Expression of HIF-1alpha and Glut-1 in human bladder cancer

HIF-1 is a heterodimer consisting of the HIF-1alpha and HIF-1beta subunits, and HIF-1alpha is the unique oxygen regulated subunit that determines HIF-1 activity. HIF-1alpha upgrades many gene products which include the glucose transporter protein 1 (Glut-1). Immunohistochemical studies using a monoclonal antibody specific for HIF-1alpha indicate that the overexpression of HIF-1alpha occurs in the most common forms of human cancer, including bladder cancer. The expression of Glut-1 in human bladder cancer is associated with poor prognosis and a low survival rate. To our knowledge, this is the first study to compare the expression of both HIF-1alpha and Glut-1 with clinicopathological characteristics in superficial and invasive human bladder cancer (all invasive bladder cancer patients received radical radiotherapy). The Kaplan-Meier survival analysis curve shows a significant association of HIF-1alpha expression with recurrence and survival in superficial bladder cancer and shows a significant association of Glut-1 with survival in invasive bladder cancer [chi2 (4)=10.52; Pr >chi2 =0.0012].     

Clinical significance and biological roles of CRKL in human bladder carcinoma

CRKL encodes an adaptor protein that has been recently reported to be overexpressed in various cancers and associate with the malignant behavior of cancer cells. However, the expression pattern of CRKL protein and its clinical significance in human bladder cancer have not been well characterized to date. In the present study, CRKL expression was analyzed in 82 archived bladder cancer specimens using immunohistochemistry, and the correlations between CRKL expression and clinicopathological parameters were evaluated. We found that CRKL was overexpressed in 31 of 82 (37.8 %) bladder cancer specimens. A significant association was observed between CRKL overexpression and tumor status (p?=?0.019). To further explore the biological functions of CRKL in bladder cancer, we overexpressed CRKL in BIU-87 and 5637 cell lines. Using CCK8 assay and colony formation assay, we showed that CRKL upregulation increased cell proliferation. In addition, transwell assay showed that CRKL could also facilitate invasion. Further study demonstrated that CRKL upregulation increased cyclin D1 expression and ERK phosphorylation. In conclusion, CRKL is overexpressed in bladder cancer and regulates malignant cell growth and invasion, which makes CRKL a candidate therapeutic target for bladder cancer.     

Expression of NRG1 and its receptors in human bladder cancer

Background:

Therapies targeting ERBB2 have shown success in the clinic. However, response is not determined solely by expression of ERBB2. Levels of ERBB3, its preferred heterodimerisation partner and ERBB ligands may also have a role.

Methods:

We measured NRG1 expression by real-time quantitative RT–PCR and ERBB receptors by western blotting and immunohistochemistry in bladder tumours and cell lines.

Results:

NRG1α and NRG1β showed significant coordinate expression. NRG1β was upregulated in 78% of cell lines. In tumours, there was a greater range of expression with a trend towards increased NRG1α with higher stage and grade. Increased expression of ERBB proteins was detected in 15% (EGFR), 20% (ERBB2), 41% (ERBB3) and 0% (ERBB4) of cell lines. High EGFR expression was detected in 28% of tumours, associated with grade and stage (P=0.05; P=0.04). Moderate or high expression of ERBB2 was detected in 22% and was associated with stage (P=0.025). Cytoplasmic ERBB3 was associated with high tumour grade (P=0.01) and with ERBB2 positivity. In cell lines, NRG1β expression was significantly inversely related to ERBB3, but this was not confirmed in tumours.

Conclusion:

There is a wide spectrum of NRG1 and ERBB receptor expression in bladder cancer. In advanced tumours, EGFR, ERBB2 and ERBB3 upregulation is common and there is a relationship between expression of ERBB2 and ERBB3 but not the NRG1 ligand.     

Clinical significance and biological roles of CARMA3 in human bladder carcinoma

Caspase recruitment domain and membrane-associated guanylate kinase-like domain protein 3 (CARMA3) was reported as an oncoprotein overexpressed in several cancers. The expression pattern of CARMA3 and its clinical significance in human bladder cancer have not been well characterized. In the present study, CARMA3 expression was analyzed in 90 archived bladder cancer specimens using immunohistochemistry, and the correlation between CARMA3 expression and clinicopathological parameters was evaluated. We found that CARMA3 was overexpressed in 35 of 90 (38.8 %) bladder cancer specimens. Significant association was observed between CARMA3 overexpression with tumor status (p?=?0.081) and tumor grade (p?=?0.027). To further explore the biological functions of CARMA3 in bladder cancer, we depleted CARMA3 in T24 and 5637 cell lines using small interfering RNA (siRNA). Using cell counting kit-8 (CCK8) assay and colony formation assay, we were able to show that CARMA3 depletion inhibited cell proliferation and colony number. Further study demonstrated that CARMA3 depletion decreased an expression of nuclear factor kappa B (NF-κB) targets cyclin D1 and Bcl-2 expression, as well as IκB phosphorylation. Luciferase reporter assay showed that CARMA3 depletion could downregulate NF-κB reporter activity. In conclusion, CARMA3 is overexpressed in bladder cancer and regulates malignant cell growth and NF-κB signaling, which makes CARMA3 a candidate therapeutic target for bladder cancer.     

Expression and function role of DNA methyltransferase 1 in human bladder cancer

BACKGROUND:

The identification of potential tumor markers will help improve therapeutic planning and patient management. Therefore, the aim of this study was to highlight the role of DNA methyltransferase 1 (DNMT1) in bladder cancer.

METHODS:

A total of 50 samples of nonmalignant urothelium, 65 of muscle‐invasive bladder cancers, 15 of distant metastasis, and 50 of nonmuscle‐invasive bladder cancers were selected for immunohistochemical staining analysis. Furthermore, human bladder cancer cell lines HT1376 and HT1197 were selected for cell and animal experiments investigating changes in tumor behavior, treatment response, and related signaling in bladder cancer.

RESULTS:

The incidence of nuclear DNMT1 immunoreactivity in the bladder cancer specimens (45%) was significantly higher than in nonmalignant urothelium (15%, P = .0005), and the incidence in cancer was positively linked to clinical stage (24% in ≤T1 vs 55% in T2‐T4, P = .0007). The staining of DNMT1 was also significantly linked to lower complete response rates (P = .0014) and reduced survival rates (P = .000). By in vitro and in vivo experiments, DNMT1 silencing vector reduced tumor growth and attenuated treatment resistance in bladder cancer cells. Less epithelial‐mesenchymal transition, less invasion, and slower tumor growth were noted in cancer cells with inhibited DNMT1. Furthermore, the epidermal growth factor receptor‐mediated phosphatidylinositol 3′–kinase‐protein kinase B pathway might be the mechanism underlying the effects of DNMT1 on bladder cancer.

CONCLUSIONS:

DNMT1 could be a significant clinical predictor for stage and treatment response of bladder cancer. Moreover, targeting this enzyme could be a promising strategy for treating bladder cancer, as evidenced by inhibited tumor growth and enhanced radiosensitivity. Cancer 2011;. © 2011 American Cancer Society.     

乳腺癌组织SATB1基因表达与临床病理特征的关系

目的通过检测核基质结合区结合蛋白质1(special AT-rich sequence binding protein 1,SATB1)在不同阶段乳腺癌组织中的表达水平,探讨其在乳腺癌发生发展中的作用及意义。方法收集西安交通大学第二附属医院2010-06-01-2012-04-01乳腺组织存档蜡块标本259例,采用免疫组化SP法测定SATB1的表达,采用χ2检验分析其与乳腺癌临床病理特征的关系。结果 SATB1蛋白在正常乳腺组织、乳腺囊性增生组织、乳腺不典型增生组织(癌前病变)、非浸润性癌(导管内癌、小叶原位癌)、早期浸润性癌(浸润性导管癌、浸润性小叶癌)及浸润性乳腺癌组织中的阳性率分别为6.2%(2/32)、6.4%(3/47)、20.4%(10/49)、45.0%(9/20)、52.9%(9/17)和76.6%(72/94),后三者的阳性率明显高于前三者,P<0.05;浸润性非特殊类型乳腺癌SATB1蛋白阳性率为88.5%(54/61),明显高于浸润性特殊类型乳腺癌的54.5%(18/33)和早期浸润性乳腺癌的52.9%(9/17),P<0.05;SATB1蛋白表达在分期晚或有淋巴结转移的乳腺癌中有升高的趋势,差异有统计学意义,P<0.05。结论 SATB1蛋白在乳腺癌发生和发展不同阶段的表达呈进行性上升的趋势。预示SATB1蛋白参与乳腺癌的发生和发展,有望成为乳腺癌早期预测及治疗的重要靶标。     

SATB2在乳腺癌组织中的表达及其对乳腺癌细胞生物学特性影响

【摘要】目的 观察乳腺癌组织中SATB2的表达情况及SATB2对乳腺癌细胞生物学特性的影响。方法 选择北京市隆福医院2017年1月至2018年6月收治的80例乳腺癌患者为研究对象,研究其新切除的乳腺癌组织及匹配的癌旁组织,分别从蛋白、RNA水平检测SATB2表达情况。在MCF 7细胞中转染SATB2,观察对MCF 7细胞增殖、迁移、侵袭的影响。结果 乳腺癌组织中SATB2阳性表达率高于癌旁组织(8125％ vs. 2500％,P<005)。Real time PCR显示,乳腺癌组织中SATB2 mRNA水平高于癌旁组织(15∶10,P<005)。Western blot检测结果显示,乳腺癌组织中SATB2蛋白表达水平高于癌旁组织(18∶10,P<005)。在MCF 7细胞中,转染pGV141 SATB2组细胞增殖率高于空白对照组及空载组(22∶10∶11,P<005)。MCF 7细胞中,转染pGV141 SATB2后迁移细胞数、侵袭细胞数均多于转染pGV141及空白对照组(45∶5∶6,28∶4∶6,P<005)。结论 乳腺癌组织中SATB2呈高表达,SATB2的高表达可促进乳腺癌细胞MCF 7增殖、迁移、侵袭。     

C-myc蛋白和增殖细胞核抗原在人膀胱癌组织中表达的意义

应用增殖细胞核抗原（PCNA）和C－myc单克隆抗体，以免疫组织化学方法检测正常膀胱组织和膀胱癌组织中Cmyc蛋白和PCNA的表达水平。结果表明：5例正常膀胱组织中未发现C－myc和PCNA阳性表达，阳性表达的C－myc和PCNA均定位于肿瘤的细胞核内。58例膀胱癌中C－myc和PCNA蛋白阳性表达分别为43．1％和62．1％，并且C－myc和PCNA阳性表达率与膀胱癌的病理分级、临床分期和患者术后生存率相关。提示C－myc和PCNA阳性表达与膀胱癌发生发展相关，并有可能成为评价膀胱癌预后的指标之一。     

SATB2在胃癌组织中的表达及临床意义

目的:探析SATB2在胃癌组织中的表达情况及临床意义.方法:选择2015年6月-2016年6月,在我院存档的胃癌手术切除样本共116例,采用免疫组化SP法对切片进行处理,根据着色程度及阳性细胞占比判断SATB2的表达状况,对患者进行随访,记录其生存期.使用χ2检验及Cox风险回归分析判断SATB2表达对胃癌患者预后的影...     

人膀胱癌中TGF-β1及其信号转导分子的表达及意义

目的探讨转化生长因子-β1(transforming growthfactorβ1,TGF-β1)及其超家族肽类的细胞内转导分子Smad4和Smad7在人膀胱癌中的表达及意义。方法采用免疫组织化学方法,对经过石蜡包埋的膀胱癌和正常膀胱组织进行定性和定位检测。结果Smad4在正常膀胱组织中的表达率较高,膀胱癌中较低,差异有统计学意义(P<0.05);TGF-β1、Smad7在正常膀胱组织中表达不明显或无表达,在膀胱癌中表达率较高,两者比较,差异有统计学意义(P<0.05);TGF-β1与Smad7的表达水平呈正相关(P<0.01);TGF-β1与Smad4的表达水平呈负相关(P<0.01)。结论TGF-β1和Smad7蛋白的阳性表达与膀胱癌发生、发展密切相关,参与膀胱癌恶性转化,而Smad4对膀胱癌的进展有抑制作用。     

TSHZ3在膀胱癌组织中的表达及其对膀胱癌细胞生物学功能的影响

目的:采用生物信息学分析TSHZ3基因在膀胱癌组织中的表达水平,并通过体外实验探讨TSHZ3基因对膀胱癌细胞增殖、迁移和侵袭的影响,以及对上皮-间质转化相关蛋白的调控作用。方法:运用Oncomine和GEPIA数据库分析TSHZ3基因在膀胱癌组织中的表达水平,利用HPA数据库分析TSHZ3蛋白在正常膀胱和膀胱癌组织中的表达情况,并收集膀胱癌标本验证TSHZ3蛋白的表达。在膀胱癌T24细胞中转染TSHZ3质粒,CCK8法和克隆形成实验检测过表达TSHZ3基因对T24细胞增殖的影响,Transwell实验检测过表达TSHZ3基因对T24细胞迁移和侵袭的影响,Western blot检测过表达TSHZ3对上皮-间质转化相关蛋白的影响。结果:Oncomine和GEPIA数据库显示TSHZ3基因在膀胱癌组织中的表达水平降低。HPA数据库表明TSHZ3蛋白在正常膀胱尿路上皮组织中呈中等强度染色,而在膀胱尿路上皮癌组织中呈弱-中等强度染色。免疫组化结果显示TSHZ3在癌旁组织中呈现高表达,在膀胱癌组织中呈现低表达。过表达TSHZ3显著抑制T24细胞的增殖、迁移和侵袭能力。过表达TSHZ3增加E-cadherin蛋白的表达,降低N-cadherin和Vimentin蛋白的表达。结论:TSHZ3基因在膀胱癌中低表达,可作为抑癌基因抑制膀胱癌的发展。     

乳腺癌组织HR与HER-2和SATB1表达相关性研究

目的：检测乳腺癌组织中激素受体（hormonereceptor,HR）、人表皮生长因子受体2（humanepidermalgrowthfactOrreceptor2,HER-2）和特异性核基质结合蛋白（specialATrichsequencebindingprotein1,SATB1）的表达,探讨HR与HER-2、SATB1及临床病理参数的关系。方法：应用免疫组织化学方法检测乳腺癌患者雌激素受体（estrogenre-ceptor,ER）、孕激素受体（progesteronereceptor,PR）、HER-2及SATB1蛋白的表达,荧光原位杂交方法检测HER-2基因扩增状态,统计学方法分析HR（包括ER及PR）的表达与HER-2、SATB1及临床病理参数之间的相关性。结果：乳腺癌组织ER及PR的表达均与患者年龄正相关（r=0．286,P=0．010;r=0．249,P=0．026）,ER的表达与肿瘤分级负相关（r=-0．306,P=0．006）;ER及PR的表达与肿瘤大小、组织学类型、淋巴转移情况及TNM分期均无明显相关性,P值均〉0．05。HR的表达与SATB1、HER-2及SATB1／HER2双阳性表达均呈负相关关系（r=-0．248,P=0．027;r=-0．392,P〈0．001;r=-0．150,P〈0．001）。结论：HR阳性患者治疗及预后相对较好;乳腺癌组织HR的表达与HER-2和SATB1呈负相关关系,三者之间可能存在相互联系的信号通路。     

膀胱癌组织中乳腺癌扩增基因1的表达及其意义

目的 探讨膀胱癌组织中乳腺癌扩增基因1（AIB1）基因的表达及其临床意义.方法 收集30例因膀胱癌行全膀胱切除术或经尿道膀胱癌电切术的标本,术前病理检查均为尿路上皮癌,采用免疫组织化学方法检测膀胱癌组织和癌旁组织中AIB1的表达情况.结果 膀胱癌组织AIB1的阳性表达率高于癌旁组织[43.3％（13/30）比3.3％（1/30）,x2=4.316,p＜0.05],高分期膀胱癌组织AIB1的阳性表达率高于低分期膀胱癌组织[T1、T2、L期分别为25.0％（2/8）、42.9％（6/14）、62.5％（5/8）,x2=4.623,P＜0.05].AIB1的表达与患者性别、年龄、肿瘤数目和病理分级无明显相关性（均P＞0.05）.结论 AIB1与膀胱癌关系密切,有可能成为判断膀胱癌预后的重要指标.     

Expression and biological role of cytoglobin in human ovarian cancer

Loss of cytoglobin is found to be involved in the progression of several human cancers. However, its expression pattern and biological roles in human ovarian cancers are not clear. In this study, we examined cytoglobin expression in 118 archived ovarian cancer specimens using immunohistochemistry. A total of 72 specimens (61.0 %) showed cytoglobin downregulation. cytoglobin downregulation positively correlated with advanced FIGO stage and tumor grade. Cytoglobin plasmid transfection was performed in SKOV3 cell line and siRNA knockdown was carried out in SW626 cell line. MTT, colony formation assay and matrigel invasion assay were carried out to assess the role of cytoglobin on cell proliferation and invasion. Cytoglobin overexpression inhibited cell growth, invasion, cell cycle progression and cyclin D1 expression in SKOV3 cell line and its depletion promoted cell proliferation, invasion, cell cycle transition and cyclin D1 expression. In conclusion, cytoglobin is downregulated in ovarian cancers and associated with advanced stage. Our data provides evidence that cytoglobin regulates the ovarian cancer cell proliferation and invasion.     

结肠癌组织中SATB1、FN1和PDGFRB蛋白表达水平及临床意义

目的:探讨结肠癌组织中特异AT序列结合蛋白1(special AT-rich sequence binding protein,SATB1)、纤维连接蛋白1(fibronectin 1,FN1)和血小板衍生生长因子受体（platelet-derived growth factor receptor,PDGFRB）蛋白表达水平及临床意义。方法:选取2012年1月至2012年12月在我院肛肠科住院的42例结肠癌患者组织作为病理,并选取癌旁组织（距肿瘤边缘＞5 cm）作为对照,用免疫组织化学法检测SATB1、FN1和PDGFRB蛋白的表达水平,分析它们与结肠患者临床病理参数间的关系及结肠癌患者术后生存期的关系。结果:SATB1、FN1和PDGFRB在结肠癌组织中阳性表达率分别为57.1%（24/42）、61.9%（26/42）和66.7%（28/42）,在癌旁组织中阳性表达率分别为9.5%（4/42）、7.1%（3/42）和9.5%（4/42）,差异有统计学意义（P＜0.05）;SATB1、FN1和PDGFRB表达水平与年龄和性别不相关;与TNM分期、肿瘤分化程度和有无转移相关;随访发现,13例结肠癌患者死亡,生存29例,中位生存时间为34个月;进行二分类Logistic逐步回归分析,结果显示,SATB1、FN1、PDGFRB、肿瘤浸润深度和有转移是结肠癌患者生存结局的独立危险因素。结论:SATB1、FN1和PDGFRB在结肠癌组织中高表达,与结肠癌的发生和发展密切相关,可作为判断患者预后的指标。     

miRNA-93在膀胱癌中的表达及其对T24细胞生物学特性的影响

目的：探讨miRNA-93在膀胱癌中的表达及对人膀胱癌细胞株T24细胞生物学特性的影响及其作用机制。方法：选取郑州大学附属南阳市中心医院泌尿外科2010年5月至2016年5月收治的79例膀胱癌患者病理组织标本及配对癌旁正常组织,通过qRT-PCR检测miRNA-93的表达水平。沉默miRNA-93后,采用CCK-8法、Transwell实验和流式细胞术分别检测细胞增殖和迁移能力的变化及细胞的凋亡情况;Western blotting检测AKT/p-AKT、GSK3β/p-GSK3β蛋白表达水平的变化。结果：miRNA-93在膀胱癌组织及癌细胞中高表达,且表达水平与癌灶大小、淋巴结转移、病理分级及T分期有关（均P<005）。沉默miRNA-93后,T24细胞的增殖能力显著降低（P<0.05）,沉默miRNA-93促进T24细胞的凋亡并抑制其迁移;同时,沉默miRNA-93后p-AKT和p-GSK3β的蛋白表达水平均显著下降（P<0.05）。结论： miRNA-93能够促进人膀胱癌细胞株T24细胞的增殖和迁移,并抑制凋亡,其作用机制可能与下调p-AKT、p-GSK3β蛋白表达有关,提示miRNA-93可以作为诊断和靶向治疗膀胱癌的潜在作用位点。     

新疆地区维吾尔族乳腺癌患者SATB1表达与临床病理特征的关系

目的 探讨维吾尔族乳腺癌患者组织中特异AT序列结合蛋白1（SATB1）的表达及其与临床病理特征的关系。方法 选择2014年1月至2015年1月新疆医科大学附属肿瘤医院收治的维吾尔族乳腺癌患者158例,其中原位癌26例,浸润性癌132例。采用免疫组化SP法检测乳腺癌组织中SATB1表达,并分析其表达与乳腺癌临床病理特征的关系。结果 SATB1在癌旁组织中的阳性表达率为8.0％（4／50）,原位癌组织中为42.3％（11／26）,浸润性癌组织中为70.5％（93／132）,差异有统计学意义（P＜0.05）。SATB1表达与乳腺浸润癌患者的原发肿瘤大小、淋巴结转移、TNM分期及组织学分级均有关（P＜0.05）,而与年龄、绝经情况、病理分型、激素受体及HER-2表达均无关 （P＞0.05）。结论 SATB1表达可能与维吾尔族乳腺癌的发生发展有关。     

Expression and regulation of tumor suppressor gene maspin in human bladder cancer

Maspin is a member of serine protease inhibitor family with tumor suppressing activity for breast and prostate cancers, acting at the level of tumor invasion and metastasis. However, there have been no published data regarding the role of maspin in human bladder cancer. We evaluated maspin expression in 65 series of bladder cancer samples (22 transurethral resection (TUR) and 43 radical cystectomy) and studied the regulatory mechanism of maspin gene activation in bladder cancer cells. Maspin expression was immunohistochemically detected in four (18.2%) patients with TUR and 22 (51.2%) patients with radical cystectomy whereas no expression was observed in normal transitional cells located at tumor-free area in bladder. The maspin expression was significantly correlated with the development of muscle invasive bladder cancer (P=0.00008). Using a luciferase reporter system, maspin promoter activity was induced in the maspin-positive bladder cancer cell lines as well as maspin-negative RT4 cells. Furthermore, treatment with the DNA methyltransferase inhibitor, 5-aza-2' deoxycytidine, and histone deacetylase inhibitor, trichostatin A, led to re-expression of maspin in RT4 cells. Our results indicate that maspin may contribute to bladder cancer development and that DNA methylation and histone deacetylation may be important for regulating maspin gene activation in bladder cancer cells.     

核转录因子κB在膀胱癌中的表达及临床意义

背景与目的：实体癌（如乳腺癌,肝癌,胰腺癌）中的研究提示核转录因子κB的异常表达对血管生长,细胞周期相关基因进行调控,本研究探讨膀胱癌及非癌膀胱粘膜中核转录因子κB及其调控基因的表达差异及其临床意义。方法：30例膀胱癌和13例非癌膀胱粘膜共43例冰冻切片行免疫组织化学检测p50,p52,p65,c-Rel,RelB,IκBa的蛋白表达水平,免疫印迹检测5例配对样本的p65(NF-κB家族一个重要亚基）的蛋白表达,13例配对标本RP－TCR检测p50,p52,p65,c-Rel,RelB,IκBa,周期蛋白D1,白介素－8的mRNA水平。结果：在膀胱癌组织及非癌膀胱粘膜中均见到p50,p52,p65,c-Rel,RelB,IκBa ,周期蛋白D－,白介素－8的mRNA表达,但前者较后者明显为强（分别为P＜0．01,P＜0．05,P＜0．01,P＜0．05,P＜0．05;P＜0．05,P＜0．05）,膀胱癌中p50,P52.P65,c-Rel,RelB核着色也明显增加（分别为P＜0．01,P＜0．01,P＜0．01,P＜0．01,P＜0．01）。同时,免疫印迹证明在膀胱癌中p65,核转录因子κB一个重要亚基的表达增加,膀胱癌中淋巴结转移组与无淋巴结转移组相比,p52,p65,c-Rel等蛋白表达等级计分有显著差异（分别为P＜0．01,P＜0．01,P＜0．05）.结论：膀胱癌中NF－κB家族及其调控基因表达较正常膀胱粘膜明显更强,并可能与膀胱癌淋纠转移相关。