, the PRLS was proved to be a good predictor of glioma prognosis, with a higher risk score related to a poorer prognosis. We observed obvious differences in the immune microenvironment, immune cell infiltration level, and immune checkpoint expression in low- and high-risk subgroups. Compared with low-risk cases, high-risk cases had lower Tumor Immune Dysfunction and Exclusion scores and greater tumor mutation burden, indicating that high-risk cases can be more sensitive to immunotherapy. A nomogram combining PRLS and clinical parameters was constructed, which showed more robust and accurate predictive power. In conclusion, the PRLS is a potentially useful indicator for predicting prognosis and response to immunotherapy in glioma. Our findings may provide a useful insight into clinically individualized treatment strategies for patients.     

Dickkopf1在人脑胶质瘤细胞株中的表达及意义

目的探讨Wnt通路抑制因子Dickkopf-1（DKK1）在人胶质瘤发生、发展中的作用。方法采用ELISA、RT-PCR法检测人胶质瘤细胞株U251和T98G中DKK1蛋白及DKK1 mRNA表达水平。结果DKK1蛋白在U251和T98G中均呈高表达;DKK1mRNA在胶质瘤细胞株U251和T98G中高表达,但在正常脑组织中表达不明显。结论DKK1在人胶质瘤细胞株中高表达;其可能在胶质瘤发生发展中起重要作用。     

FHIT蛋白在人脑胶质瘤组织中的表达及临床意义

目的 探讨人脑胶质瘤组织脆性组胺三联体（FHIT）的表达及其临床意义。方法 应用免疫组织化学SP法,检测38例人脑胶质瘤组织（观察组;低度恶性13例。高度恶性25例）和8例正常脑组织（对照组）中FHIT蛋白的表达。结果 观察组FHIT蛋白表达率（47．4%）明显低于对照组（100％）,P〈0．01;低度恶性者FHIT蛋白表达率（53．8％）明显高于高度恶化者（44．0％）,P〈0．05。结论 FHIT的异常表达与人脑胶质瘤的发生、发展有关。     

氯化镧诱导脑胶质瘤SHG44细胞凋亡作用的机制

目的探讨氯化镧对人脑胶质瘤细胞株SHG44细胞诱导凋亡及其作用机制。方法体外培养人胶质瘤细胞,流式细胞仪、电镜检查细胞周期和超微结构的改变,原位杂交技术及免疫组化染色技术检测bcl-2、bax基因、蛋白表达情况。结果氯化镧能促进入脑胶质瘤SHG44细胞发生凋亡,明显增加G1期DNA含量,减少S期DNA含量。使bax基因、蛋白表达显著增强,bcl-2、bax基因蛋白表达显著减弱（P〈0．05）。结论氯化镧可抑制人脑胶质瘤SHG44细胞的生长,并促进其凋亡。     

u-PAR在人脑胶质瘤中的表达及意义

目的探讨尿激酶型纤溶酶原激活物受体（u-PAR）在人脑胶质瘤中的表达及其与临床病理分级的关系。方法应用流式细胞仪定量检测35例人脑胶质瘤标本中u-PAR水平,其中Ⅰ级8例,Ⅱ级9例,Ⅲ级9例,Ⅳ级6例,正常脑组织3例,应用直接染色法。结果u-PAR在人脑胶质瘤中的表达与病理分级相关,Ⅲ、Ⅳ级与Ⅰ、Ⅱ级和正常脑组织比较有高度统计学差异（P〈0.01）。结论u-PAR表达与人脑胶质瘤病理分级相关,可作为判断其恶性程度和浸润性能的指标。     

Notch1信号通路在人脑胶质瘤U87细胞增殖、凋亡中的作用及机制探讨

目的 观察Notch1信号通路在人脑胶质瘤U87细胞增殖和凋亡中的作用,并探讨其机制.方法 将U87细胞随机分为A、B、C组,分别加入Notch信号激活剂rhNF-κB、抑制剂DAPT及PBS共培养48 h.用MTT法检测细胞吸光度值(A值)观察细胞增殖情况,流式细胞术检测细胞凋亡率,RT-PCR、Western blot法分别检测U87细胞中的Notch1、Hes1、Bcl-2 mRNA及其蛋白.结果 培养24、48、72、96 h,A组细胞A值分别为0.185±0.007、0.398±0.012、0.735±0.015、1.083±0.031,B组分别为0.102±0.003、0.130±0.004、0.161±0.006、0.194±0.003,C组分别为0.167±0.005、0.265±0.008、0.496±0.011、0.737±0.016,A、B组与C组比较,P均＜0.05.A、B、C组细胞凋亡率分为0.96％±0.17％、26.51％±3.74％、8.76％±1.40％,A、B组与C组比较,P均＜0.05.与C组比较,A组细胞中Notch1、Hes1、Bcl-2 mRNA及其蛋白表达量均显著增加(P均＜0.05),B组细胞中Notch1、Hes1、Bcl-2 mRNA及其蛋白表达量均显著降低(P均＜0.05).结论 Notch1信号通路在人脑胶质瘤U87细胞增殖、凋亡中发挥了重要的调控作用,其机制可能与调节靶基因Hes1、抗凋亡蛋白Bcl-2表达有关.     

脑胶质瘤组织中MCM5、Ki-67的表达及临床意义

目的探讨微小染色体维持蛋白5（MCM5）、细胞增殖相关核蛋白（Ki-67）在脑胶质瘤发生、发展中的意义。方法选择手术切除的脑胶质瘤组织44份（肿瘤组）和正常脑组织6份（正常组）,采用免疫组化法检测两组MCM5、Ki-67表达情况[以标记指数（LI）表示],用Spearman等级相关分析法分析两指标问及与肿瘤组临床病理特征的相关性。结果肿瘤组、正常组MCM5的LI分别为12．964-8．49、0,P〈0．05;Ki-67的u分别为10．09±7．07、0,P〈0．0．5。MCM5、Ki-67阳性表达与脑胶质瘤恶性程度密切相关;MCM5与Ki-67表达呈正相关,在不同分级肿瘤中MCM5表达均显著高于Ki-67。结论MCM5、Ki-67高表达在脑胶质瘤发生、发展中起重要作用;MCM5反映肿瘤细胞的增殖状态更为灵敏。     

异硫氰酸苄酯对脑胶质瘤U-87MG细胞周期的影响及其机制

目的观察异硫氰酸苄酯（BITC）对人脑胶质瘤细胞系U一87MG细胞周期的阻滞作用,并探讨其机制。方法取对数期u．87MG细胞分别以2、5Ixmol／L的BITC处理,应用流式细胞仪检测细胞周期分布,Westernblot—ting法检测细胞周期相关蛋白CyclinB1、Cdc2、p21、Cdc25C及磷酸化Akt（p-Akt）蛋白表达,RT—PCR法检测CyclinB1、Cdc2、p21和Cdc25C的mRNA表达,报告基因技术检测NF—KB、缺氧诱导因子（HIF）和真核细胞翻译起始因子4E（eIF4E）的转录活性。结果BITC作用24h后,u一87MG细胞周期G2／M期比例显著升高,CyclinB1、Cdc2、Cdc25C、p-Akt蛋白表达下调,p21蛋白表达上升,CyclinB1、Cdc2、Cdc25C、p21mRNA表达均显著下降,NF—KB、HIF、elF4E转录活性均显著下降（P均〈0．05）。结论BITC对U．87MG细胞周期有阻滞作用,机制可能与抑制Akt／NF-KB信号转导路径,进而调节细胞周期相关基因表达有关;此为胶质瘤的细胞周期靶点治疗提供了新思路。     

High expression of microRNA 221 is a poor predictor for glioma

Background:MicroRNA 221 has been found to be a good marker for several cancers. Some studies also focused on the relationship between microRNA 221 and glioma. However, the results are controversial. We aimed to systematically evaluate the prognostic role of microRNA 221 in glioma through performing a meta-analysis.Methods:The articles which were included in our study were searched on the Web of Science, EMBASE, PubMed, Cochrane Library and China National Knowledge Infrastructure. The basic characteristics and relevant data were extracted. Hazard ratios (HRs) with 95% confidence intervals (CIs) were pooled to evaluate the prognostic role of microRNA 221 in glioma.Results:Eight studies with 1069 patients were included. We systematically evaluated the role of microRNA 221 for overall survival (OS) and disease free survival (DFS) in glioma patients (HR for OS = 1.66, 95% CI, 1.34–2.04; HR for DFS = 1.14, 95% CI, 1.02–1.26). Subgroup analyses were performed according to the nation of the studies, the origin of the samples, the stage of the tumors, the cut-off value, and the method for detecting the microRNA 221. No significant publication bias was found (P = .133).Conclusion:In conclusion, high expression of microRNA 221 was related to poor prognosis of glioma. These findings may assist future exploration on microRNA 221 and help predict the prognosis of glioma. However, due to the significant heterogeneity of these studies, more studies are warranted.     

人参皂甙单体Rh2对人脑胶质瘤细胞U251增殖和凋亡的影响及其机制探讨

目的观察人参皂甙单体Rh2（GS-Rh2）对人脑胶质瘤U251细胞增殖和凋亡的影响,并探讨其机制。方法用GS-Rh2处理人脑胶质瘤U251细胞,采用MTT法检测U251细胞增殖抑制率,流式细胞仪和Annexin V-FITC/PI双染法观察U251细胞凋亡情况,TRAP-ELISA法检测U251细胞端粒酶活性,RT-PCR法检测U251细胞人端粒酶逆转录酶（hTERT）mRNA的表达。结果 5、10、20、40、80μg/ml GS-Rh2组U251细胞增殖抑制率分别为20.42%、32.19%、45.09%、57.37%、73.82%。根据细胞增殖抑制曲线计算出IC30、IC50、IC70值分别为9.7、25.2、68.4μg/ml。FITC和PI荧光双参数点图显示实验组细胞分布区域与对照组相比出现明显改变,随着药物浓度的增加,早期凋亡、晚期凋亡或坏死的区域细胞数量逐渐增多。9.7、25.2、68.4μg/ml GS-Rh2处理12、24、48、72 h后U251细胞端粒酶活性随GS-Rh2浓度增加和作用时间的延长而降低。浓度为25.2μg/ml GS-Rh2作用24、48、72 h的U251细胞hTERTmRNA与β-actin灰度值比为0.964 9±0.004 2、0.401 8±0.001 4、0.297 8±0.001 8,对照组为0.976 4±0.005 1。作用48、72 h U251细胞hTERT mRNA与β-actin灰度值比与对照组相比P均〈0.05;作用48、72 h者相比P〈0.05。结论 GS-Rh2能够诱导人脑胶质瘤U25l细胞凋亡,抑制U25l细胞增殖。其机制与其抑制hTERT基因表达,降低端粒酶活性有关。     

DAPT suppresses the proliferation of human glioma cell line SHG-44

Objective:To explore the suppressing effect ofγ-secretase inhibitor DAPT on proliferation of human glioma cell line SHG-44 in vitro and its mechanism.Methods:The SHG-44 cell was treated by DAPT with different concentration.The proliferation of cells was detected by MTT assay;cell cycle and TSC of CD133~+were determined by flow cytometry analysis technique;the key factor in Notch signaling pathway(Notch-1,Delta-1,Hes-1)was measured by reverse transcrip tase-polymerase chain reaction and western blotting.Results:DAPT inhibited the growth and proliferation of SHG-44 cells significantly(P0.05).And the inhibiting effect on SHG-44 cells produced by DAPT showed a dose-dependent manner.DAPT increased the rate of cells in G_0/G_1 phase of SHG-44 cells,while it decreased the rate of cells in S phase.TSC of CD133~+was significantly reduced after DAPT treated SHC-44 cells.The expression of protein and mRNA of Notch-1,Delta-1 and Hes-1 were gradually downregulated with the increase of DAPT doses.Conclusions:DAPT can downregulate these key factor in Notch signaling pathway,reduce the TSC of CD133+and inhibit the proliferation of SHC-44 cells.     

Dandruff-associated Malassezia genomes reveal convergent and divergent virulence traits shared with plant and human fungal pathogens

Fungi in the genus Malassezia are ubiquitous skin residents of humans and other warm-blooded animals. Malassezia are involved in disorders including dandruff and seborrheic dermatitis, which together affect >50% of humans. Despite the importance of Malassezia in common skin diseases, remarkably little is known at the molecular level. We describe the genome, secretory proteome, and expression of selected genes of Malassezia globosa. Further, we report a comparative survey of the genome and secretory proteome of Malassezia restricta, a close relative implicated in similar skin disorders. Adaptation to the skin environment and associated pathogenicity may be due to unique metabolic limitations and capabilities. For example, the lipid dependence of M. globosa can be explained by the apparent absence of a fatty acid synthase gene. The inability to synthesize fatty acids may be complemented by the presence of multiple secreted lipases to aid in harvesting host lipids. In addition, an abundance of genes encoding secreted hydrolases (e.g., lipases, phospholipases, aspartyl proteases, and acid sphingomyelinases) was found in the M. globosa genome. In contrast, the phylogenetically closely related plant pathogen Ustilago maydis encodes a different arsenal of extracellular hydrolases with more copies of glycosyl hydrolase genes. M. globosa shares a similar arsenal of extracellular hydrolases with the phylogenetically distant human pathogen, Candida albicans, which occupies a similar niche, indicating the importance of host-specific adaptation. The M. globosa genome sequence also revealed the presence of mating-type genes, providing an indication that Malassezia may be capable of sex.     

AAV—AS联合环磷酰胺治疗大鼠皮下胶质瘤效果观察

目的观察腺相关病毒为载体血管抑素重组质粒（AAV-AS）联合环磷酰胺（CTX）治疗大鼠皮下胶质瘤的效果。方法大鼠皮下抑制胶质瘤C6细胞,建立荷C6胶质瘤大鼠模型,当肿瘤体积达250～280mm3时,随机分为对照组、CTX组、AAV-AS组及联合组,分别腹腔内注射生理盐水、CTX、AAV-AS、CTX＋从V—AS。每隔3d测量肿瘤的长、短径计算体积;于22d后处死大鼠,采用免疫组化染色法标记抗CD31抗体计算瘤组织中的微血管密度（MVD）,TUNEL法测算肿瘤细胞凋亡指数（AI）。结果对照组肿瘤体积为（2060．339±82．028）mm3,MVD为（20±3）条／HP,AI为1．442±0．329;CTX组分别为（1763．867±124．237）mm3、（18±5）条／HP、1．719±0．464,AAV—AS组分别为（1229．910±141．646）mm3、（13±2）条／HP、3．653±0．238,联合组分别为（1098．320±70．880）mm3、（8±3）条／HP、3．886±0．361。联合组肿瘤体积和MVD与其他组比较,P均〈0．05;联合组AI与对照组及CTX组比较,P均〈0．05,但与AAV—AS组比较,P〉0．05。结论AAV-AS联合CTX治疗大鼠皮下胶质瘤,可减小肿瘤体积,降低血管密度,间接促进肿瘤细胞凋亡。     

Antiproliferative activity of the non-myelotoxic antitumour agent of plant origin,Thaliblastine, on two human glioma cell lines

Summary The antiproliferative activity of the non-myelotoxic antitumour agent of plant origin, Thaliblastine, on two human glioma cell lines is described. Thaliblastine was added once one day following start of culture; proliferation was monitored over 7 days. The anti-proliferative activity of Thaliblastine was strongly dependent on concentration and time of incubation. The ID₅₀ of Thaliblastine in T406 and GW27 glioma lines was 5.1 g/ml and 8.2 g/ml (7.0 M and 11.2 M), respectively.Abbreviation TBL Thaliblastine     

地塞米松对胶质瘤C6细胞增殖、凋亡的影响

目的 观察地塞米松(Dex)对胶质瘤C6细胞增殖和凋亡的影响.方法 将C6细胞分为A、B、C、D组,每组1.25×106个细胞,A组不加Dex,B、C、D组分别与终浓度为10-5、10-4、10-3 mol/L的Dex共培养;用血计数板计算C6细胞数,用流式细胞仪检测C6细胞周期及凋亡率.结果 培养24 h时,A、B、C、D组C6细胞数依次为4.37×106、4.29×106、3.57×106、3.44 ×106个/瓶,B、C、D组与A组比较,P均＜0.05;C6细胞凋亡率依次为0.37％、0.52％、1.39％、8.24％,D组与A、B、C组比较,P均＜0.05.G0/G1期C6细胞所占比例分别为82.42％、93.21％、93.71％、77.52％,S期分别为13.22％、5.38％、4.06％、14.74％,G2/M期分别为4.36％、1.41％、2.23％、7.74％;B、C组与A组G0/G1、S期细胞比例比较,P均＜0.05;D组与A、B、C组G2/M期细胞比例比较,P均＜0.05.结论 Dex可通过阻滞C6细胞周期进程抑制细胞增殖、诱导细胞凋亡.     

丙戊酸对胶质瘤细胞化疗敏感性的影响

目的探讨丙戊酸（VPA）体外对胶质瘤细胞株化疗敏感性的影响,进一步提高胶质瘤的化疗效果。方法采用MTT法检测顺铂（DDP）和替莫唑胺（TMZ）单独应用及分别与1.0mmol/LVPA联合应用后胶质瘤细胞的细胞增殖率,计算半数抑制浓度（IC50）及增敏倍数（ER）。结果与单纯DDP处理者比较,VPA联合DDP处理后胶质瘤细胞株SF295、SKMG-1、UW28、SF767IC50显著降低（P〈0.05）;与单纯TMZ处理者比较,VPA联合TMZ处理后胶质瘤细胞株U87、UWR7、UW28、SF767IC50显著降低（P〈0.05）。结论 VPA能够增强部分胶质瘤细胞株对化疗药物的敏感性。     

EFNA1 is a potential key gene that correlates with immune infiltration in low-grade glioma

EFNA1 is a key gene that is associated with the pathogenesis of several human cancers. However, the prognostic role of EFNA1 in many cancers and the relationship between EFNA1 and tumor-infiltrating lymphocytes in different cancers remain unclear.The expression levels of EFNA1 in 33 types of cancer in the TCGA (The Cancer Genome Atlas) database were collected via the UCSC Xena browser. The clinical data of LGG (low grade glioma) patients were downloaded from the TCGA database. The glioma data from the CGGA (Chinese Glioma Genome Atlas) database were also downloaded to verify the results. Kaplan–Meier and Cox regression analyses were used to investigate the prognostic value of EFNA1 in different cancers using R software. We verified the differential expression of EFNA1 in glioma and normal brain tissue via gene expression profiling interactive analysis. We evaluated the relationship between the expression level of EFNA1 and the clinicopathological features of LGG patients via the Wilcoxon signed-rank test. The immune infiltration levels were evaluated via tumor immune estimation resource (TIMER) and CIBERSORT, and the correlations between EFNA1 and immune cell levels were investigated via TIMER. Finally, we conducted gene set enrichment analysis (GSEA) to explore the potential mechanisms.Data from the TCGA database showed that EFNA1 was differentially expressed in many kinds of cancers when compared with normal tissues. Upregulated EFNA1 expression in esophageal carcinoma (ESCA), cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC), and LGG correlated with shorter patient overall survival (OS) times. The Cox regression analysis revealed that the expression of EFNA1 was also a risk factor for the disease-specific survival (DSS) and progression-free interval (PFI) of LGG patients. The multiple Cox regression analysis revealed that EFNA1 was an independent prognostic factor for LGG patients. In addition, EFNA1 expression was increased in the WHO grade III group and the 1p19q non-codeletion group. Moreover, EFNA1 expression was positively correlated with the levels of infiltrating CD4+ T cells, myeloid dendritic cells and neutrophils in LGG. GSEA suggested that several GO and kyoto encyclopedia of genes and genomes (KEGG) items associated with nervous system function and apoptotic pathway were significantly enriched in the EFNA1-low and EFNA1-high expression phenotypes.EFNA1 may play a pivotal role in the development of LGG and may serve as a potential marker for LGG prognosis and therapy.     

胶质瘤手术患者阿霉素局部化疗的并发症分析

目的观察局部应用阿霉素治疗胶质瘤患者的并发症及其预防措施。方法对手术治疗的32例胶质瘤患者手术灶处铺垫含有阿霉素2.5-10 mg的明胶海棉,并于瘤腔内留置ommaya化疗泵导管。术后经ommaya泵反复注入阿霉素2.5-5 mg/次,同时放疗者21例。结果 32例手术患者局部化疗2、3、4、5、6次者分别为9、9、7、2、3例,共122次。并发脑内出血5例次、癫痫3例次、颅内压增高并头痛3例7次。颅内出血者均同时合并放疗,颅内压增高并头痛者单次用阿霉素5 mg者5次、10 mg者2次。结论手术治疗胶质瘤患者行阿霉素局部化疗时可发生脑内出血、癫痫、头痛等并发症。调整单次阿霉素用量、避免同时放疗、减慢输注速度等可以避免这些并发症的发生。