BackgroundCirc0001361 is a novel circRNA identified by our previous high-throughput sequencing of glioma. Here we explored the functional involvement of circ0001361 in gliomagenesis and elucidated its potential molecular mechanisms.MethodsThe expression of circ0001361 in glioma was determined by qRT-PCR. CCK-8, colony formation assay, wound-healing, transwell assay, flow cytometric analysis and western blot were conducted to investigate cell proliferation, migration, invasion and apoptosis. The potential target miRNAs of circ0001361 and their downstream mRNAs were predicted by bioinformatics analysis and validated using dual-luciferase reporter assays.ResultsElevated expression of circ0001361 were observed in glioma tissues. The expression of circ0001361 was positively correlated with WHO tumor grades and Ki67 index, a well-established proliferation biomarker. Functional assays demonstrated that circ0001361 depletion inhibited cell proliferation, migration and invasion, while it promoted apoptosis. The bioinformatics analysis revealed that circ0001361 might target hsa-miR-525-5p, and further indicated that MEIS1 was a predicted downstream target of hsa-miR-525-5p. Subsequently, these predicted targeting relationships were both confirmed by dual-luciferase reporter assays.ConclusionCirc0001361 enhances tumorigenic properties. Mechanistically, circ0001361 may regulate glioma progression via hsa-miR-525-5p/MEIS1, suggesting its potential as a therapeutic target for glioma intervention strategies.

Glioma is a highly lethal tumor of the central nervous system (CNS) with limited therapeutic options. Recent evidence has highlighted the role of dysregulated alternative splicing in glioma progression. Although OY-TES-1 has been proposed as a potential therapeutic target, its splice isoforms have not been fully characterized. This study aimed to identify the clinically relevant splice variant of OY-TES-1 associated with glioma progression and to evaluate its potential as a target for innovative therapeutic strategies against this challenging disease. The potential splicing patterns of OY-TES-1, along with their relative frequency and correlation with patient survival, were analyzed via the TCGA SpliceSeq and OncoSplicing databases. RNA-Seq by expectation maximization (RSEM) values and clinicopathological data for all OY-TES-1 gene transcripts were downloaded from the UCSC Xena database, and Cox regression analysis was performed for both univariate and multivariate prognostic assessments. The expression of OY-TES-1 mRNA in glioma and normal brain tissues was detected via RT-PCR. The relationships between OY-TES-1 mRNA expression and the clinicopathological characteristics of glioma patients were analyzed via the χ2 test. OY-TES-1-V5a was overexpressed in glioma cells through transient transfection with plasmids as well as stable transfection with lentivirus for further functional analysis. Glioma cell proliferation was assessed via the Cell Counting Kit-8 (CCK-8) assay. Migration and invasion abilities were evaluated via wound healing, Transwell, and Transwell Matrigel assays. Apoptosis was analyzed by flow cytometry. Bioinformatic analysis revealed four alternative splice variants of OY-TES-1 in glioma, among which OY-TES-1-V5a presented a relatively high percent spliced-in (PSI) value that was associated with significantly shorter overall survival. OY-TES-1-V5a was further identified as an independent prognostic risk factor for glioma patients, as its mRNA expression was significantly associated with Karnofsky performance status (KPS), tumor grade, and isocitrate dehydrogenase 1 (IDH1) mutation status. RT-PCR validation confirmed that OY-TES-1-V5a was overexpressed in glioma tissues compared with normal brain tissues. Functionally, forced expression of OY-TES-1-V5a enhanced glioma cell proliferation, migration, and invasion while suppressing apoptosis. The OY-TES-1 splice variant V5a is highly expressed in glioma, is associated with poor prognosis, and actively drives malignant behavior, indicating its potential utility as a prognostic biomarker and a candidate target for therapeutic intervention.

While our prior study identified the HLA-A *0201-restricted ACRBP epitope peptide and demonstrated its capacity to generate cytotoxic T lymphocytes (CTLs) in vitro, the clinical relevance of the peptide-induced T cell reactivity in ovarian cancer (OC) patients and the in vivo anti-tumor efficacy of these CTLs remain unexplored. In this study, dendritic cells were sensitized with ACRBP peptide (ALLVLCYSI) and co-cultured with autologous CD8+T cells to induce the production of specific cytotoxic T lymphocytes (Pep-CTLs). The anti-tumor effects of Pep-CTLs were evaluated in SCID mice bearing human ovarian cancer (OC) OVCAR-3 cells. Concurrently, we co-cultured ALLVLCYSI peptide with peripheral blood mononuclear cells (PBMCs) from OC patients (HLA-A2+, ACRBP+) and assessed the number of specific T cells using ELISPOT assays. The immunological impact of the ACRBP peptide against human OC was validated through both in vitro and in vivo experiments. These findings establish a preclinical foundationfor developing ACRBP peptide-based vaccines in OC immunotherapy. To further elucidate ACRBP’s role in OC treatment, the study analyzed single-cell RNA sequencing data from 8 OC patients and bulk RNA sequencing data from the Cancer Genome Atlas Project (TCGA) comprising 308 ovarian cancer cases. This analysis aimed to explore the heterogeneity among ACRBP-expressing tumor cell populations and to investigate the correlation between ACRBP expression and immune molecule expression (including MHC and chemokines) alongside chemotherapy response. These insights furnish a theoretical framework supporting the future application of ACRBP in tumor immunotherapy and strategies to prevent immune escape.

Intratumoral heterogeneity poses significant challenges to the efficacy of cancer immunotherapy. Melanoma-associated antigen D4 (MAGED4) has been proposed as a potential immunotherapeutic target in oral squamous cell carcinoma (OSCC). This study aims to investigate the expression of MAGED4, focusing on its intratumoral expression heterogeneity and the underlying epigenetic regulation mechanism. Utilizing public online databases, immunohistochemical analyses of clinical specimens, and single-cell RNA sequencing data, we found that MAGED4 was overexpressed with significant intratumoral heterogeneity in OSCC tissues. Methylation-promoter luciferase reporter assays revealed that MAGED4 transcription was suppressed by DNA methylation at its promoter region. Additionally, co-expression analysis implicated a potential role for histone acetylation in regulating MAGED4. To functionally validate these findings, we treated OSCC cells with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (DAC) and histone deacetylase inhibitors trichostatin A (TSA) and valproic acid (VPA). The triple-drug combination treatment resulted in the most robust reactivation of MAGED4 expression, correlating with promoter DNA demethylation and enhanced acetylation of H3K9 and H3K27 at the MAGED4 promoter. Our findings elucidate critical epigenetic mechanisms contributing to MAGED4 heterogeneity in OSCC and highlight the potential of combination epigenetic therapies to reverse this heterogeneity, thereby providing a foundation for exploring such approaches to improve immunotherapeutic outcomes.

PURPOSE:Mitochondrial dysfunction leading to impaired energy metabolism has been recognized as a pivotal factor contributing to renal tubular epithelial cells (RTECs) damage in the context of dyslipidemia conditions in diabetic kidney disease (DKD). The primary objective of this study is to elucidate the role and underlying mechanism of the proto-oncogene Metadherin (MTDH) in mediating mitochondrial damage within this specific pathological context in vitro. METHODS:The expression of MTDH in RTECs was modulated by transfecting small interfering RNA and plasmid, while palmitic acid (PA) was employed to simulate diabetic lipid metabolism disorder. Mitochondrial damage was evaluated by examining various parameters including mitochondrial morphology, membrane potential, reactive oxygen species (ROS) production, adenosine triphosphate (ATP) production, as well as morphological and structural alterations. Additionally, Carnitine acetyltransferase (CrAT) expression was assessed using Western blotting and quantitative real-time polymerase chain reaction, and CrAT activity was quantified. RESULT:MTDH expression was upregulated in PA-induced RTECs, while CrAT expression and activity were inhibited. Downregulation of MTDH mitigated PA-induced mitochondrial damage, as demonstrated by the preservation of mitochondrial membrane potential, reduction in mitochondrial ROS production, prevention of ATP depletion, and maintenance of mitochondrial structure. This was accompanied by an upregulation in CrAT expression and activity. Conversely, overexpression of MTDH exacerbated mitochondrial dysfunction by impairing membrane potential, augmenting mitochondrial ROS production, inhibiting ATP synthesis, and suppressing CrAT expression and activity. CONCLUSION:In the context of dyslipidemia conditions, MTDH is upregulated and suppresses the expression and activity of CrAT in RTECs, thereby inducing mitochondrial dysfunction and perturbing energy metabolism. These alterations exacerbate the injury to RTECs, consequently promoting the progression of DKD.

Aim: To investigate the impact of human herpes virus (HHV) carriage on lung microbiota, and its correlation with clinical features and laboratory indicators in patients.Methods: Retrospective analysis was conducted on 30 outpatient lung infection cases, which were divided into HHV (n = 15) and non-HHV (n = 15) groups. mNGS detected microbial composition. Microbial diversity and abundance were tested using Shannon and Chao1 indices. Their relationship with laboratory indicators were explored.Results: Significant differences in microbial abundance and distribution were found between two groups (p < 0.05). Moreover, HHV group showed negative correlations (p < 0.05) between Prevotella, Porphyromonas, Streptococcus and basophil/eosinophil percentages.Conclusion: HHV carriage impacts lung microbiota, emphasizing the need for clinicians to pay attention to HHV reactivation in outpatient lung infection patients.

Cancer-testis antigen CT23 is a class of tumor-associated antigens (TAA) characterized by restricted expression in male germ cells and a variety of tumor tissues. Numerous studies have shown that CT23 is closely related to tumor cell viability, proliferation, metastasis and invasion. CT23 is immunogenic and can cause specific immune response in tumor patients. Therefore, it is considered to be one of the best target antigens for designing therapeutic tumor vaccines and T-cell-mediated tumor immunotherapy. In this study, we initially obtained seven HLA-A*0201-restricted CT23 epitope candidate peptides through the T cell epitope prediction program. Subsequently, a T2 cell binding assay revealed the potential binding of all candidate peptides with HLA-A2 molecules. Notably, peptide P7 (ALLVLCYSI) exhibited the highest affinity, as evidenced by a fluorescence index (FI) of 2.19. Dendritic cells (DCs) loaded with CT23 candidate peptide can stimulate CD8+T cell activation and proliferation, and compared with other candidate peptides, candidate peptide P7 is superior. The cytotoxic T lymphocytes (CTLs) stimulated by the peptide P7 had killing effect on tumor cells (HLA-A*0201+, CT23+), but no killing effect on tumor cells (HLA-A*0201-, CT23+). The CTLs induced by the peptide P7 also had a specific killing effect on T2 cells bearing the peptide P7. In summary, our findings suggest that the CT23 peptide P7 (ALLVLCYSI) can induce immune responses and holds potential for tumor-specific CTL therapy.

目的 探讨长链非编码RNA LINC01152在胶质瘤中的表达及其对胶质瘤细胞恶性生物学行为的影响.方法 应用LncSpA V2.0和GEPIA数据库分析LINC01152在胶质瘤中的表达;qRT-PCR检测10例正常脑组织、40例胶质瘤组织和5株胶质瘤细胞系中LINC01152 mRNA的表达.使用DAVID数据库对LINC01152共表达基因进行GO和KEGG富集分析,预测其功能.通过AnnoLnc2、TargetScan、LinkedOmics和miRDB数据库预测并筛选LINC01152相关miRNA和靶基因,构建ceRNA调控网络.利用小干扰RNA在胶质瘤细胞系下调LINC01152表达,通过CCK-8法测试、划痕愈合实验、Transwell迁移和侵袭实验、流式细胞术、Western blot等检测LINC01152对胶质瘤细胞的增殖、迁移、侵袭和凋亡等功能的影响.结果 数据库分析显示,与其他肿瘤类型相比,LINC01152在胶质母细胞瘤(GBM)和低级别胶质瘤(LGG)中高表达,且高于正常脑组织.qRT-PCR结果显示LINC01152 mRNA在胶质瘤组织中的表达显著高于正常脑组织(P<0.01).LINC01152的表达与Ki-67相关(P<0.05),但与胶质瘤患者的性别、年龄、肿瘤大小、P53蛋白、胶质纤维酸性蛋白(GFAP)、6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)和WHO分级等临床参数无关.共表达基因的功能富集分析表明,LINC01152主要参与细胞黏附和突触信号传导等生物学过程.根据预测的靶基因构建LINC01152-miRNA-mRNA调控网络.LINC01152表达下调后,A172和U87细胞的增殖、迁移、侵袭能力降低(P<0.01),而凋亡细胞数量增加(P<0.001).结论 LINC01152在胶质瘤中高表达,通过促进胶质瘤细胞的增殖、迁移、侵袭和抑制凋亡等途径促进胶质瘤的恶性生物学行为.

Objective: To screen and validate cancer testis antigens (CTAs) as potential biomarkers and explore their molecular mechanisms in glioblastoma (GBM). Methods: Ribonucleic acid sequencing (RNA-seq) and bioinformatics analyses were utilized to screen the highly expressed CTAs in GBM. Correlation analysis was used to identify poten-tial biomarkers associated with tumor purity and prognosis. Immunohistochemistry was applied for detection of pro-tein expression. Protein-protein interaction (PPI) network construction, functional enrichment analysis, and binding domain prediction were performed to investigate the underlying molecular mechanisms of GBM. Results: A total of 8 highly expressed CTAs were identified in GBM. One of them was PDZ-binding kinase (PBK). PBK messenger RNA (mRNA) was most highly expressed in GBM and associated with tumor purity and prognosis, PBK protein expression was also significantly increased in GBM tissues and correlated with p53 expression. Functional enrichment analysis revealed that the PBK related genes were predominantly enriched in cell cycle pathway with 38 genes enriched. The proteins encoding by these 38 genes were performed by binding domain prediction analysis, which demonstrated 15 proteins interacting with PBK. Most of these proteins were up regulated in GBM. Conclusion: PBK is highly ex-pressed in GBM. It may serve as a potential biomarker for GBM targeting therapy and the cell cycle modulator by interacting with certain key molecules of cell cycle in GBM.

目的 探讨人脑胶质瘤中黑色素瘤相关抗原(MAGE)-D4和钙联蛋白(CANX)mRNA的表达水平,分析其相关性及临床意义.方法 应用基因表达谱交互式分析(GEPIA)数据库数据[胶质母细胞瘤(GBM)组织163例,低级别胶质瘤(LGG)组织518例,正常脑组织207例]分析MAGE-D4和CANX mRNA在人脑胶质瘤组织中的表达情况,并通过逆转录-定量聚合酶链反应(RT-qPCR)法在临床样本(胶质瘤62例,正常脑组织11例)中进一步验证.分析MAGE-D4和CANX mRNA表达水平与胶质瘤患者临床指标的关联性.应用小干扰RNA(siRNA)技术沉默胶质瘤细胞株SF126和U251中MAGE-D4的表达,探讨MAGE-D4与CANX表达的相关性.结果 GEPIA数据库数据分析结果显示,GBM和LGG组织的MAGE-D4和CANX 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,胶质瘤组织中MAGE-D4和CANX mRNA的表达水平显著高于正常脑组织(P<0.05).MAGE-D4及CANX 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)的表达水平均无关联性(P>0.05).Pearson相关分析结果显示,临床胶质瘤样本组织中MAGE-D4与CANX mRNA表达水平呈正相关(r=0.916,P=0.000).细胞实验结果显示,经MAGE-D4 siRNA沉默后,SF126和U251细胞CANX mRNA的表达水平显著下降(P<0.05).结论 人脑胶质瘤中MAGE-D4、CANX高表达,且二者表达水平呈正相关,提示其可能共同参与了胶质瘤的发生、发展,具有成为胶质瘤免疫治疗靶点的应用前景.

Objective Glioblastoma (GBM) is the most common, invasive, and malignant primary brain tumor with a poor prognosis and high recurrence rate. It’s known that some microRNAs (miRNAs) which are associated with tumorigenesis and progression can be considered as prognostic and therapeutic targets in tumors including GBM. This study aims to highlight the potential role of the core miRNAs in GBM and their potential use as a prognostic and therapeutic biomarker. Methods Differentially expressed miRNAs (DEmiRNAs) were identified in GBM by integrating miRNA-sequencing results and a GBM microarray dataset from the Gene Expression Omnibus (GEO) database through bioinformatics tools. The dysregulated miRNAs were identified by survival analysis through Chinese Glioma Genome Atlas (CGGA). Target genes of the dysregulated miRNAs were predicted on MiRWalk and miRTarBase database. TAM2.0 database, Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis were used to analyze the function of the dysregulated miRNAs. Subsequently, protein-protein interaction (PPI) network analysis was used to identify the top 20 hub targets of the up-regulated and down-regulated miRNAs, respectively. Then, core miRNAs in GBM were identified by constructing dysregulated miRNA-differentially expressed hub gene networks. Validation of the core miRNAs expression was detected in 41 GBM tissues compared to 8 normal brain tissues. Furthermore, the potential biomarkers were identified by clinical correlation analysis and survival analysis. Results Totally, 68 intersecting DEmiRNAs were identified, 40 of which were upregulated and the other 28 miRNAs were downregulated. Two upregulated and 4 downregulated miRNAs showed prognostic significance. Most differentially expressed hub genes were regulated by the miR-28-5p and miR-1224-5p, which were respectively upregulated and downregulated in GBM. The correlation between miR-1224-5p level and recurrence was statistically significant ( P =0.011). Survival analysis showed that high miR-28-5p level and high miR-1224-5p level were both associated with better prognosis. Moreover, high miR-1224-5p level was an independent prognosis factor for GBM patients according to the cox regression analysis. Conclusion MiRNA-1224-5p could be a potential target for the prognosis and treatment in GBM.

OBJECTIVE:Glioblastoma multiforme (GBM), the most malignant intracranial neoplasm, is associated with a high mortality and recurrence rate due to the aggressive nature and heterogeneity of the tumor. Some of the molecular markers involved in the tumorigenesis of GBM are essential in prognosis, diagnosis, and treatment. Due to the limitations of therapeutic effects, this study aims to explore novel biomarkers with prognostic value and to provide new insights into therapeutic targets.METHODS:The expression profile of mRNAs in GBM was detected by RNA-sequencing, and differentially expressed genes were identified by integrating the data from RNA-seq results and the GEPIA2 database. Of the total 40 hub genes, FN1, P4HB, and PPIB showed prognostic significance based on both GEPIA2 and CGGA databases. The validation of FN1, P4HB, and PPIB expression by qPCR and correlation analysis with clinicopathological features were performed in 41 GBM tissues from our institution.RESULTS:Kaplan-Meier analysis revealed that FN1 and P4HB expressions levels were related to the overall survival (OS) of GBM patients (P<0.05). Multivariate analysis showed that FN1 overexpression (HR=9.199, P=0.002) was an independent and unfavorable prognostic factor for GBM patients. The median survival time was 8.5 months and 21 months for high and low expressions of FN1, respectively.CONCLUSION:It was suggested that FN1 could be an ideal target for prognosis and a potential therapeutic target in GBM.

目的 探讨hsa_circ_0021350在胶质瘤中的表达及临床意义,初步评价其应用于胶质瘤诊断和治疗的潜力.方法 选择2016年1月至2022年5月在广西医科大学第一附属医院神经外科经手术获取的胶质瘤组织标本40例(高级别胶质瘤22例,低级别胶质瘤18例),正常脑组织标本10例.通过RNase R消化联合逆转录聚合酶链反应(RT?PCR)验证hsa_circ_0021350的环状特性.通过TA克隆测序法鉴定实时荧光定量PCR(qRT?PCR)扩增产物.比较高级别胶质瘤、低级别胶质瘤和正常脑组织hsa_circ_0021350的表达水平.分析hsa_circ_0021350表达与胶质瘤患者临床病理指标的关联性.结果 该研究所设计的发散引物可有效特异扩增hsa_circ_0021350.经验证,hsa_circ_0021350呈环状,且能耐受RNase R消化.qRT?PCR实验结果显示,高级别胶质瘤和低级别胶质瘤hsa_circ_0021350的表达水平均高于正常脑组织,且高级别胶质瘤hsa_circ_0021350的表达水平较低级别胶质瘤更高,差异有统计学意义(P<0.05).hsa_circ_0021350的表达与胶质瘤分级和ki67阳性率具有显著关联性(P<0.05).结论 hsa_circ_0021350在胶质瘤,尤其是高级别胶质瘤中呈高表达,且与肿瘤增殖因子ki67相关,是胶质瘤诊断和治疗的潜在分子靶点.

Background A glioma is a tumor originating from glial cells in the central nervous system. Although significant progress has been made in diagnosis and treatment, most high-grade glioma patients are prone to recurrence. Therefore, molecular targeted therapy may become a new direction for adjuvant therapy in glioma. In recent years, many studies have revealed that circular RNA (circRNA) may play an important role in the occurrence and development of many tumors including gliomas. Our previous study found that the expression of hsa_circ_0008922 was up-regulated in glioma tissues upon RNA sequencing. The biological mechanism of circ_0008922 is still unreported in gliomas. Therefore, in this study, we preliminarily outlined the expression of hsa_circ_0008922 in glioma and explored its biological functions. Methods The expression of hsa_circ_0008922 in forty glioma tissues and four glioma cell lines (A172, U251, SF763 and U87) was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The correlation between hsa_circ_0008922 expression and clinicopathological features of glioma patients was evaluated by Fisher’s exact test. To understand the potential function of hsa_circ_0008922 in glioma, we constructed small interfering RNA (siRNA) to hsa_circ_0008922 to downregulate its expression in glioma cell lines A172 and U251. With these hsa_circ_0008922 downregulated cells, a series of assays were carried out as follows. Cell proliferation was detected by CCK8 assay, migration and invasion were determined by wound healing assay and transwell assay, respectively. Colony formation ability was evaluated by plate clonogenic assay. Moreover, flow cytometry combined with Western blot was performed to analyze apoptosis status and the expression of apoptotic related proteins (caspase 3 and caspase 9). Finally, the possible biological pathways and potential miRNA targets of hsa_circ_0008922 were predicted by bioinformatics. Results We found that the expression of hsa_circ_0008922 in glioma tissues was 3.4 times higher than that in normal tissues. The expression of has_circ_0008922 was correlated with WHO tumor grade. After down-regulating the expression of hsa_circ_0008922, malignant biological behavior of glioma cells was inhibited, such as cell proliferation, colony formation, migration, and invasion. At the same time, it also induced apoptosis of glioma cells. Predicted analysis by bioinformatics demonstrated that hsa_circ_0008922 may be involved in tumor-related pathways by acting as a molecular sponge for multiple miRNAs (hsa-let-7e-5p, hsa-miR-506-5p, hsa-let-7b-5p, hsa-let-7c-5p and hsa-let-7a-5p). Finally, we integrated our observation to build a circRNA-miRNA-mRNA predictive network.

Objective The study evaluated the efficacy of combined epigenetic drugs of decitabine (DAC), valproic acid (VPA), and trichostatin A (TSA) on immunotherapy against glioma. Methods The expression and prognosis of MAGE-D4 in glioma were analyzed online, and the expression of MAGE-D4 and HLA-A2 in glioma induced by epigenetic drugs was detected by qRT-PCR, Western blot, and flow cytometry. The methylation status of the MAGE-D4 promoter was determined by pyrosequencing. An HLA-A2 restricted MAGE-D4 peptide was predicted and synthesized. An affinity assay and a peptide/HLA complex stability assay were performed to determine the affinity between peptide and HLA. CCK8 assay, CFSE assay, ELISA and ELISPOT were performed to detect the function of MAGE-D4 peptide-specific T cells. Flow cytometry, ELISA, and cytotoxicity assays were used to detect the cytotoxicity effect of MAGE-D4 peptide-specific T cells combined with epigenetic drugs against glioma in vitro. Finally, the glioma-loaded mouse model was applied to test the inhibitory effect of specific T cells on gliomas in vivo. Results MAGE-D4 was highly expressed in glioma and correlated with poor prognosis. Glioma cells could be induced to express MAGE-D4 and HLA-A2 by epigenetic drugs. MAGE-D4-associated peptides were found that induce DCs to stimulate the highest T-cell activities of proliferation, IL-2 excretion, and IFN-γ secretion. MAGE-D4 peptide-specific T cells treated with TSA only or combining TSA and DAC had the most cytotoxicity effect, and its cytotoxicity effect on glioma cells decreased significantly after HLA blocking. In vivo experiments also confirmed that MAGE-D4-specific T cells inhibit TSA-treated glioma. Conclusion MAGE-D4 is highly expressed in glioma and correlated with the prognosis of glioma. The novel MAGE-D4 peptide identified was capable of inducing MAGE-D4-specific T cells that can effectively inhibit glioma growth, and the epigenetic drug application can enhance this inhibition.

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 构建肾嫌色细胞癌(CRCC)的内源竞争性RNA(ceRNA)调控网络,筛选出与CRCC预后相关的信使RNA(mRNA)、长非编码RNA(lncRNA)和微小RNA(miRNA),为CRCC的诊断和预后提供理论依据.方法 提取TCGA数据库中CRCC样本(65例)和正常肾样本(24例)的转录组数据,以错误发现率(FDR)<0.05,差异倍数的对数绝对值(log2|FC|)>2为条件筛选差异表达(DE)的lncRNA、miRNA和mRNA,据此构建ceRNA调控网络.应用Kaplan-Meier法探讨与CRCC患者预后相关的因子,据此进一步构建蛋白-蛋白互作(PPI)网络,并进行GO与KEGG富集分析.结果 共筛选出3679个DElncRNA,297个DEmiRNA和5914个DEmRNA.mRNA下调的ceRNA网络包含DEmiRNA 95个、DEmRNA 268个、DElncRNA 737个;mRNA上调的ceRNA网络包含DEmiRNA 85个、DEmRNA 234个、DElncRNA 924个.Kaplan-Meier分析筛选出3个DEmRNA(DEPDC1、SLC7A11和E2F7)和1个DEmiRNA(miR-26b-5p)与CRCC预后存在关联(P<0.05).以DEPDC1、SLC7A11和E2F7构建的PPI网络中共含有8个mRNA(TOP2A、DEPDC1、SLC3A2、E2F8、SLC7A11、E2F1、TP53和E2F7)参与蛋白互作,GO和KEGG富集分析显示,这些互作蛋白功能主要富集于DNA损伤反应、p53介导的细胞组织与启动子特异结合、铁死亡,以及肿癌疾病相关信号通路.结论 该研究通过构建CRCC的ceRNA调控网络筛选出与其预后相关的关键RNAs,为CRCC的诊断和治疗提供了理论依据.

目的 通过生物信息学分析PDZ结合激酶/T-淋巴因子激活的杀伤细胞来源的蛋白激酶(PBK/TOPK)在乳腺癌(BRCA)中的表达及临床意义,并预测其潜在的分子通路和生物学功能.方法 挖掘Oncomine数据库分析PBK/TOPK信使核糖核酸(mRNA)在不同癌症中的表达情况.下载癌症基因组图谱(TCGA)数据库BRCA和正常乳腺组织的RNA测序数据和临床信息,分析PBK/TOPK mRNA表达与临床特征的相关性.应用Kaplan-Meier Plotter数据库进行生存分析.通过筛选PBK/TOPK相关基因,并进行功能富集分析,预测PBK/TOPK在BRCA中的潜在分子通路和生物学功能.结果 PBK/TOPK基因在BRCA组织中的表达显著高于正常乳腺组织(P<0.05),并与雌激素受体(ER)、孕激素受体(PR)、三阴性乳腺癌(TNBC)和绝经状态存在关联性(P<0.05).生存分析结果显示,PBK/TOPK基因高表达提示BRCA预后不良(P<0.05).功能富集分析显示PBK/TOPK相似基因主要涉及BRCA细胞分裂、细胞周期调节和微管细胞骨架形成等生物功能,参与BRCA细胞周期等信号通路.结论 PBK/TOPK mRNA在BRCA组织中高表达,与BRCA患者预后不良密切相关,可能是BRCA潜在的生物标志物和治疗靶标.

Abstract Introduction Cancer testis (CT) antigens are attractive targets for cancer immunotherapy because of their expression restriction and immunogenicity. The acrosin binding protein (ACRBP) is a member of CT antigens. This study aimed to evaluate ACRBP expression and immunogenicity in ovarian cancer (OC). Methods The expression level of ACRBP in OC tissues, normal ovarian tissues, and cell lines was detected via quantitative real‐time polymerase chain reaction (qRT‐PCR) and immunohistochemistry. We determined the levels of ACRBP antigen and antibody in serum samples collected from patients with OC and healthy donors using enzyme‐linked immunosorbent assays (ELISA), the level of ACRBP in cell‐cultured medium was also tested. Results ACRBP mRNA and protein expressions were upregulated in OC tissues relative to normal tissue, especially highly expressed in epithelial ovarian cancer (EOC). Moreover, ACRBP expression was significantly correlated with International Federation of Gynecology and Obstetrics (FIGO) stage and chemosensitivity. Serological analysis showed that anti‐ACRBP antibody was detected in the sera of 16 of the 56 (28.5%) patients with OC but not in healthy donors. The area under the receiver operating characteristic curve for ACRBP antibody was 0.802 (95% confidence interval [CI]: 0.708–0.876), and the sensitivity and specificity for ACRBP antibody was 85.71% and 55.0%, respectively. Kaplan–Meier analysis revealed that the overall survival (OS) and disease‐free survival (DFS) in OC patients with high ACRBP expression were significantly lower than those with low expression (p = 0.040, p = 0.021). However, ACRBP antibody level was not associated with prognosis. Conclusion ACRBP expression was upregulated in OC tissues and induced humoral immune response in patients with OC, suggesting that ACRBP is a potential prognostic biomarker and a target of tumor immunotherapy for OC.

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.