目的 分析脑胶质瘤中黑色素瘤相关抗原(MAGE)-D4和肌动蛋白结合蛋白CORO1C mRNA的表达水平,探讨二者的相关性及临床意义.方法 通过基因表达谱交互式分析(GEPIA)数据库收集脑胶质瘤样本数据681例[低级别脑胶质瘤(LGG)518例,胶质母细胞瘤(GBM)163例],正常脑组织样本数据207例,比较其MAGE-D4和CORO1C mRNA的表达水平.进一步收集62例脑胶质瘤组织和11例正常脑组织临床样本,应用逆转录-定量聚合酶链反应(RT-qPCR)检测其MAGE-D4和CORO1C mRNA表达水平并进行比较,分析MAGE-D4和CORO1C mRNA表达水平与脑胶质瘤患者临床指标的关联性.应用小干扰RNA(siRNA)技术沉默脑胶质瘤细胞株MAGE-D4的表达,检测CORO1C mRNA的表达变化情况.结果 GEPIA数据库数据分析结果显示,GBM和LGG组织的MAGE-D4和CORO1C mRNA表达水平均高于正常脑组织,差异有统计学意义(P<0.05).针对临床样本的RT-qPCR检测结果也显示,脑胶质瘤组织中MAGE-D4和CORO1C mRNA的表达水平显著高于正常脑组织(P<0.05).MAGE-D4及CORO1C mRNA的表达水平与患者年龄、性别、世界卫生组织(WHO)分级、病理类型、卡氏评分(KPS)、肿瘤最长径,以及Ki-67、胶质纤维酸性蛋白(GFAP)、S-100和波形蛋白(Vimentin)的表达水平均无显著关联(P>0.05).Pearson相关分析结果显示,临床脑胶质瘤样本组织中MAGE-D4与CORO1C mRNA表达水平呈正相关(r=0.848,P=0.000).细胞实验结果显示,经MAGE-D4 siRNA沉默后,U87 MG和U251细胞CORO1C mRNA的表达水平显著下降(P<0.05).结论 MAGE-D4和CORO1C mRNA在脑胶质瘤组织中高表达,且二者表达水平呈正相关.MAGE-D4和CORO1C可能协同影响脑胶质瘤的发生、发展过程,其具有成为脑胶质瘤分子靶向治疗靶点的潜力.

目的 探讨癌?睾丸抗原NY?SAR?35在结直肠癌(CRC)组织中的表达情况及其临床意义.方法 选择2009年10月至2012年5月于广西医科大学第一附属医院普外科接受手术治疗的CRC患者59例.男42例,女17例;年龄30～86(55.55±14.36)岁;TNM分期为Ⅰ～Ⅱ期38例,Ⅲ～Ⅳ期21例.均经手术取癌组织及配对的癌旁组织.采用逆转录聚合酶链反应(RT?PCR)检测组织NY?SAR?35 mRNA的表达情况.分析CRC组织NY?SAR?35 mRNA表达情况与患者临床特征及预后的关联性.结果 CRC组织中NY?SAR?35 mRNA阳性表达率显著高于癌旁组织(35.59％vs 13.56％,P<0.05).CRC组织NY?SAR?35 mRNA阳性组肿瘤直径≥5 cm的人数比例显著大于阴性组(P<0.05),但两组在性别、年龄、肿瘤位置、癌胚抗原(CEA)水平、T分期、N分期、M分期、TNM分期、组织学类型和分化程度方面比较差异无统计学意义(P>0.05).NY?SAR?35 mRNA阴性组的生存预后优于阳性组(生存期:57.24个月vs 40.71个月;log?rank检验:χ2=4.356,P=0.037).Cox回归分析结果显示,T分期为T3～4期、N分期为N1～3期、M分期为M1期以及NY?SAR?35 mRNA阳性表达是影响CRC患者生存预后的独立危险因素(P<0.05).结论 NY?SAR?35 mRNA在CRC组织中高表达,且与患者不良预后具有相关性,具有作为CRC 治疗靶点的潜在价值.

Glioma is the most common malignant brain tumor in central nervous system. Despite advances in the treatment of glioma such as surgery and chemoradiotherapy, most patients are easy to relapse, resulting in adverse clinical outcomes. Hence, effective molecular-targeting treatment may be one of attractive strategies for glioma therapy. The dysregulated microRNAs (miRNAs), one of the candidates of therapeutic targets, are believed to play an important role in the progression of glioma. In this study, we aimed to examine the expression profile of miRNAs in glioma and provide a reference for glioma therapy. Firstly, expression profile of miRNAs in 5 normal brain tissues, 5 low-grade glioma (LGG) tissues and 5 glioblastoma (GBM) tissues was detected by RNA sequencing (RNA-seq). Next, the target genes of differentially expressed miRNAs (DEmiRNAs) were predicted and then GO enrichment and KEGG pathway analysis performed by bioinformatics. Finally, 10 miRNAs which were significantly up- or down-regulated both in GBM and LGG were validated by real-time quantitative PCR (qRT-PCR). RNA-seq results indicated a number of DEmiRNAs in glioma. There were 64 up-regulated miRNAs and 17 down-regulated miRNAs in LGG, and 181 up-regulated miRNAs and 124 down-regulated miRNAs in GBM, respectively. Bioinformatics analysis showed that the target genes of these DEmiRNAs were enriched in various biological processes and signaling pathways such as cell metabolic and developmental process. Selected DEmiRNAs were further confirmed by qRT-PCR. miRNA-10b-5p, miRNA-92b-3p and miRNA-455-5p were significantly up-regulated in both GBM and LGG; while miRNA-542-3p was significantly up-regulated in LGG; miRNA-184 and miRNA-206 were significantly down-regulated in both GBM and LGG; miRNA-766-5p and miRNA-1-3p were significantly down-regulated in GBM. The subject of our study demonstrated several dysregulated miRNAs may serve as a potential therapeutic target for glioma.

目的 探讨肿瘤相关抗原MAGE-D4在肝细胞癌(HCC)中的表达情况,并分析其临床意义.方法 基于癌症基因组图谱(TCGA)数据库,获取371例HCC组织和50例正常肝组织的转录数据.应用数据挖掘工具UALCAN分析HCC组织中MAGE-D4 mRNA的表达情况,并筛选与其表达相关的基因.应用Kaplan-Meier Plotter分析MAGE-D4 mRNA表达水平与患者生存预后的关系.另收集于广西医科大学第一附属医院2009年6月至2011年8月经手术获取的HCC标本70例,癌旁组织标本30例,采用免疫组织化学法(IHC)检测MAGE-D4蛋白的表达情况,并分析其与患者临床指标的关联性.结果 基于TCGA数据库数据的分析结果显示,MAGE-D4 mRNA在HCC组织中的表达水平显著高于正常肝组织(P<0.05),且与甲胎蛋白(AFP)mRNA表达呈正相关(r=0.556,P=0.000).生存分析结果显示,MAGE-D4 mRNA高表达与黄种人HCC患者不良生存预后具有显著关联(HR=1.860,P=0.037).基于临床获取标本的分析结果显示,MAGE-D4蛋白主要定位于细胞质和细胞核,HCC组织的MAGE-D4高表达率显著大于癌旁组织(48.57％vs 23.33％;χ2=5.530,P=0.019).MAGE-D4高表达组HCC患者血清AFP浓度≥400μg/L的人数比例显著大于低表达组(P<0.05),但两组在性别、年龄、肿瘤级别、肿瘤分期、肿瘤直径、癌栓和乙型肝炎病毒(HBV)感染等方面比较差异无统计学意义(P>0.05).结论 MAGE-D4在HCC组织中高表达,其可能在HCC的发生和发展过程中发挥重要作用,有望成为HCC的潜在治疗靶点和预后标志物.

BACKGROUND The family of MAGE genes is well known due to the majority of MAGE genes expressing specifically in tumor tissues while restrictedly in normal tissues. MAGE-D4 is one of the MAGE family and considered as a promising target for glioma immunotherapy because of its overexpression in glioma and restricted expression in normal tissues. Whereas the mechanism of MAGE-D4 heterogeneous expression in glioma has not yet been elucidated. In this study, the transcriptional regulation mechanism of MAGE-D4 in glioma is focused from the perspectives of promoter methylation and SP1. METHODS Dual-luciferase reporter assay was performed to identify the core promoter of MAGE-D4 gene. Mass spectrometry was applied to quantify the methylation status of MAGE-D4 promoter in 50 glioma and 9 normal brain tissues. The influence of methylation and SP1 on MAGE-D4 transcriptional activity was evaluated by dual-luciferase reporter assay, qRT-PCR, western blot and ChIP-qPCR. Decitabine, an epigenetic drug, was used to treat the glioma cells. Then the treated cells were evaluated the influence of demethylation on SP1 binding to MAGE-D4 promoter. RESULTS The -358 to +172 bp region was identified as the core promoter of MAGE-D4 gene which demonstrated hypomethylated and negative correlation between methylation level and MAGE-D4 mRNA expression in glioma tissues. For single CpG unit analysis, 8 CpG units (CpG unit 1, 2, 3, 4, 5, 6, 9 and 12) in MAGE-D4 core promoter showed hypomethylated in glioma and the methylation level of CpG unit 6 was positively associated with the prognosis of glioma patients. Furthermore, the methylation level of CpG unit 1 and 6 was negative negatively correlated with MAGE-D4 mRNA expression. Then, the results demonstrated that the promoter activity of MAGE-D4 was decreased by methylation in glioma cell lines. In addition, SP1 can binds directly to the MAGE-D4 promoter leading to up-regulation of MAGE-D4 mRNA through activation of its promoter. Finally, demethylation of MAGE-D4 promoter could benefit the SP1 binding and resulting co-activation of MAGE-D4 promoter by demethylation and SP1 in glioma cell lines. CONCLUSION These findings indicate that the synergies of promoter hypomethylation and SP1 up-regulated MAGE-D4 transcription in glioma, which implies a potential approach to resolve the heterogeneous expression of MAGE-D4 in order to establish foundation for the MAGE-D4 based glioma therapy.

目的 检测肿瘤相关抗原M AGE-D4剪接变体ab和c在胶质瘤和非肿瘤脑组织中的表达情况,分析其临床意义,探讨M AGE-D4选择性剪接与胶质瘤的相关性.方法 采用RT-PCR技术进行检测,首先以Primer premier 5.0和O lig o 6.0软件设计和评价引物,对引物进行特异性检测和扩增产物测序验证,然后检测89例不同病理类型胶质瘤组织和24例非肿瘤脑组织中M AGE-D4ab和M AGE-D4c的表达,并结合胶质瘤患者临床病理参数进行统计分析.结果 所设计的引物可特异性扩增M AGE-D4ab和M AGE-D4c.所检测的组织中,M AGE-D4ab的表达普遍较M AGE-D4c丰富,其中胶质瘤组织中MAGE-D4ab的表达率明显高于非肿瘤脑组织(P<0.05),而MAGE-D4c的表达率与非肿瘤脑组织比较差异无统计学意义(P>0.05);胶质瘤组织和非肿瘤脑组织中均存在MAGE-D4ab-c和MAGE-D4ab+c两种表达形式,其中胶质瘤中MAGE-D4ab-c的表达率高于非肿瘤脑组织(P<0.05),而MAGE-D4ab+c的表达率则与非肿瘤脑组织差异无统计学意义(P>0.05);此外,MAGE-D4ab、MAGE-D4c、MAGE-D4ab-c和MAGE-D4ab+c的表达率均与性别、年龄、肿瘤直径和W HO分级等胶质瘤临床资料无显著性关联.结论 M AGE-D4ab及M AGE-D4ab-c形式在胶质瘤中异常表达,可能与胶质瘤的发生相关.

目的:研究黑色素瘤相关抗原基因-D4 (MAGE-D4)和细胞分裂周期蛋白25A(CDC25A)在人脑胶质瘤(胶质瘤)中的表达水平和临床意义,并探究二者的关系.方法:利用GEPIA在线数据库分析胶质瘤组织中MAGE-D4和CDC25A的表达情况,然后收集47例胶质瘤组织,逆转录实时荧光定量聚合酶链反应(PT-qPCR)验证MAGE-D4和CDC25A的表达,并进一步统计分析其与临床病理指标的关系及二者表达相关性.结果:GEPIA数据库显示MAGE-D4与CDC25A在胶质瘤组织中均高表达(P＜0.05);PT-qPCR结果显示所检测的胶质瘤组织MAGE-D4 mRNA和CDC25A mRNA的表达水平均明显高于正常脑组织,且二者表达呈明显正相关关系(r=0.3174,P＜0.05);此外,MAGE-D4 mRNA的表达与WHO分级和Ki-67表达相关,而CDC25A mRNA表达仅与WHO分级相关(P＜0.05).结论:CDC25A与MAGE-D4在胶质瘤组织中呈正相关关系,提示二者可能共同在胶质瘤的发生发展中发挥重要作用,可能成为胶质瘤生物治疗的潜在靶点.

Cancer testis antigens (CTAs) are attractive targets for tumor immunotherapy because of their tumor-specific expression. Since more than half of confirmed CTAs are located on the X-chromosome, we asked whether there is a link between CTA expression and X-chromosomes. Recent reports have shown that reactivation of the inactive X-chromosome, known as X-chromosome reactivation (XCR), a unique phenomenon that exists in many high-risk tumors in women, can transform the expression of many X-linked genes from monoallelic to biallelic. In this review, we discuss the link between CTA and XCR with the hopes of providing some novel insights into tumor biology.

Melanoma-associated antigen D4 (MAGE-D4) is a novel member of MAGE family. This study aimed to examine the expression and immunogenicity of MAGE-D4 in colorectal cancer (CRC) to determine its potential as a prognosis and immunotherapeutic target. The expression of MAGE-D4 mRNA and protein was determined by RT-PCR and immunohistochemistry (IHC) in CRCs with paired adjacent non-tumor tissues, colorectal adenomas and normal colorectal tissues, respectively. Sera from 64 CRC patients were tested for MAGE-D4 antibody by ELISA. MAGE-D4 mRNA was more frequently expressed in CRCs (76.7%, 46/60) than in adjacent non-tumor tissues (15.0%, 9/60). MAGE-D4 protein was detected in all the CRC tissues tested, 70.0% of which showed high expression. There was no MAGE-D4 protein detected in any paired adjacent non-tumor tissue. No MAGE-D4 expression was found in colorectal adenomas and normal colorectal tissues by either RT-PCR or immunohistochemistry. Patients with high MAGE-D4 protein expression had significantly shorter overall survival than those with low MAGE-D4 protein expression (median, 68.6 vs 122.2 months; P=0.030). Furthermore, multivariate analysis exhibited high MAGE-D4 protein expression had a trend toward an independent prognostic factor (hazard ratio: 6.124; P=0.050). Humoral immunity to MAGE-D4 was detected in 12 of 64 (18.8%) CRC patients' sera but not in 77 healthy donors. There was no correlation between MAGE-D4 expression, serum antibody and clinicopathological parameters. These findings suggest MAGE-D4 may serve as a potentially prognostic biomarker and an attractive target of immunotherapy in CRC.

Melanoma-associated antigen (MAGE) family genes have been considered as potentially promising targets for anticancer immunotherapy. MAGED4 was originally identified as a glioma-specific antigen. Current knowledge about MAGED4 expression in glioma is only based on mRNA analysis and MAGED4 protein expression has not been elucidated. In the present study, we investigated this point and found that MAGED4 mRNA and protein were absent or very lowly expressed in various normal tissues and glioma cell line SHG44, but overexpressed in glioma cell lines A172,U251,U87-MG as well as glioma tissues, with significant heterogeneity. Furthermore, MAGED4 protein expression was positively correlated with the glioma type and grade. We also found that the expression of MAGED4 inversely correlated with the overall methylation status of the MAGED4 promoter CpG island. Furthermore, when SHG44 and A172 with higher methylation were treated with the DNA demethylating agent 5-aza-2'-deoxycytidine (5-AZA-CdR) reactivation of MAGED4 mRNA was mediated by significant demethylation in SHG44 instead of A172. However, 5-AZA-CdR treatment had no effect on MAGED4 protein in both SHG44 and A172 cells. In conclusion, MAGED4 is frequently and highly expressed in glioma and is partly regulated by DNA methylation. The results suggest that MAGED4 might be a promising target for glioma immunotherapy combined with 5-AZA-CdR to enhance its expression and eliminate intratumor heterogeneity.

Targeting delivery of anticancer agents is a promising field in anticancer therapy. Inherent tumor-tropic and migratory properties of mesenchymal stem cells (MSCs) make them potential vehicles for targeting drug delivery systems for tumors. Although, MSCs have been successfully studied and discussed as a vehicle for cancer gene therapy, they have not yet been studied adequately as a potential vehicle for traditional chemical anticancer drugs. In this study, we have engineered MSCs as a potential targeting delivery vehicle for paclitaxel (TAX)-loaded nanoparticles (NPs). The size, surface charge, starving time of MSCs, incubating time and concentration of NPs could influence the efficiency of NPs uptake. In vitro release of TAX from CTS (chitosan)-TAX-NP-MSCs and the expression of P-glycoprotein demonstrated that release of TAX from MSCs might involve both passive diffusion and active transport. In vitro migration assays indicated that MSCs at passage number 3 have the highest migrating ability. Although, the migration ability of CTS-TAX-NP-MSCs could be inhibited by uptake of CTS-TAX-NPs, this ability could recover 6 days after the internalization.

Objective: To prepare and characterize the iloperidone tablets.Methods: Hydroxypropyl methyl cellulose(HPMC) was used as carrier of the solid dispersion,and lactose monohydrate and microcrystalline cellulose(MCC) as loading agents.The formulation was optimized via single factor tests.The differential scanning calorimeter(DSC) and powder X-ray diffractometry were used to identify the states of the drug existence in the product.Results: The optimized formulation was composed of 3.243% iloperidone,6.486% HPMC,11% crospovidone(PVPP,exterior addition 8% and interior addition 3%),52.314% lactose monohydrate,26.157% MCC,0.5% magnesium stearate(MS),and 0.3% colloidal silicon dioxide.Iloperidone existed in the product at amorphous forms or solvates according to DSC and X-ray diffractometry.Conclusion: The drug dissolution behavior of obtained iloperidone tablets is similar to commercial products.