Stathmin基因在人成骨肉瘤细胞中的表达及意义

stathmin作为细胞信号转导分子在细胞分化及恶性肿瘤发生、发展上发挥重要作用。本研究旨在探讨Stathmin基因在人成骨肉瘤细胞中的表达,并探讨阻断其表达对该肿瘤细胞的生物学行为的影响。方法：RT-PCR及原位杂交法检测2个人成骨肉瘤细胞系及45例人成骨肉瘤组织中Stathmin基因表达情况;以高表达Stathmin基因的人成骨肉瘤细胞系SOSP-9607为靶细胞、反义Stathmin(ASODN)为阻断剂,通过MTT实验观察ASODN对该细胞系的生长抑制作用,并用流式细胞仪分析其对细胞增殖周期的影响。     

Evaluating the expression of CARMA3 as a prognostic tumor marker in renal cell carcinoma

Increased expression of CARMA3 has been reported to be involved in tumorigenesis and tumor progression of several cancer types. The aim of our study is to investigate the prognostic role of CARMA3 expression in patients with renal cell carcinoma (RCC). Real-time quantitative PCR was performed to detect CARMA3 mRNA expression level in 31 paired samples of RCC and adjacent noncancerous renal tissues. Subsequently, extensive immunohistochemistry was performed to detect CARMA3 protein expression in 114 RCC cases. Clinicopathological data for these patients were evaluated. The prognostic significance was assessed using the Kaplan–Meier survival estimates and log-rank tests. CARMA3 mRNA expression was significantly higher in RCC tissues compared with adjacent noncancerous renal tissues (3.525?±?1.233 vs. 1.512?±?0.784, P?<?0.001). In addition, high CARMA3 expression in RCC tissues was significantly associated with tumor size (P?=?0.026), histological differentiation (P?=?0.039), tumor stage (P?=?0.006), and the presence of metastasis (P?<?0.001). Moreover, Kaplan–Meier analysis showed that patients with high CARMA3 expression also had a significantly poorer prognosis than those with low CARMA3 expression (log-rank test, P?<?0.001). Furthermore, multivariate analysis illustrated that CARMA3 overexpression might be an independent prognostic indicator for the survival of patients with RCC. In conclusion, this work shows that CARMA3 may serve as a novel and prognostic marker for RCC and play a role during the development and progression of the disease.     

Overexpression of the stathmin gene in a subset of human breast cancer.

Stathmin is a highly conserved cytosolic phosphoprotein that destabilizes microtubules. Stathmin, which has been proposed as a relay protein integrating diverse cell signalling pathways, acts in vitro as a tubulin-sequestering protein, and its activity is dramatically reduced by phosphorylation. Interestingly, stathmin expression and phosphorylation are regulated during the control of cell growth and differentiation, and there is much evidence suggesting that in vivo stathmin plays a role in the control of microtubule dynamics during mitosis. Stathmin may thus be considered as one of the key regulators of cell division. We examined 50 human primary breast tumours for stathmin mRNA and protein expression and screened for abnormalities in the chromosome region harbouring the stathmin gene. Overexpression of stathmin was found in 15 tumours (30%). At the present stage, no clear correlation emerged between stathmin expression and several prognosis markers. Interestingly, perfect matching was observed between stathmin mRNA overexpression, protein overexpression and strong staining for stathmin on paraffin-embedded tumour sections when specimens were available. Furthermore, a tentative link between loss of heterozygosity (LOH) in the 1p32-1pter region and stathmin overexpression was observed. Our results suggest that stathmin might play a role in breast carcinogenesis and that stathmin-overexpressing tumours may represent a new subtype of breast cancer.     

ROR-γ在三阴性乳腺癌中的表达及临床意义分析

  目的  探讨视黄酸受体相关孤儿受体-γ（retinoic acid receptor-related orphan receptor-γ,ROR-γ）在三阴性乳腺癌（triple-negative breast cancer,TNBC）中的表达及临床意义。  方法  采用Oncomine网站数据库分析乳腺浸润性癌与癌旁组织中ROR-γ mRNA的表达差异,通过Kaplan-Meier Plotter网站评估ROR-γ mRNA在乳腺癌不同分子亚型中的表达预后意义。收集2011年1月至2014年12月收治于天津医科大学肿瘤医院的269例TNBC患者的组织标本和临床病理资料,采用免疫组织化学染色法（immunohistochemistry,IHC）检测TNBC组织中的ROR-γ蛋白表达,并探讨其与患者临床病理特征、预后的关系。  结果  Oncomine和Kaplan-Meier Plotter网站分析结果显示,乳腺浸润性癌组织中的ROR-γ mRNA表达显著高于癌旁组织（P<0.000 1）。相较于其他分子亚型,ROR-γ在TNBC中的预后意义更为显著,且与不良预后相关。IHC检测结果显示,ROR-γ蛋白在TNBC组织中的表达高于癌旁组织,ROR-γ在TNBC中的高表达率为18.2%（49/269）。ROR-γ高表达的患者具有较高的发病年龄（P=0.001）、已绝经状态（P＜0.001）、高肿瘤Ki-67指数（P=0.001）和淋巴结转移（P=0.003）。ROR-γ高表达是TNBC患者无病生存期（P=0.031）和总生存期（P=0.029）的独立影响因素。  结论  ROR-γ在TNBC组织中高表达,并与肿瘤的增殖活跃、淋巴结转移和患者不良预后相关,或可作为潜在的预后标志物,为TNBC的诊疗提供新的参考。      

Expression of CHD1L in bladder cancer and its influence on prognosis and survival

Chromodomain helicase/ATPase DNA-binding protein 1-like (CHD1L) is overexpressed and highly associated with poor prognosis in many malignancies. However, the role of CHD1L in bladder cancer (BC) has not been thoroughly elucidated. The aim of this study is to investigate the relationship of CHD1L expression with clinicopathological parameters and prognosis in BC. Immunohistochemistry was carried out to investigate the protein expression of CHD1L in 153 BC tissues and 87 adjacent noncancerous tissues. Our data found that CHD1L protein expression was significantly higher in BC tissues than in adjacent noncancerous tissues (P?<?0.001). CHD1L overexpression was significantly correlated with histologic grade (P?=?0.005) and tumor stage (P?=?0.009). The Kaplan–Meier survival analysis revealed that survival time of patients with high CHD1L expression was significantly shorter than that with low CHD1L expression. Multivariate analysis further demonstrated that CHD1L was an independent prognostic factor for patients with BC. In conclusion, CHD1L is likely to be a valuable marker for carcinogenesis and progression of BC. It might be used as an important diagnostic and prognostic marker for BC patients.     

Expression of NEDD9 in pancreatic ductal adenocarcinoma and its clinical significance

The aim of this study was to investigate the expression and prognostic significance of NEDD9 in pancreatic ductal adenocarcinoma (PDA). Expressional levels of NEDD9 mRNA and protein in paired pancreatic cancer lesions and adjacent noncancerous tissues were examined by quantitative real-time PCR and western blotting. NEDD9 expression was analyzed by immunohistochemistry in 106 patients with PDA. The correlations between NEDD9 immunostaining levels and clinicopathologic factors, as well as the follow-up data of patients, were analyzed statistically. NEDD9 protein and mRNA levels were elevated in pancreatic carcinoma lesions compared with the paired adjacent noncancerous tissues. A high level of expression of NEDD9 was significantly correlated with clinical staging (P?<?0.001), lymph node metastasis (P?<?0.001), and histological differentiation (P?<?0.001). Patients with a higher NEDD9 expression had a significantly shorter survival time than those patients with lower NEDD9 expression. The multivariate analysis revealed that NEDD9 could serve as an independent factor of poor prognosis. Our finding indicates that NEDD9 could be used as prognostic molecular marker and therapeutic target for PDA.     

Clinicopathological significance of SLP-2 overexpression in human gallbladder cancer

Several studies have indicated that overexpression of stomatin-like protein 2 (SLP-2) has been identified in several types of cancer. However, its role and clinical relevance in gallbladder cancer (GBC) is unknown. The purpose of this study was to reveal the prognostic significance of SLP-2 in GBC. The SLP-2 expression was examined at mRNA and protein levels by real-time quantitative polymerase chain reaction (qRT-PCR), and immunohistochemistry in GBC tissues and adjacent noncancerous tissues. Statistical analyses were applied to test the associations between SLP-2 expression, clinicopathologic factors, and prognosis. Immunohistochemistry and qRT-PCR showed that the protein and mRNA expression levels of SLP-2 were both significantly higher in GBC tissues than in adjacent noncancerous tissues. In addition, immunohistochemistry analysis showed that SLP-2 expression was significantly correlated with histological grade (P <0.001), pathologic T stage (P?=?0.019), clinical stage (P?=?0.001), and lymph node metastasis (P?=?0.026). The Kaplan–Meier survival curves indicated that patients with high expression of SLP-2 had shorter overall survival than those with low expression (P <0.001). Meanwhile, the Cox multivariate analysis indicated that high expressions of SLP-2 were an independent prognostic factor for patients with GBC. These data showed that SLP-2 may play an important role in human GBC tumorigenesis, and SLP-2 might serve as a novel prognostic marker in human GBC.     

Bmi-1 PTEN及E-Cadherin在结直肠癌中的表达和意义

  目的  检测结直肠癌组织中Bmi-1、PTEN和E-Cadherin的表达,探讨三者表达的相关性及意义。  方法  用Western blot检测Bmi-1在5对结直肠癌和癌旁活检组织中的表达,实时定量PCR法检测Bmi-1、PTEN和E-Cadherin mRNA在28对结直肠癌和癌旁活检组织的表达;用免疫组织化学法检测Bmi-1、PTEN和E-Cadherin在28例结直肠癌石蜡组织中的表达及其相关性。  结果  Bmi-1蛋白在5例结直肠癌活检组织中的表达明显高于癌旁组织;Bmi-1 mRNA在89.3%（25/28）的癌组织中的表达高于癌旁组织;PTEN和E-Cadherin mRNA在82.1%（23/28）的癌组织中的表达低于癌旁组织;统计学分析发现Bmi-1和PTEN及E-Cad? herin的mRNA表达水平呈负相关;28例结直肠石蜡组织中,Bmi-1、PTEN和E-Cadherin蛋白的阳性率分别为92.9%（26/28）、46.4%（13/28）和82.1%（23/28）,统计学分析发现Bmi-1和PTEN及E-Cadherin蛋白表达水平呈负相关。  结论  本研究发现Bmi-1和PTEN及E-Cadherin之间呈负相关,从组织水平证实Bmi-1对PTEN及E-Cadherin的调控关系,为Bmi-1作为预测结直肠癌发生及转移的分子标志物及治疗新靶点提供了新的理论依据。      

TC21/R-Ras 2在肝癌中的表达、亚细胞定位及意义

目的:探讨TC21基因在肝癌细胞、肝癌及癌旁组织中的表达、亚细胞定位及意义。方法:用RT-PCR检测TC 21在肝癌细胞系及人肝癌及癌旁组织中mRNA水平的表达,用免疫组化及Western印迹检测TC21蛋白在肝癌相关组织中的表达差异,用组织芯片技术结合免疫组化检测TC21蛋白的亚细胞定位。结果:RT-PCR检测结果表明TC21 mRNA在SMMC-7721、BEL-7402、Huh-7、Hep3B、HepG2、MHCC-97L、MHCC-LM3 7个肝癌细胞系及L-02和Chang liver 2株永生化人肝细胞系、7对肝癌及对应癌旁肝组织中均有较高表达;Western印迹结果表明上述细胞系中TC21蛋白表达与mRNA表达一致,TC21在4对肝癌及癌旁组织中均呈较高水平的表达;免疫组化检测结果表明TC21在人原发性肝癌、癌旁组织、硬化肝组织与正常肝组织中的表达阳性率分别为84.2%、77.2%、41.7%、0%,肝癌与肝硬化、正常肝相比显著高表达(P<0.05,P<0.01),与癌旁组织比无统计学意义(P>0.05);统计学分析结果表明TC21蛋白表达与肿瘤大小呈正相关(P<0.05),与是否肝硬化呈负相关(P<0.05);肝癌组织中TC21主要定位于肝癌细胞核,部分定位于胞质,膜定位不明显,TC21蛋白在肝癌细胞中的核定位显著高于癌旁肝细胞(P<0.001)。结论:本研究首次揭示TC21蛋白在肝癌组织中的高表达及核定位预示其在肝细胞恶性转化及肝癌发生发展中发挥重要作用。     

肝癌及相关组织中NET-1基因与蛋白的表达

背景与目的:NET-1是最近发现的肿瘤相关基因,它在肝癌中的表达目前尚未见报道。本研究在胚胎肝、成人肝、肝癌及相应的癌旁组织中筛检和比较NET-1基因和蛋白表达,以探讨NET-1基因在肝癌表达的特征。方法:用逆转录聚合酶链反应(RT-PCR)分别检测3例胚胎肝、4例成人肝、4例验证肝癌组织和28例肝癌与相应癌旁组织中NET-1mRNA的表达,由四星图像分析系统软件检测NET-1基因mRNA的光密度。用基因生物工程方法制备NET-1多克隆抗体,并用激光共聚焦显微镜观察免疫荧光细胞化学检测培养的人肝癌细胞系SMMC-7721中NET-1抗体的表达,免疫组化法检测28例肝癌与癌旁组织中NET-1蛋白的表达。结果:(1)正常成人和胎儿肝组织中NET-1基因mRNA不表达,4例验证肝癌组织中NET-1基因mRNA均呈阳性表达;肝癌与癌旁组织中NET-1基因mRNA高表达,阳性率均为85.71%(24/28)。肝癌组织中NET-1mRNA的平均光密度显著高于癌旁组织(0.64与0.47,P<0.05)。(2)经激光共聚焦显微镜观察SMMC-7721细胞中NET-1抗体的表达定位于胞浆。(3)免疫组化检测NET-1多克隆抗体在同组肝癌和癌旁组织中NET-1蛋白检出率分别为96.43%(27/28)和71.43%(20/28),两者差异具有显著性意义(P<0.05)。(4)RT-PCR方法和免疫组化方法检测肝癌和癌旁组织中NET-1基因mRNA表达与蛋白表达两者之间阳性率无显著性差异,并呈显著性正相关(癌中r=0.48,癌旁r=0.40,均P<0.05)。结论:NET-1基因表达可能是肝癌发生的早期分子事件,该基因在肝癌诊断中可能有一定的参考价值。     

Prognostic significance of USP22 as an oncogene in papillary thyroid carcinoma

Ubiquitin-specific protease 22 (USP22), a novel deubiquitinating enzyme, has been associated with metastasis, therapy resistance, and cell cycle progression. The purpose of this study was to investigate the expression level of USP22 in papillary thyroid carcinoma (PTC) samples and to evaluate its clinical significance in PTC patients. USP22 expression was examined in 30 fresh PTC tissues and paired adjacent noncancerous tissues by real-time quantitative RT-PCR. Immunohistochemistry for USP22 was performed on additional 156 PTC tissues. The clinical significance of USP22 expression was analyzed. We found that the expression levels of USP22 mRNA and protein in PTC tissues were both significantly higher than those in noncancerous tissues. Clinicopathological analysis showed that USP22 expression was significantly correlated with tumor size (p?=?0.036), extracapsular invasion (p?=?0.012), multifocality (p?=?0.014), lymph node metastasis (p?=?0.022), distant metastasis (p?=?0.005), and TNM stage (p?=?0.002). The Kaplan–Meier survival curves revealed that USP22 expression was associated with poor prognosis in PTC patients. USP22 expression was an independent prognostic marker of overall patient survival in a multivariate analysis. Our findings suggest that USP22 is an independent predictor of poor prognosis of PTC patients.     

Aberrant TIG1 methylation associated with its decreased expression and clinicopathological significance in hepatocellular carcinoma

Recently, it has been reported that tazarotene-induced gene 1 (TIG1) methylation was frequently detected in a variety of human cancers. However, the relationship between the TIG1 methylation and the characteristics of hepatocellular carcinoma (HCC) remains unknown. The aim of present study was to observe the promoter methylation of TIG1 in HCC tissues and assess its prognostic significance for HCC. Real-time quantitative polymerase chain reaction and methylation-specific polymerase chain reaction were used, respectively, to examine the mRNA expression and methylation status of TIG1 in 91 pairs of HCC and adjacent noncancerous tissues. The mRNA expression level of TIG1 was significantly lower in HCC tissues than in adjacent noncancerous tissues. The rate of TIG1 promoter methylation was significantly higher in HCC tissues than in adjacent noncancerous tissues (P?<?0.001). A strong correlation between downregulation and promoter methylation was found in these tumors (P?<?0.001). More importantly, TIG1 methylation status was related to tumor size (P?=?0.015), histological differentiation (P?=?0.004), and tumor stage (P?<?0.001). Kaplan–Meier survival analysis showed that TIG1 promoter hypermethylation was associated with a worse outcome in patients with HCC. Further, Cox multivariate analysis indicated that TIG1 methylation status was an independent prognostic factor for the overall survival rate of HCC patients. In conclusion, our data suggested that epigenetic silencing of TIG1 gene expression by promoter hypermethylation may play an important role in HCC.