Myeloid-derived suppressor cells (MDSCs) play a pivotal role in establishing an immunosuppressive tumor microenvironment (TME), yet the mechanisms underlying their functional activation remain incompletely defined. Here, we identify the Fgl2-FcγRIIB signaling axis as a critical mediator of MDSC-driven immune evasion across solid tumors. Analysis of clinical specimens revealed that Fgl2 expression is significantly elevated in tumor tissues and inversely correlates with CD8+ T cell infiltration, while positively associating with the accumulation of FcγRIIB+ MDSCs and poor patient prognosis. We demonstrate that tumor-derived exosomes (TEX) function as efficient carriers that deliver membrane-bound Fgl2 (mFgl2) to MDSCs. These exosomes are internalized by MDSCs through FcγRIIB-mediated endocytosis, leading to an enhanced immunosuppressive function characterized by upregulated arginase-1 (Arg-1) and inducible nitric oxide synthase (iNOS) expression and an increased capacity to suppress CD8+ T cell proliferation. Genetic ablation of FcγRIIB or antibody-mediated neutralization of Fgl2 abolished this exosome-mediated immunosuppressive programming, restoring T cell activity and impairing tumor growth in vivo. Importantly, a therapeutic strategy combining an exosome secretion inhibitor, in combination with PD-L1 blockade and MDSCs depletion, synergistically achieved potent antitumor effects. Our findings unveil a novel exosome-dependent mechanism through which tumors systemically educate MDSCs, establishing the Fgl2-FcγRIIB axis as a promising broad-spectrum target for cancer immunotherapy.

Objective Inducing tumor cell senescence represents a promising strategy in cancer therapy. This study aimed to elucidate the expression profile, biological functions, and molecular regulatory mechanisms of centrosomal protein 55 (CEP55) in glioma, thereby providing novel therapeutic targets and a theoretical basis for targeted glioma treatment. Methods Human glioma cell lines U251 and U87 were divided into negative control (NC), CEP55 knockdown (siCEP55-1, siCEP55-2), and forkhead box M1 (FOXM1) overexpression groups, with three biological replicates per group. CEP55 was knocked down via siRNA transfection, and FOXM1 was overexpressed using lentiviral vectors. mRNA and protein levels of CEP55 and FOXM1 were assessed by qRT-PCR and Western blotting. The mRNA expression of senescence-associated genes, including insulin-like growth factor binding protein 3 (IGFBP3), interleukin 6 (IL6), serpin family E member 1 (SERPINE1), cell division cycle 25A (CDC25A), C-X-C motif chemokine ligand 8 (CXCL8), MYC, ZFP36 ring finger protein like 1 (ZFP36L1), and ZFP36 ring finger protein like 2 (ZFP36L2), was measured by qRT-PCR. Whole-transcriptome profiling was performed by RNA sequencing (RNA-seq). Differentially expressed genes were identified using the DESeq2 package following gene quantification, and subsequent Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted. Cell cycle distribution was analyzed by flow cytometry, mitochondrial morphology was observed via transmission electron microscopy, and reactive oxygen species (ROS) levels were quantified using the DCFH-DA fluorescent probe. Senescent cells were identified by senescence-associated u03B2-galactosidase (SA-u03B2-gal) staining. Chromatin immunoprecipitation (ChIP) assay was employed to verify the binding of FOXM1 to the CEP55 promoter. Bioinformatics analysis was performed to evaluate the correlation between CEP55 expression and prognosis using data from The Cancer Genome Atlas (TCGA). A subcutaneous xenograft tumor model was established by injecting U87 cells into nude mice. Ten 6-week-old female BALB/c nude mice (body weight 18 to 22 g) were stratified and randomly assigned to the NC group or siCEP55-1 group (n=5), receiving subcutaneous inoculation of cell suspensions into the right flank. Upon tumor formation, xenografts were excised to measure longitudinal and transverse diameters. Results CEP55 expression was significantly elevated in glioma tissues compared to normal tissues, and patients with high CEP55 expression exhibited markedly shorter overall survival than those with low expression (Pu0026lt;0.001). Transfection with siCEP55-1/2 significantly downregulated CEP55 mRNA and protein levels in U251 and U87 cells (Pu0026lt;0.001) and induced prominent G0/G1 phase arrest (Pu0026lt;0.05). RNA-seq analysis identified 524 differentially expressed genes (249 upregulated, 275 downregulated), with upregulated genes significantly enriched in cellular senescence, mitogen-activated protein kinase signaling, and transcriptional dysregulation in cancer pathways. CEP55 knockdown led to significant upregulation of senescence-related genes (IGFBP3, IL6, SERPINE1, CDC25A, CXCL8, MYC, ZFP36L1; Pu0026lt;0.01), pronounced mitochondrial fragmentation, elevated intracellular ROS levels (Pu0026lt;0.01), increased SA-u03B2-gal-positive cell proportion (Pu0026lt;0.01), and markedly suppressed xenograft growth with reduced tumor volumes (P=0.006). FOXM1 expression correlated positively with CEP55 levels (r=0.898, Pu0026lt;0.0001). FOXM1 overexpression significantly upregulated CEP55 expression (Pu0026lt;0.001), and ChIP assays confirmed direct binding of FOXM1 to the CEP55 promoter region. Conclusion CEP55 is directly transcriptionally activated by FOXM1 and promotes glioma progression by suppressing cellular senescence signaling pathways.

Lung adenocarcinoma (LUAD) has emerged as both the most frequently diagnosed malignancy and the predominant contributor to cancer-related mortality worldwide. Current clinical evidence indicates that a significant proportion of LUAD cases exhibit tumor cells characterized by accelerated proliferative activity, which contributes to the aggressive biological behavior. Six microarray data sets were retrieved from the Gene Expression Omnibus (GEO), and differentially expressed genes (DEGs) were identified using the robust rank aggregation (RRA) method. The mRNA and protein levels of selected genes were subsequently validated by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and western blot (WB). Short interfering RNA (siRNA)-mediated knockdown combined with EdU incorporation assays was employed to assess proliferation in LUAD cell lines. Chromatin immunoprecipitation (ChIP) assays confirmed that FOXM1 directly regulates the transcription of its target genes. A total of 291 DEGs (133 up-regulated and 158 down-regulated) were identified. Up-regulated genes were significantly enriched in cell-cycle pathways. The FOXM1 exhibited the strongest correlation with these cell-cycle genes and was shown by ChIP-seq to bind to the promoters of 49 of them. TOP2A , MELK , CENPF , NEK2 , and KIF20A are the top 5 genes for further analysis in the The Cancer Genome Atlas (TCGA) database. These 5 genes are all highly expressed and show a worse prognosis in LUAD. Cell experiments showed that FOXM1 knockdown only inhibited the expression of CENPF and NEK2. Knocking down either FOXM1 or CENPF can inhibit the proliferation of LUAD cells. Overexpression of FOXM1 promoted CENPF expression and the proliferation of lung cancer cells. The predicted regulatory network of FOXM1 shows significant discrepancies with experimental validation data. Therefore, FOXM1’s regulatory role in the cell cycle requires further experimental verification.

Optimizing irradiation volumes and evaluating the effect of dose escalation on total and fractionated doses are critical for improving outcomes in high-grade glioma (HGG). To assess the efficacy of modified target delineation guided by multimodal magnetic resonance imaging and white matter tracts combined with moderately hypofractionated simultaneous boost intensity-modulated radiotherapy (HSIB-IMRT) in patients with newly diagnosed HGG. This single-center, 2-arm, open-label randomized clinical trial enrolled 154 patients aged 18 to 70 years with histologically confirmed, newly diagnosed HGG at a Chinese medical center from January 1, 2018, to August 31, 2022. Follow-up was completed in June 2024. Patients were randomized to receive modified target delineation guided by multimodal magnetic resonance imaging and white matter tracts combined with HSIB-IMRT (experimental arm) or standard IMRT per guideline recommendations (standard arm). Both arms received concurrent and adjuvant temozolomide chemotherapy. The primary end point was progression-free survival (PFS). The secondary end point was overall survival (OS). Among 154 enrolled patients (76 in the experimental arm and 78 in the standard arm; 85 [55.2%] male; median [range] age, 51.5 [23.0-70.0] years), the median (range) follow-up duration was 22 (4-76) months, with 96 deaths by June 2024. The median PFS was 15.5 months (95% CI, 11.7-19.3 months) in the experimental arm and 13.5 months (95% CI, 8.7-18.3 months) in the standard arm (P = .89). The median OS was 27.0 months (95% CI, 13.9-40.1 months) in the experimental arm and 21.0 months (95% CI, 18.0-24.0 months) in the standard arm (P = .24). The clinical target volume in the experimental arm (CTV1: median [range], 116.7 [20.2–370.7 cm3]; CTV2: median [range], 174.4 [34.5-463.2 cm3]) was significantly smaller than the clinical target volume in the standard arm (median [range], 225.0 [70.2-542.1 cm3]; P < .001). Recurrence rates within, outside, and multicentric to the target volume were comparable between arms. Grade 3 or 4 adverse events occurred in 4 patients (5.3%) in the experimental arm and 3 (3.8%) in the standard arm (P = .72). In this randomized clinical trial, modified target delineation with HSIB-IMRT demonstrated comparable PFS and OS to standard IMRT in patients with newly diagnosed HGG, while significantly reducing the irradiation target volume without increasing the recurrence rates outside the target volume. These results suggest valuable insights for future research aimed at personalized, reduced volume strategies to optimize outcomes and minimize neurotoxicity in HGG. ChiCTR.org.cn Identifier: ChiCTR1800014396

Importance:Optimizing irradiation volumes and evaluating the effect of dose escalation on total and fractionated doses are critical for improving outcomes in high-grade glioma (HGG). Objective:To assess the efficacy of modified target delineation guided by multimodal magnetic resonance imaging and white matter tracts combined with moderately hypofractionated simultaneous boost intensity-modulated radiotherapy (HSIB-IMRT) in patients with newly diagnosed HGG. Design, Setting, and Participants:This single-center, 2-arm, open-label randomized clinical trial enrolled 154 patients aged 18 to 70 years with histologically confirmed, newly diagnosed HGG at a Chinese medical center from January 1, 2018, to August 31, 2022. Follow-up was completed in June 2024. Interventions:Patients were randomized to receive modified target delineation guided by multimodal magnetic resonance imaging and white matter tracts combined with HSIB-IMRT (experimental arm) or standard IMRT per guideline recommendations (standard arm). Both arms received concurrent and adjuvant temozolomide chemotherapy. Main Outcomes and Measures:The primary end point was progression-free survival (PFS). The secondary end point was overall survival (OS). Results:Among 154 enrolled patients (76 in the experimental arm and 78 in the standard arm; 85 [55.2%] male; median [range] age, 51.5 [23.0-70.0] years), the median (range) follow-up duration was 22 (4-76) months, with 96 deaths by June 2024. The median PFS was 15.5 months (95% CI, 11.7-19.3 months) in the experimental arm and 13.5 months (95% CI, 8.7-18.3 months) in the standard arm (P = .89). The median OS was 27.0 months (95% CI, 13.9-40.1 months) in the experimental arm and 21.0 months (95% CI, 18.0-24.0 months) in the standard arm (P = .24). The clinical target volume in the experimental arm (CTV1: median [range], 116.7 [20.2-370.7 cm3]; CTV2: median [range], 174.4 [34.5-463.2 cm3]) was significantly smaller than the clinical target volume in the standard arm (median [range], 225.0 [70.2-542.1 cm3]; P < .001). Recurrence rates within, outside, and multicentric to the target volume were comparable between arms. Grade 3 or 4 adverse events occurred in 4 patients (5.3%) in the experimental arm and 3 (3.8%) in the standard arm (P = .72). Conclusions and Relevance:In this randomized clinical trial, modified target delineation with HSIB-IMRT demonstrated comparable PFS and OS to standard IMRT in patients with newly diagnosed HGG, while significantly reducing the irradiation target volume without increasing the recurrence rates outside the target volume. These results suggest valuable insights for future research aimed at personalized, reduced volume strategies to optimize outcomes and minimize neurotoxicity in HGG. Trial Registration:ChiCTR.org.cn Identifier: ChiCTR1800014396.

Lung cancer is a leading cause of cancer-related mortality worldwide, with non-small cell lung cancer (NSCLC) constituting the majority, and its main subtype being lung adenocarcinoma (LUAD). Despite substantial advances in LUAD diagnosis and treatment, early diagnostic biomarkers inadequately fulfill clinical requirements. Thus, we conducted bioinformatics analysis to identify potential biomarkers and corresponding therapeutic drugs for early-stage LUAD patients. Here we identified a total of 10 differentially expressed genes (DEGs) with survival significance through the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). Subsequently, we identified a promising small molecule drug, Aminopurvalanol A, based on the 10 key genes using the L1000FWD application, which was validated by molecular docking followed by in vivo and in vitro experiments. The results highlighted TOP2A, CDH3, ASPM, CENPF, SLC2A1, and PRC1 as potential detection biomarkers for early LUAD. We confirmed the efficacy and safety of Aminopurvalanol A, providing valuable insights for the clinical management of LUAD.

Immune checkpoint inhibitors (ICIs) have been widely applicated in clinical therapy in recent years. Skin-related adverse reaction is one of the most common adverse events for ICIs. Stevens-Johnson syndrome (SJS) is one of the serious cutaneous reactions threatening the life. Here, we reported a case of 76-year-old male patient with poorly differentiated metastatic lung adenocarcinoma, after 9 weeks exposure of sintilimab (3 doses) combined with paclitaxel liposome after concurrent chemotherapy/radiotherapy, experienced Stevens-Johnson syndrome involving limbs, trunk, lip and the oral mucosa. Biopsy of the skin tissue showed infiltration of CD4 and CD8 positive T lymphocytes. We also found PD-L1 expression in the glands and the basal layer of the skin. This finding is distinct from the previously reported expression of PD-L1 on the surface of epidermal keratinocytes in patients with SJS due to immunotherapy.

Background: Ovarian cancer (OV) is a highly lethal disease, and the fifth leading cause of all cancerrelated deaths in women. The study aimed to identify potential key genes associated with the proliferation and prognosis of OV. Methods: Differentially expressed genes (DEGs) between ovarian cancer and normal tissues were screened by the robust rank aggregation (RRA) method. The expression of CENPA and MYBL2 were examined in SKOV3 and A2780 ovarian cancer cell lines and tumor tissues by qRT-PCR and western blot. Small RNA interference assays, plasmid overexpression assays and EdU assays were used to validate the proliferative effect of the MYBL2-CENPA axis in ovarian cancer cell lines. The ChIP assay was used to verify the direct regulation of MYBL2 on CENPA. Results: 133 up-regulated genes and 158 down-regulated genes were identified, and the up-regulated genes mainly enrichment in cell cycle. The three up-regulated gene with DNA separation (CENPA, CENPF and CEP55) might be tightly correlated with proliferation and prognosis of OV. Knockdown CENPA expression inhibited the proliferation of A2780 and SKOV3 cells After the knockout of MYBL2, the expression of CENPA significantly decreased. MYBL2 directly binds to the promoter region of CENPA. Conclusions: The MYBL2-CENPA pathway plays an important role in the proliferation of ovarian cancer cells, suggesting that this pathway may be a potential target for the treatment of ovarian cancer.

BACKGROUND:Primary hepatic angiosarcoma (PHA) is a rare malignant tumor. We explored the demographic features and prognostic factors of PHA.METHODS:We used the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) database to extract patients diagnosed with PHA from 1975 to 2016. We used the Kaplan-Meier method and Cox proportional hazards regression to evaluate the risk factors for overall survival (OS) and disease-specific survival (DSS). The nomograms were constructed and validated using the concordance index (C-index) and calibration plots.RESULTS:In total, 366 patients were included in this study. The disease onset was hidden, and most patients already had advanced disease when diagnosed. The prognosis of PHA was very poor, and the overall 6-month, 1-year and 2-year survival rates were 20.3%, 12.8% and 9.3%, respectively. Sex, age and surgery were all predictors of both OS and DSS in multivariate analysis. Women had better survival rates than men, and patients aged <60 years benefited from surgery in the multivariate models. The nomograms presented good accuracy, with C-index values of 0.679 and 0.665 for the OS and DSS prognostic models, respectively. The calibration plots showed good agreement between the nomogram predictions and actual observations.CONCLUSIONS:PHA has a poor prognosis. Regular physical examinations are essential for the elderly. Patients aged <60 years could benefit from surgery. We constructed accurate nomograms to predict survival that can greatly benefit clinicians.

Background and Purpose To directly reveal the change in genome mutation, RNA transcript of tumor cells, and tumor microenvironment (TME) after stereotactic body radiotherapy (SBRT) in paired human lung tumor specimens. Materials and Methods Paired tumor samples were collected from 10 patients with non-small cell lung cancer (NSCLC) or lung metastatic carcinoma within a week before and after SBRT. DNA and RNA of tumor tissues was extracted from the paired samples. Whole-exome and RNA sequencing assays were performed by next-generation sequencing. Gene mutation, genomic expression, T-cell receptor (TCR) repertoire, and profiling of tumor-infiltrating immune cells were analyzed through bioinformatics analysis in paired tumor samples. CD8+ T-cell infiltration and PD-L1 expressions were detected by immunostaining in tumor tissues. Results The diversity of TCR repertoire and PD-L1 expression increased significantly in the TME, and the most enriched term of the gene ontology analysis was the immune response gene after receiving SBRT. SBRT induced neo-mutation of genes in tumor cells but did not increase tumor mutation burden in tumor tissues. TME displayed complex immune cell changes and infiltration and expression of immune-regulating factors such as C-X-C motif chemokine (CXCL) 10, CXCL16, interferons (IFNs), and IFN receptors. CD8+ T-cells in tumor tissues did not improve significantly after SBRT while the infiltrating TH1 and TH2 cells decreased remarkably. Conclusion SBRT improved the TCR repertoire diversity and PD-L1 expression in the TME and induced neo-mutation of genes in tumor cells but did not increase CD8+ T-cell infiltration and IFN expression in the tumor tissue within a week.

Abstract There were no ideal markers to predict the development of radiation pneumonitis (RP). We want to investigate the value of variations of lymphocytes and T lymphocyte subsets in predicting RP after radiotherapy (RT) of lung cancer based on previous clinical findings. A total of 182 lung cancer patients who received RT were retrospectively analyzed. Circulating lymphocytes and T lymphocyte subsets were measured before, during, and after RT. Patients were evaluated from the start of RT to 6 months post‐RT. A mice model with acute radiation‐induced lung injury was established and circulating lymphocytes were measured weekly until 8 weeks after irradiation. Univariate and multivariate analyses were adopted to identify risk factors of RP. Lymphocyte levels significantly decreased (P < .001) in patients before RP symptoms developed that also was able to be seen in the mice model and the values recovered during remission of symptoms. The decrease in lymphocyte count reflected the severity of RP. Meanwhile, CD4+ T lymphocyte count was significantly lower during the occurrence of symptoms in patients with RP than in those without RP (P < .001), and it improved along with RP recovery. Levels of lymphocytes and CD4+ T lymphocyte subsets proved as independent predictors of RP. Here we showed that lower peripheral blood levels of lymphocytes and CD4+ T lymphocyte were associated with an increased risk of RP, which was validated by this mice model, and thus are associated with differences in radiation‐induced lung toxicity among individuals and help identify those who are susceptible to developing RP after RT.

Lipopolysaccharide-induced tumor necrosis factor alpha factor (LITAF), also called p53-induced gene 7 (PIG7), was identified as a transcription factor that activates transcription of proinflammatory cytokines in macrophages in response to lipopolysaccharide (LPS). Previous studies have identified LITAF as a potential tumor suppressor in several neoplasms, including prostate cancer, B-NHL, acute myeloid leukemia, and pancreatic cancer. However, the expression and function of LITAF in human glioma remain unexplained. The present study aimed to analyze the regulation of LITAF in gliomas. Data from The Cancer Genome Atlas (TCGA) database revealed that LITAF mRNA expression in glioma tissues was higher than that in normal brain tissues, and lower LITAF expression in gliomas showed a good prognosis in patients who received radiotherapy, by Kaplan–Meier analysis. In our collected specimens, however, LITAF showed low expression in glioma tissues compared to that in the normal brain tissue. Proliferation and apoptosis of glioma cells were not affected by knockdown or overexpression of LITAF in glioma U251, U373, and U87 cells, but LITAF was able to enhance the radiosensitivity of glioma cells. Furthermore, we found that LITAF enhanced radiosensitivity via FoxO1 and its specific downstream targets BIM, TRAIL, and FASLG. Taken together, our present results demonstrate that LITAF expression is decreased in glioma tissues and might enhance radiosensitivity of glioma cells via upregulation of the FoxO1 pathway.

目的 探讨转录因子E2F1上调细胞周期蛋白A2 (CCNA2)表达对卵巢癌细胞株SKOV3增殖的影响.方法 利用TCGA网站分析卵巢癌组织与卵巢正常组织中E2F1与CCNA2的mRNA表达情况,利用TCGA卵巢癌RNA-seq数据分析E2F1基因与CCNA2基因表达相关性,利用小RNA敲低E2F1表达,利用qPCR检测E2F1、CCNA2 mRNA表达水平,利用western blot检测E2F1、CCNA2蛋白表达水平,EDU流式检测法检测SKOV3细胞增殖情况,利用网站预测CCNA2启动子区域E2F1结合位点.结果 卵巢癌组织中E2F1及CCNA2的表达明显高于正常卵巢组织,卵巢癌中E2F1表达与CCNA2表达具有相关性,敲低E2F1表达会减少CCNA2的表达,敲低E2F1的表达会抑制SKOV3细胞增殖,CCNA2转录起始位点附近存在E2F1转录因子的结合位点.结论 E2F1上调CCNA2表达促进卵巢癌细胞系SKOV3细胞的增殖.

Objective To investigate the expression of LITAF in human glioblastoma tissues and the effect of LITAF silence on the proliferation, apoptosis and radiosensitivity of glioblastoma U251 cells. Methods The expression of LITAF in glioblastoma was analyzed in The Cancer Genome Atlas (TCGA) database. The proliferation and apoptosis of U251 cells were detected with EDU kit and flow cytometry after the expression of LITAF was silenced. Clonogenic assay and flow cytometry were performed to assess the radiosensitivity of U251 cells with LITAF RNAi. Results The expression of LITAF was increased significantly in human glioblastoma tissues, compared with normal brain tissues. The proliferation and apoptosis of U251 cells weren' t interfered in U251 cells after LITAF knockdown by RNAi. After irradiation, the apoptosis was decreased remarkably and the clone formation was increased markedly in U251 cells with LITAF RNAi, compared with the control cells. Conclusion LITAF is highly expressed in human glioblastoma tissues. Knockdown of LITAF expression has no effect on the proliferation or apoptosis but decreases the radiosensitivity of U251 cells.

Abstract Complement aids in the construction of an immunosuppressive tumor microenvironment. Tumor cell–derived C3 has been previously reported, but whether and how it acts on antitumor immunity remains to be elucidated. Here, we describe a mechanism for tumor cell–derived C3 in suppressing antitumor immunity. Tumor cell–derived C3 was activated intracellularly, which results in generation of C3a. C3a modulated tumor-associated macrophages via C3a-C3aR-PI3Kγ signaling, thereby repressing antitumor immunity. Deletion of C3 in tumor cells that had high C3 expression enhanced efficacy of anti–PD-L1 treatment. Collectively, our results suggest tumor cell–derived C3 may be a useful target for cancer immunotherapy and that targeting C3 in tumor cells may enhance antitumor immunity.

Ovarian tumor domain-containing ubiquitin aldehyde binding protein 1 (OTUB1) is overexpressed in many cancers and plays an important role in tumor progression and metastasis. However, the molecular mechanisms underlying OTUB1 overexpression are not clear. In this study, we found that estrogen-related receptor alpha (ERRα, also called NR3B1) binds to OTUB1 promoter and regulates its expression in colorectal cancer. Furthermore, ERRα promoted the migration of CRC cells by inducing vimentin expression via OTUB1. Our data show that OTUB1 is a novel target of ERRα and indicate that ERRα-OTUB1 signaling may play a significant role in CRC metastasis.

Purpose Centromere protein U (CENPU) abnormally exhibits high expression in various types of human tumor tissues and participates in tumor progression; however, its expression pattern and biological function in lung cancer have not yet been elucidated. In the present study, we explored the clinical significance and biological function of CENPU in lung cancer. Materials and methods The Cancer Genome Atlas (TCGA) data analyses, quantitative real-time PCR (RT-PCR), and Western blotting were performed to quantify CENPU and FOXM1 expression in non-small-cell lung cancer (NSCLC) samples. Survival data were obtained from Kaplan-Meier plotter or PROGgene V2 prognostic database. The function of CENPU in lung cancer cell proliferation was determined using 5-ethynyl-2'-deoxyuridine (EdU), Cell Counting Kit-8 (CCK-8), and cell cycle assays, and the underlying mechanism was determined through bioinformatic analyses and validated by in vitro siRNA or plasmid transfection experiments. Results CENPU was abnormally overexpressed in NSCLC samples compared with matched paired normal tissues. Higher expression of CENPU predicted worse overall survival (OS) and relapse-free survival (RFS) in NSCLC patients. Knockdown of CENPU expression by siRNA significantly inhibited proliferation and delayed cell cycle progression of lung cancer cells. To figure out the mechanism, bioinformatic analyses were performed and the results showed that the transcription factor, FOXM1, positively correlated with CENPU. Further in vitro experiments indicated that FOXM1 was the possible downstream transcription factor of CENPU as the knockdown of CENPU led to lower expression of FOXM1 and the overexpression of FOXM1 significantly reversed the inhibition of proliferation caused by CENPU knockdown. Furthermore, FOXM1 was highly expressed in NSCLC. The knockdown of FOXM1 also attenuated proliferation and induced G1 arrest in lung cancer cells. Conclusion CENPU was highly expressed in NSCLC tissues, wherein it promoted lung cancer cell proliferation via the transcription factor, FOXM1, which could be a potential target for therapeutic strategies.

The epidermal growth factor receptor (EGFR) is often amplified in glioma, with the most common extracellular domain mutation being EGFR variant III (EGFRvIII). Abnormal EGFRvIII signaling has been shown to be important in driving tumor progression. Centrosomal protein 55 (CEP55), a member of the centrosomal relative proteins family, participates cytokinesis in the cell cycle. It exists in a few normal tissues and various tumor cells. The expression and function of CEP55 in human glioma cells need to investigate. In this study, the expression of CEP55 was detected in 40 cases of glioma tissues and 10 cases of non-tumor brain tissue. The proliferation of glioblastoma U251 cells was analyzed after transfection with EGFRvIII and CEP55 siRNA. We found that the expression of CEP55 was increased significantly in the glioma tissues than in normal brain tissue. The proliferation of U251 cells increased remarkably after transfection with EGFRvIII. Knockdown of CEP55 inhibited proliferation of U251 cells and was able to eliminate the effect of promoting proliferation induced by EGFRvIII in U251 cells. CEP55 played a key role in the proliferation of glioma cells and mediated EGFRvIII-stimulated proliferation in glioma cells. CEP55 might be a novel molecular therapeutic target in patients with gliomas expressing EGFRvIII.

Objective The expression of centrosomal protein 55 (CEP55) in human glioma cells was detected and the effect of CEP55 on proliferation and apoptosis of human glioma cell line U251 cells was analyzed.Methods The expression of CEP55 was measured by quantitative real-time PCR and Western Blot in 40 human glioma tissues of variable grades and 10 normal brain tissues.The prohferation of glioma cell line U251 was measured by CCK-8 kits and edu kits and the apoptosis of glioma cell line U251 was measured by flow cytometry after CEP55 silencing.Results The expression of CEP55 was increased significantly in 40 glioma tissues compared to 10 normal brain tissues (P ＜0.01).However,there was no statistical difference in the expression of CEP55 between patients with high grade glioma and lower grade glioma (P ＞ 0.05).The proliferation of U251 cells was inhibited markedly after CEP55 knockdown by RNAi (P ＜ 0.01);the apoptosis of U251 cells was profoundly increased after CEP55 knockdown (P ＜ 0.01).Conclusion The expression of CEP55 is significantly increased in human glioma cells compared with normal brain tissue;the overexpression of CEP55 promotes the proliferation of glioma U251 cells and inhibits the apoptosis of U251 cells.

BACKGROUND:Gliomas are the most common primary tumors in central nervous system. The prognosis of the patients with glioma is poor regardless of the development of therapeutic strategies. Its aggressive behavior mainly depends on the potent ability of proliferation. The transcription factor EGR1 (early growth response 1) is a member of a zinc finger transcription factor family which plays an essential role in cell growth and proliferation.METHODS:EGR1 expression levels in 39 glioma tissues and 10 normal brain tissues were tested by RT-qPCR and Western-blotting. The effects of EGR1 on U251 cells, U251 stem-like cells (GSCs), and U87 cells proliferation were assessed using in vitro and in vivo cell proliferation assays. The specific binding between EGR1 and CCND1 promoter was confirmed by CHIP assay. EGF was used to improve EGR1 expression in this assay.RESULTS:EGR1 expression levels in human gliomas are decreased compared with normal brain tissues, however, the patients with low EGR1 expression level showed significantly enhanced patient survival in all glioma patients. EGR1 silencing inhibited proliferation and induced G1 phase arrest in glioma cells. EGR1 contributed to proliferation by directly raising CCND1. Meanwhile, EGR1 overexpression induced by EGF was able to promote the proliferation of glioma cells.CONCLUSIONS:Our results show that stable knockdown EGR1 would inhibit glioma proliferation. The results suggest EGR1 showing lower expression in cancer tissues compared with normal tissues maybe still play an important role in tumor proliferation.