Targeting Aurora kinase B attenuates chemoresistance in glioblastoma via a synergistic manner with temozolomide

BackgroundRecent studies have demonstrated that aberrant expression or activation of kinases results in oncogenesis of a wide range of cancers including GBM. Inhibition of kinases expression induces a reduction of therapy resistance. In this study, we investigate the underlying mechanism by which glioblastoma (GBM) cells acquire resistance to Temozolomide (TMZ) through Aurora kinase B (AURKB) thus to identify novel therapeutic targets and prognostic biomarkers for GBM.MethodsAURKB was identified as a key candidate kinase-encoding gene in chemoresistance regulation by using kinome-wide bioinformatic analysis. Afterwards, the potential biological functions of AURKB in oncogenesis and chemoresistance were investigated by lentivirus-dependent silencing of AURKB combined with qRT-PCR, western blot and in vivo intra-cranial xenograft mice models. Additionally, immunohistochemistry (IHC) assays were performed to explore the clinical significance of AURKB in glioma patients. Lastly, Chou-Talalay method was used to confirm the synergistic effect of TMZ combined with AURKB inhibitor.ResultsAURKB was among the most significantly up-regulated kinase-coding genes in TMZ resistant GBM cells according to database GSE68029, moreover, an increased expression of AURKB was closely associated with poor prognosis in glioma and GBM patients. AURKB knock-down resensitized U87 resistant cells to TMZ both in vitro and in vivo. Additionally, the combination of TMZ and Hesperadin, a specific AURKB inhibitor, significantly suppressed the proliferation of TMZ resistant GBM cells thus dramatically prolonged the survival of xenograft mice viaa synergistic effect with TMZ.ConclusionElevated AURKB expression was strongly correlated to TMZ resistant acquisition and poor prognosis, furthermore, targeting AURKB would be a potential therapeutic target for GBM patients.     

中性粒细胞、单核细胞和淋巴细胞对不同级别胶质瘤预后的评估价值

目的探讨术前外周血中性粒细胞与淋巴细胞比(neutrophil-lymphocyte radio,NLR)与单核细胞计数/淋巴细胞计数比(monocyte-tolymphocyte ratio,MLR)对不同级别胶质瘤患者预后的评估价值.方法本研究纳入2010年1月至2018年3月期间在哈尔滨医科大学附属第三医院神经外科接受手术的635名胶质瘤患者,其中低级别胶质瘤296例,高级别胶质瘤344例,收集其临床病历资料以及随访资料.通过R统计分析软件确定NLR、MLR对胶质瘤预后作用的最佳分界点,并以最佳分界点为界值对纳入研究对象进行分组.采用Kaplan-Meier生存分析、单因素和多因素Cox回归分析,评估术前外周血NLR、MLR对不同级别胶质瘤患者的预后的影响.结果高级别与低级别胶质瘤患者的术前NLR、MLR水平不同.NLR、MLR对胶质瘤预后影响的最佳分界点分别为1.83和0.16.在低级别与高级别胶质瘤中,NLR<1.83患者的中位生存期均大于NLR≥1.83患者,MLR<0.16患者的中位生存期均大于MLR≥0.16患者.在低级别胶质瘤中,NLR,MLR均是预后影响因素,其中高NLR是低级别胶质瘤预后的独立危险因素[OR=1.92(1.14~3.23),P<0.05];在高级别胶质瘤中,仅高NLR是胶质瘤患者预后的危险因素,且为独立危险因素[OR=1.42(1.05~1.93),P<0.05].结论NLR、MLR可以作为低级别胶质瘤患者预后的临床评估指标,仅NLR可作为高级别胶质瘤患者预后的临床评估指标.相比较MLR,NLR是更有效的胶质瘤患者预后的临床评估指标.     

Associations between clinical-pathological parameters and biomarkers,HER-2, TYMS,RRMI, and 21-gene recurrence score in breast cancer

BackgroundChemotherapy is the predominant treatment option for patients with breast cancer. Selection of patients according to biomarker will improve chemotherapy efficacy. In the present study, we examined the relations of the expression of candidate genes and 21-recurrence score (RS) results to patients’ demographic characteristics, histopathological factors, and outcomes.MethodsA total of 146 patients were enrolled in this study. The patients underwent 21-gene RS testing. In addition, expressions of candidate genes, TYMS, RRM1, TUBB3, TOP2A, PTEN, were detected. Information was obtained on age, tumor size, TMN stage, tumor grade, and status of Ki-67, HER2, ER and PR. The treatment information on the type of endocrine therapy was also obtained.ResultsResults clearly showed that the 21-gene RS significantly correlated with the TNM stage of breast cancer (P = 0.047). The RS also correlated with the number of sentinel lymph node (P = 0.038). The pathological type of tumors was strongly associated with the expression of RRM1 (P < 0.015), and slightly correlated with TYMS (P = 0.095) and tumor size (P = 0.061). Further analysis showed that TYMS and RRM1 were two independent factors affecting the disease progression of patients. Besides, for HER-2 stain, staining of grade 2 or above significantly increased the risk of disease progression.ConclusionOur studies showed that TNM stage and sentinel lymph node were important clinical parameters correlated with 21-gene RS results. Also, RRM1, TYMS and HER2 expressions were independent factors associated with disease progression in breast cancer patients. Future study is warranted to investigate the usefulness of these genes in treatment efficacy.     

RSK4: a new prognostic factor in glioma

Glioma is the most common and fatal brain tumour and has a poor prognosis. Ribosomal S6 protein kinase 4 (RSK4) has been found to be involved in multiple tumour types; however, the role of RSK4 in gliomas and its clinical relevance remain unclear. In the present study, RSK4 expression was found to be significantly increased in glioma tissues compared with matched adjunct non-noncancerous tissues. Moreover, the expression of RSK4 was significantly higher in high-grade (III and IV) glioma tissues than in low-grade (I and II) glioma tissues. The data showed that the expression of RSK4 was significantly correlated with WHO grade, three-year survival rate and five-year survival rate. Kaplan-Meier analyses showed that patients with high RSK4 expression had poor overall survival. In addition, multivariate Cox regression analysis showed that RSK4 might be an independent prognostic factor in glioma patients. Collectively, these results suggest that RSK4 may be a new prognostic factor in glioma patients, and RSK4 is expected to be a potential biomarker and a potential target for glioma therapy.     

Clinicopathologic significance of cystatin C expression in gliomas

Cathepsin B, one of the lysosomal cysteine proteases, has been related to tumor invasiveness. Cystatin C is the strongest inhibitor of cathepsin B. Knowledge of its participation in the progression of gliomas is limited. We investigated the expression of cystatin C and its association with the clinicopathologic features of 57 gliomas. Cystatin C and cathepsin B expressions were evaluated by immunohistochemical methods and by semiquantitative real-time polymerase chain reaction analysis for the corresponding messenger RNA. Disease-free survival was analyzed by the Kaplan-Meier method. Tumors with low cystatin C protein expression and high cathepsin B protein expression were significantly more likely to be of high grade, and this pattern was significantly correlated with high Ki-67 LI and tumor recurrence. Depressed expression of cystatin C messenger RNA in glioblastomas compared with low-grade astrocytomas was demonstrated. Multivariate analysis demonstrated high tumor grade, high Ki-67 labeling index, high cathepsin B expression, and low cystatin C expression correlated significantly with shorter disease-free survival. These results suggest that gliomas in patients with an unfavorable clinical outcome are characterized by depressed expression of cystatin C. Evaluation of cystatin C expression in gliomas provides useful clinical information, especially as a prognostic indicator.     

B7-H4在人胶质瘤组织中的表达及其临床意义

 目的： 探讨B7-H4在人胶质瘤组织中的表达及其临床意义。方法： HE染色分析了150例胶质瘤标本病理级别。免疫组化方法检测B7-H4在胶质瘤中的表达, 并分析其与临床病理参数及生存时间的关系。结果： HE染色检测150例胶质瘤标本病理级别,结果为I级12 例,II级50 例,III级39 例,IV级49 例。150例胶质瘤标本中97例B7-H4高表达,高表达率为64.7％,居于病理级别III～IV级;43例B7-H4低表达,低表达率为28.7%,居于病理级别I～II级。统计学分析提示组织标本中B7-H4的表达与年龄(P<0.01)和病理级别(P<0.01)存在明显相关,与肿瘤的部位没有显著相关性（P>0.05）。B7-H4阳性患者生存期更短（P<0.01）,多因素Cox 回归分析显示B7-H4、年龄、性别及病理级别均是胶质瘤患者的独立预后因素。结论： B7-H4在多数胶质瘤组织中表达较高,可作为脑胶质瘤患者的独立预后分子标志物和新的治疗靶点。     

Clinicopathological significance of Polo-like kinase 1 (PLK1) expression in human malignant glioma

Polo-like kinase 1 (PLK1), a variety of serine/threonine-protein kinase, has been reported to play important roles in malignant transformation. The purpose of this study was to investigate the clinicopathological significance of PLK1 expression in malignant glioma. A semi-quantitative RT-PCR assay was performed to detect the expression of PLK1 mRNA in 68 cases of glioma tissues and corresponding non-cancerous brain tissues. Additionally, the correlation of PLK1 mRNA expression with clinicopathological factors or prognosis of glioma patients was statistically analyzed. Multivariate analysis of prognostic factors was performed using the Cox proportional hazard model. Small interfering RNA was used to knockdown PLK1 expression in a glioma cell line and analyze the effects of PLK1 inhibition on growth, cell cycle, apoptosis and chemo- or radiosensitivity of glioma cells. Results showed that the expression of PLK1 mRNA was significantly higher in glioma tissues than in corresponding normal brain tissues. The expression of PLK1 mRNA was closely correlated with WHO grade, KPS and tumor recurrence of glioma patients (P = 0.022, 0.030 and 0.041, respectively). Meanwhile, the disease-free and overall survival rates of patients with high PLK1 mRNA expression were obviously lower than those of patients with low PLK1 mRNA expression. Multivariate analysis showed that high PLK1 mRNA expression was a poor prognostic factor for glioma patients (P = 0.028). The expression of PLK1 mRNA and protein was significantly down-regulated in stably transfected U251-S cells. PLK1 down-regulation could inhibit growth, induce cell arrest in G2/M phase of cell cycle and apoptosis enhancement in glioma cells. Further, PLK1 down-regulation could enhance the sensitivity of glioma cells to cisplatin or irradiation. Thus, the status of PLK1 mRNA expression might be an independent prognostic factor for glioma patients and targeting PLK1 could be a novel strategy for chemo- or radiosensitization of human malignant gliomas.     

Prognostic significance of CDC25B expression in gliomas

BACKGROUND: CDC25B is a cell-cycle regulatory protein, which is considered to be related to tumorigenesis and progression of tumours. AIMS: To elucidate the role of CDC25B in glioma, the expression of CDC25B and the association of the CDC25B expression with the clinicopathological parameters were investigated. METHODS: Fifty seven gliomas, which included 21 low-grade astrocytomas, 17 anaplastic astrocytomas and 19 glioblastomas, were studied. Protein expressions of CDC25B were evaluated by immunohistochemical methods. Semiquantitative and real-time RT-PCR analyses for the expression of CDC25B mRNA were also carried out. Disease-free survival (DFS) data were analysed by using the Kaplan-Meier method. RESULTS: High expression of CDC25B was identified in 18 of the 19 glioblastomas, in 10 of the 17 anaplastic astrocytomas, but not in any of the 21 low-grade astrocytomas. The CDC25B mRNA expression increased with the rise in histological grade. Increased CDC25B expression was correlated significantly with a shorter period of DFS, as shown by multivariate analysis. CONCLUSIONS: Patients with an unfavourable clinical outcome are characterised by the increased expression of CDC25B in their glioma samples. Useful clinical information, especially on its relevance as a prognostic indicator, is provided by the evaluation of CDC25B expression in gliomas.     

UBE2C is a marker of unfavorable prognosis in bladder cancer after radical cystectomy

It has been suggested that ubiquitin-conjugating enzyme E2C (UBE2C, also known as UBCH10) represents a promising cancer biomarker. However, the clinicopathological or prognostic significance as well as the functions of UBE2C in bladder cancer are largely unknown. To investigate the significance of UBE2C expression in bladder cancer, immunohistochemical analysis was performed using a tissue microarray. UBE2C positivity was observed in 51 of 82 (62%) bladder urothelial carcinoma cases treated with radical cystectomy. In contrast, UBE2C was negative in all of the non-neoplastic urothelium examined. UBE2C positivity was significantly associated with higher tumor stage (p=0.0061) and presence of lymphovascular invasion (p=0.0045). In addition, UBE2C positivity was significantly associated with shorter cancer-specific survival after cystectomy (log rank p=0.0017; multivariate hazard ratio, 2.49; 95% confidence interval, 1.09-5.71). Small interfering RNA-mediated suppression of UBE2C in UM-UC-3 bladder cancer cells inhibited cell proliferation in vitro. Taken together, our results suggest that UBE2C is a novel prognostic biomarker as well as a potential therapeutic target in bladder cancer.     

High expression of Anxa2 and Stat3 promote progression of hepatocellular carcinoma and predict poor prognosis

AimTo elucidate whether the interaction between Anxa2 and Stat3 could promote the progression of hepatocellular carcinoma (HCC) and that high co-expression of Anxa2 and Stat3 could predict poor prognosis in HCC patients.MethodsWe investigated Anxa2 and Stat3 expression using Western blot analysis in 4 HCC and adjacent nontumor tissues and using immunohistochemistry in 100 patients’ paraffin sections. Then we assessed the expression of Stat3, Anxa2 and co-expression of Stat3 and Anxa2 with relevant clinical pathological parameters and their prognostic value in HCC patients. The recurrence and overall survival rates were estimated using the Kaplan-Meier method and compared with the log-rank test. The prognostic analysis was carried out with univariate and multivariate Cox regressions models.ResultsThe incidence of high Stat3 expression in HCC tissues (35%) was significantly higher than that in non-HCC tissues (8%) (P < 0.001). The same result was observed in Anxa2 (P < 0.001). Also, the overexpression of Stat3 or Anxa2 showed a significant relationship with the recurrence of the 100 HCC patients (P = 0.012; P = 0.003). Additionally, tumor size >3 cm in diameter, multiple tumor number, and the presence of microvascular tumor thrombus were also significantly associated with recurrence in 100 patients. Then, all enrolled patients were divided into four groups according to IHC score of Stat3 and Anxa2, and the results indicated a significant difference in recurrence time between the subgroups (P < 0.001). What’s more, co-highexpression of Stat3 and Anxa2 was related to the presence of microvascular tumor thrombus (P = 0.003) and poor tumor differentiation (P < 0.001), but not relevant with other clinical features (All P > 0.05).ConclusionThe expression of Stat3, Anxa2, or co-high-expression of the two proteins was associated with HCC recurrence and survival.     

Bortezomib stabilizes mitotic cyclins and prevents cell cycle progression via inhibition of UBE2C in colorectal carcinoma

Substantial evidence implicates the ubiquitin-conjugating enzyme E2C (UBE2C) gene, in several human cancers, including colorectal carcinoma (CRC). We therefore investigated the prognostic value of UBE2C alterations in CRC and UBE2C signaling in CRC cell lines. UBE2C protein expression and UBE2C gene copy number were evaluated on clinical samples by immunohistochemistry and fluorescence in situ hybridization in a TMA format. The effect of the proteasome inhibitor bortezomib and small-interfering RNA knockdown was assessed by apoptotic assays and immunoblotting. UBE2C dysregulation was associated with proliferative marker Ki-67, accumulation of cyclin A and B1, and a poor overall survival. UBE2C expression was an independent prognostic marker in early-stage (I and II) CRC. UBE2C depletion resulted in suppression of cellular growth and accumulation of cyclin A and B1. In vitro, bortezomib treatment of CRC cells caused inhibition of cell viability via down-regulation of UBE2C. UBE2C knockdown by bortezomib or transfection with specific small-interfering RNA against UBE2C also caused cells to be arrested at the G2/M level, leading to accumulation of cyclin A and cyclin B1. In vivo, a significant reduction in tumor volume and weight was noted in mice treated with a combination of subtoxic doses of oxaliplatin and bortezomib compared with treatment with oxaliplatin or bortezomib alone. Altogether, our results suggest that UBE2C and the ubiquitin-proteasome pathway may be potential targets for therapeutic intervention in CRC.     

神经胶质瘤中β-抑制蛋白1的表达及意义

目的检测神经胶质瘤中β-抑制蛋白1(ARRB1)的表达水平及临床意义。方法采用RT-PCR和Western blotting检测胶质瘤细胞系中ARRB1的表达水平;通过肿瘤微阵列数据库(Oncomine)及Project Betastasis平台分析ARRB1 mRNA在神经胶质瘤中的表达情况;采用免疫组织化学方法检测神经胶质瘤组织及瘤旁脑组织中ARRB1的表达水平;应用Kaplan Meier分析ARRB1的表达水平与胶质瘤患者预后的关系。结果与正常人脑组织和人胚肾细胞系(HEK293)相比,神经胶质瘤细胞系中ARRB1的mRNA和蛋白水平降低;Oncomine数据库结果显示,多个神经胶质瘤基因芯片中ARRB1 mRNA水平较正常对照组明显降低(P0.001),ARRB1在恶性程度较高的胶质母细胞瘤中下降程度更加明显;免疫组织化学结果显示,与瘤旁脑组织相比神经胶质瘤中ARRB1表达降低,而且Ⅲ-Ⅳ期较Ⅰ-Ⅱ期神经胶质瘤降低更加明显。此外,Kaplan-Meier生存曲线结果显示,ARRB1的表达水平与神经胶质瘤患者的生存时间存在相关性(P0.05)。结论神经胶质瘤ARRB1水平下调,可作为反映神经胶质瘤临床病理学特点的指标;另外,ARRB1可作为判断神经胶质瘤患者预后的生物标志物。     

人脑胶质瘤组织中转化生长因子β1表达与Treg浸润的关系

目的:检测胶质瘤组织中转化生长因子β1(TGF-β1)表达,并分析其与CD4+ FOXP3+ 调节性T细胞(Treg)浸润的关系,探讨Treg浸润的临床意义.方法:采用免疫组织化学双标记的方法检测135例胶质瘤组织(WHO I 18,WHO Ⅱ45,WHO Ⅲ53,WH0Ⅳ19)、15例正常脑组织中TGF-β1、Tre...     

Overexpression of Arginase-1 is an indicator of poor prognosis in patients with colorectal cancer

AimArginase-1 (Arg-1) metabolizes l-arginine to l-ornithine and urea. It has been documented to have a role in various malignancies. However, the relationship between Arg-1 expression and clinicopathological characteristics of colorectal cancer (CRC) patients remains to be elucidated. The present study aimed to analyze the expression and prognostic value of Arg-1 in patients with CRC.Material and methodsThe mRNA and protein expressions of Arg-1 in fresh colorectal cancer tissue specimens and the corresponding noncancerous tissue specimens were examined by RT-qPCR (n = 24) and western blot analysis (n = 17). Arg-1 expression levels were determined in paraffin-embedded CRC tissue specimens (n = 236) by immunohistochemistry. The associations of Arg-1 expression and clinicopathological features and clinical prognosis in 236 CRC patients were analyzed.ResultsThe expression levels of Arg-1 were significantly higher in the CRC tissues compared with the matched noncancerous tissues, and elevated Arg-1 expression was remarkably associated with stage III-IV tumors (P = 0.007), lymph node metastasis (P = 0.019) and a plasma albumin concentration <35 g/l (P = 0.022). Kaplan-Meier analysis indicated that Arg-1 overexpression was associated with adverse prognoses for overall survival (OS) (P < 0.001) and disease-free survival (DFS) (P < 0.001) in all cases. Further analysis revealed that the patients with high Arg-1 expression had significantly shorter OS and DFS at the advanced stages (III + IV) (P = 0.032 for OS, and P = 0.012 for DFS) but not at the early stages (I + II) (P = 0.194 for OS, and P = 0.065 for DFS). Multivariate analysis revealed that Arg-1 overexpression was an independent prognostic factor for OS (P = 0.002) and DFS (P < 0.001) in patients with CRC.ConclusionThe data indicated that Arg-1 overexpression in CRC may be a marker that can discriminate subgroups of patients with a poor prognosis.     

CBX3在脑胶质瘤中的表达分析及其对胶质瘤细胞凋亡的抑制作用

目的 通过对胶质瘤公共数据库中CBX3表达量的分析,明确其与胶质瘤预后的相关性。流式细胞术以及对凋亡指标的检测分析其对胶质瘤细胞系凋亡水平的影响。 方法 通过对TCGA以及CGGA中胶质瘤患者基因芯片的数据进行分析,比较CBX3在不同级别胶质瘤中的表达量以及分析CBX3表达量与胶质瘤患者预后之间的关系。GO和Pathway分析对CBX3基因进行功能注释。最后在细胞系中探索CBX3基因对胶质瘤细胞系凋亡水平的影响。 结果 高级别胶质瘤中CBX3表达量明显高于低级别的胶质瘤中CBX3的表达量(P<0.01) 。高表达CBX3的患者预后比低表达CBX3的患者更差( P<0.05)。同时流式细胞术实验发现过表达CBX3可以显著降低胶质母细胞瘤细胞系U87MG和T98G的凋亡水平。 结论 CBX3的表达水平与胶质瘤的生存期呈负相关关系,高表达 CBX3可以降低胶质母细胞瘤细胞系的凋亡水平,故而CBX3或许可作为判断胶质母细胞瘤预后的有效指标。