CLINICAL SIGNIFICANCE OF TELOMERASE ACTIVITY AND PERIPHERAL VENOUS BLOOD CK－20 EXPRESSION IN BLADDER TRANSITIONAL CELL CARCINOMA

A prospective study was performed in 91 patients with transitional cell carcinoma of bladder (TCCB) to detect the expression of CK-20 mRNA in peripheral blood and telomerase activity in tumor tissue. The relationship of the two as well as their clinical significance in the oncogenesis and development of TCCB was analyzed accordingly. PATIENTS AND METHODS Clinical Data Ninety-one patients with primary bladder carcinoma, 64 males and 27 females, aged 45-76y(median age 56.5y), we…     

Expression of a mutant hTERT in human bladder carcinoma cell line T24 and its clinical significance

Objective: To construct a mutant pEGFP- hTERT expression vector, to observe its steady expression in transfected human bladder carcinoma cell line T24 and its role in molecular regulatory mechanisms of telomerase, and to provide a new target gene for bladder cancer. Methods:PCR amplification was performed by using primers based on the known gene sequence of hTERT. PCR production was cloned into plasmid pGEMT-T easy and the sequence of mutant hTERT gene was analyzed. A recombinant mutant hTERT vector (pEGFP-hTERT) was constructed at the EcoR I and Sa/I sites of the pEGFP-C1 vector. After transfecting the fusion gene into bladder carcinoma cell line T24 by calcium phosphate-DNA coprecipitation, the steady expression of GFP-hTERT fusion protein was tested by fluorescent light microscopy. The proliferation changes of bladder carcinoma cell line T24 were detected by light microscopy and senescence correlated [3-galactosidase staining. Results: Identification of pEGFP-hTERT by enzyme digestion showed that mutant hTERT fragment had been cloned into EcoR I and Sal I sites of the pEGFP-C1 vector. The steady expression of GFP-hTERT fusion protein was localized in the nucleus of transfected cells. Expression of senescence-associated ~-galactosidase in transfected cells gradually increased with extended cultured time and cell growth was suppressed. Conclusion: The mutant-type hTERT gene suppresses the proliferation of bladder carcinoma cell line T24 by competitive effect on telomerase activity. This suggests that hTERT gene might be a suitable gene target for bladder cancer therapy.     

Plasmacytoid Transitional Cell Carcinoma of Bladder： A Clinico-pathological Study and Review of Literatures

Objective： To study the pathologic features of plasmacytoid transitional cell carcinoma of the bladder, and to analyze the diagnostic features, criteria for differential diagnosis and the clinical significance of the tumor. Methods： Two cases of bladder plasmacytoid transitional cell carcinoma were studied. Routine paraffin sections with HE staining, Pap smear and immunohistochemistry by S-P method were observed under a light microscope. Pathological and clinical data were analyzed by comparison with early reported cases in literatures. Results： A characteristic feature of this tumor was of deep invasion in the lamina propria and/or muscularis propria, in addition to the component of carcinoma in situ in the mucosa, when tumors were diagnosed. The histological pattern and cytological features showed similarity to a plasmacytoid tumor. The tumor cells were strongly positive for AE1/AE3, CEA and CK18. The prognosis appeared to be worse than ordinary transitional cell carcinoma. Conclusion： The plasmacytoid transitional cell carcinoma of bladder is rare but has typical pathological, immunohistological and clinical features. Pathologists should be aware of this kind of primary tumor of bladder.     

大剂量维生素 C 在肿瘤患者应用的安全性观察

Objective Dynamic detection of urinary Livin mRNA expression in patients with bladder cancer before and after operation and its clinical value in early diagnosis of bladder cancer. Methods Urine of 30 patients with initially diagnosed BTCC was collected before operation and one week, one month, three months, six months and 18 months after operation. Urine of 30 healthy volunteers and 30 Non-urological cancer patients was collected. Expression of survivin mRNA in urine exfoliated cells was detected by real-time PCR. Results Livin mRNA was highly expressed in the urine of 30 patients before operation, and the relative copy number was (96.33±35.79), and the expression level increased with the clinical stage and grade of the tumor from low to high; 2 of 30 patients in the control group showed high expression, the relative copy number was 43.17 and 47.52, the other was low expression, and the expression level was low. The copy number was (16.25±7.81); 30 cases in normal group were low expression, and the relative copy number was (13.74±1.57). The expression of Livin mRNA in urine of the case group was significantly higher than that of the control group and the normal group (P<0.05); the expression of Livin mRNA in urine of the case group was significantly lower than that of the control group (P<0.05); the expression of Livin mRNA in urine of the case group was significantly lower than that of the normal group (P<0.05); there was no significant difference between the control group and the normal group (P>0.05). Follow-up to 18 months after surgery, 5 patients with recurrence before reoperation (98.27±26.55) and 6 months after the initial operation were significantly different (P<0.05). Conclusion Dynamic detection of urinary Livin mRNA expression with high specificity and sensitivity can be used as an important noninvasive marker for early diagnosis of bladder cancer.     

Clinicopathological Significance of E-cadherin and PCNA Expression in Hunman Non-small Cell Lung Cancer

OBJECTIVE This study was designed to assess E-cadherin (E-cad)and proliferating cell nuclear antigen(PCNA)expression as well as their clinicopathological significance in hunman non- small cell lung cancers(NSCLCs).Possible molecular mechanisms of differentiation and metastasis of NSCLCs are discussed. METHODS Immunohistochemical and immunofluorescence double staining were performed to examine the expression of E-cad and PCNA in 68 primary NSCLCs cases. RESULTS The E-cad expression in squamous cell carcinomas and adenocarcinomas showed no significant difference.E-cad expression had a positive correlation with the histological- differentiated grade.A significant difference of E-cad expression was found between metastatic and non-metastatic groups.PCNA expression in squamous cell carcinomas and adenocarcinomas showed no significant difference.The PCNA expression had a reverse correlation with the histological-differentiated grade.A significant difference of PCNA expression was found between metastatic and non-metastatic groups.The E-cad and PCNA expression presented a reverse correlation. CONCLUSION E-cad expression is not associated with the histological type of NSCLC,but is associated with differentiation and metastasis of the cancer.Down-regulation of E-cad expression affects the proliferation of cancer cells.Conjoint analysis of E-cad and PCNA expression is a good way to evaluate tumor biological behavior.     

胃癌组织中Ets-1的表达及其与血管生成临床病理特征和预后的关系

Objective To investigate the expression of Ets-I in gastric carcinoma,pars-cancerous tissue and metastatic lymph nodes,and to determine the relationship between Ets-1 expression and clinicopathological features,angiogenesis and survival of patients with gastric carcinoma.Methods Gastric carcinoma tissue microarray was used to determine Ets-I protein expression by SP immunohistochemical staining in 189 advanced gastric cancer,54 papacancerous tissues,41 metastatic lymph nodes and 32 control tissues.Results The positive rates for Ets-1 expression of the carcinoma,paracancerous and control tissues were 71.4 %,29.6% and 18.8%,respectively,with a significant difference among the three groups(P ＜0.01).In the cancer tissues,the positive rate of Ets-1 protein expression was significantly associated with depth of invasion and lymph node metastasis(P ＜0.01),but not associated with degree of differentiation,Lauren's histological type,sex,age,and size of tumor(P ＞0.05).The positive rates for Ets-1 expression of the 41 gastric cancer and 41 metastatic lymph nodes were significantly different(P ＜0.05).In metastatic lymph nodes,the positive rate for Ets-1 expression was higher.The MVD in Ets-1 positive tumors was higher than that in the Ets-1 negative tumors,with a significant difference(P ＜ 0.05).Kaplan-Meier survival analysis showed that the survival time of Ets-1-negative patients was longer than that of Ets-1-positive patients (P ＜0.05).Cox regression analysis showed that Ets-1 expression was not an independent prognostic factor of gastric carcinoma.Conclusion A higher expression of Ets-1 is involved in carcinogenesis,development,invasion,and metastasis of gastric cancer.Ets-1 plays an important role in angiogenesis in gastric cancer.Ets-1 is a useful marker for predicting the outcome for patients with gastric carcinoma,though it is not an independent prognostic indicator.     

Expression of survivin and E-cadherin in breast cancer

Survivin is a member of the inhibitor of apoptosis (IAP) family, and is involved in the regulation of cell division. Survivin expression is related with poor survival of patients with tumors including neuroblastoma, non-small-cell lung cancer, breast carcinoma, gastric carcinoma, rectal cancer, recurrent ????????????colorectal carcinoma and bladder cancer. It is thought to be an important prognostic marker in cancers[1,2]. E-cadherin functionally belongs to transmembrane glycoproteins family,…     

Significance of the Expression of Rho-GDP Dissociation Inhibitorβ,γin Lung Squamous Cell Carcinoma and Adenocarcinoma

Objective:To study the expression of Rho-GDP dissociation inhibitor β,γ(Rho-GDIβ,Rho-GDIγ)in lung squamous cell carcinoma and adenocarcinoma and its relationship with the expression of RhoC(Ras homologus oncogenes C)and clinicopathologic parameters.Methods:Western blot assay was employed for Rho-GDIβ,Rho-GDIγ and RhoC in lung squamous cell carcinoma and adenocarcinoma and non-neoplastic lung tissues of 37 cases with fresh specimens.Results:The study showed that Rho-GDIβ,Rho-GDIγ and RhoC were expressed in lung cancer and non-neoplastic lung tissues,the level in lung cancer tissue was much higher than that in non-neoplastic tissues(P<0.001).In lung cancer,the expression of Rho-GDIβ was much higher in patients with lymph node metastasis(P=0.021),and the expression of Rho-GDIγ was much higher in poorly differentiated tumor than in well-differentiated and moderately differentiated tumor,but both of them were not correlated with other clinicopathologic parameters.The expressions of Rho-GDIβ and Rho-GDIγ were not correlated with the expression of RhoC.Conclusion:In lung cancer,Rho-GDIβ and Rho-GDIγ may play a role in the tumorigenesis,Rho-GDIβ may promote metastasis,and Rho-GDIγ may have some relationship with differentiation.     

Clinicopathologic observation of 6 cases with neuroendocrine carcinoma of the urinary bladder

OBJECTIVE To discuss the pathologic characteristics and the prognosis of neuroendocrine carcinoma of the bladder （NECB） in order to assist in making a precise diagnosis and to effectively treat the disease. METHODS Clinicopathologic features of the 6 cases with NECB were analyzed retrospectively. The personal data of the patients and the pathologic and immunohistochemical characteristics of the tumor were investigated. The follow-up of these patients was conducted over a time period ranging from 3 months to 9 years. RESULTS Transurethral resection of the bladder tumor was performed in all 6 patients. Based on the results of the pathologic examination, 4 of the cases were diagnosed as small cell neuroendocrine carcinoma and the other 2 were diagnosed as atypical carcinoid tumor. On immunohistochemical staining all tumors expressed neuroendocrine markers including neuronspecific enolase （CD56）, synaptophysin （Syn） and chromogranin （CgA）. In patients receiving partial cystectomy followed by postoperative chemotherapy, relapse was found in 4 of the 6 cases on follow up. CONCLUSION NECB is a rare entity and a tumor with high malignant potential and characteristic pathologic features. Hematuria is the cardinal symptom of NECB. Metastasis may occur at an early stage and the prognosis of the disease is poor. Final diagnosis of NECB depends on histopathologic examination and immunohistochemical assays. Surgical excision combined with radiotherapy and chemotherapy is considered an effective treatment.     

NUSAP1在膀胱尿路上皮癌中的表达及对预后评估的价值

目的:检测核仁和纺锤体相关蛋白1(NUSAP1)在膀胱尿路上皮癌中的表达及对预后评估的价值。方法:采用免疫组化方法检测NUSAP1在84例膀胱尿路上皮癌组织及10例正常膀胱上皮组织中的表达情况，并分析其表达与临床病理特征的关系；采用Western blot检测NUSAP1蛋白在T24和SV-HUC-1细胞系中的表达；采用单因素和多因素COX回归分析评价NUSAP1在膀胱尿路上皮癌中的预后价值。结果:免疫组织化学结果显示NUSAP1在膀胱尿路上皮癌中的阳性表达率为72.62%，在正常膀胱上皮组织中的阳性表达率为10.00%，差异有统计学意义（P<0.05）。NUSAP1在膀胱尿路上皮癌中的高表达与患者性别、年龄、肿瘤数目均无关(P>0.05)，而与肿瘤复发、病理分级、临床分期、转移有关(P<0.05)。NUSAP1蛋白在T24细胞系中的表达明显高于SV-HUC-1细胞系，差异有统计学意义(P<0.05)。NUSAP1表达、临床分期和转移均为膀胱尿路上皮癌的独立预测因素。此外，Kaplan-Meier曲线表明，NUSAP1阳性表达的膀胱尿路上皮癌患者无进展生存时间（PFS）明显少于NUSAP1阴性表达者。结论:NUSAP1在膀胱尿路上皮癌中呈现高表达，且与肿瘤复发、病理分级、临床分期、转移有关，NUSAP1是膀胱尿路上皮癌独立危险因素，其高表达提示预后较差，具有一定的临床价值。     

CIP2A在膀胱尿路上皮癌组织中的表达及其临床意义

目的 研究蛋白磷酸酶2A癌性抑制因子(cancerous inhibitor of protein phosphatase 2A,CIP2A)在膀胱尿路上皮癌组织中的表达及其与临床病理特征的关系,探讨其成为膀胱尿路上皮癌预后指标的可行性.方法 应用RT-PCR和Western blot检测CIP2A mRNA和蛋白在25例膀胱尿路上皮癌和对应癌旁组织中的表达情况;应用组织芯片技术和免疫组织化学方法,检测CIP2A在117例膀胱尿路上皮癌和30例癌旁组织中的表达情况,分析CIP2A与膀胱尿路上皮癌患者临床病理特征及预后之间的关系.结果 CIP2A mRNA和蛋白在25例配对膀胱尿路上皮癌组织中的表达水平明显高于癌旁组织.免疫组织化学检测发现,膀胱尿路上皮癌组织中CIP2A蛋白的阳性表达率为76.9％(90/117),明显高于癌旁组织的6.7％ (2/30),差异有统计学意义(P＜0.001).CIP2A表达与肿瘤病理分级(P＜0.001)、临床分期(P＜0.001)、肿瘤大小(P=0.002)和淋巴结转移(P=0.046)有关,但与年龄、性别及肿瘤数目无关(P＞0.05).KaplanMeier单因素分析显示,CIP2A蛋白高表达是总体生存率和无复发生存率的影响因素(P＜0.001).Cox多因素风险比例模型显示,与总生存率相关的独立预后因素为临床分期、肿瘤病理分级和CIP2A表达,与无复发生存率相关的独立预后因素亦为临床分期、肿瘤病理分级和CIP2A表达.结论 CIP2A蛋白在膀胱尿路上皮癌组织中高表达,可能与膀胱尿路上皮癌的进展有关,其表达状态是膀胱尿路上皮癌患者独立预后因素.     

膀胱病变组织中 ILK 激酶的表达及临床意义简

目的：探讨整合素连接激酶（integrin—linked kinase,ILK）在膀胱癌发生、发展中的作用。方法：采用免疫组化SP法,检测30例膀胱浸润性尿路上皮癌、25例非浸润性尿路上皮癌、20例膀胱内翻性乳头状瘤和5例正常膀胱黏膜组织中ILK激酶蛋白的表达。结果：浸润性尿路上皮癌ILK激酶阳性表达及阳性细胞率显著高于非浸润性尿路上皮癌、膀胱内翻性乳头状瘤和正常膀胱黏膜（P〈0．05）。结论：ILK激酶的表达水平与肿瘤发生、发展关系密切,高表达的ILK激酶预示着膀胱癌高度恶性、高侵袭性和预后不良。     

The plasmacytoid carcinoma of the bladder--rare variant of aggressive urothelial carcinoma

The WHO 2004 classification defines new histological and molecular variants of urothelial carcinoma. However, there are limited data available on the clinicopathological characteristics or prognosis of these variants. We present histopathological, molecular and clinical data of 32 plasmacytoid carcinomas of the bladder (PUC) showing that PUC is a high-grade tumor with molecular features of aggressive urothelial carcinoma, usually diagnosed in advanced pathological stage (64% pT3, 23% pT4) showing metastases in 60% of the patients. Average survival of our cohort of PUC treated with radical cystectomy and adjuvant chemotherapy was lower than what is typically seen for comparable conventional urothelial carcinomas. Eighty-seven percent of the PUCs showed a negative or strongly reduced membranous staining of E-cadherin. β-Catenin staining was negative in 22.5%, and 16.7% of the remaining tumors showed nuclear accumulation. Aberrant CK20 expression (negative or >10% of cells stained) and negative CK7 staining was found in 100% and 22.6%, respectively. Ninety-seven percent revealed positive staining for PAN-CK. CD138 was positive in 78%, whereas MUM-1 expression was negative in all cases. Multitarget fluorescence in situ hybridization showed all PUCs to be highly aneuploid and polysomic. Deletions on chromosome 9p21 seem to play an important role in this variant. FGFR3 and PIK3CA mutation analyses yielded no mutations in any of the PUCs analyzed. TP53 mutation analysis showed mutations in 29%. In summary, PUC is an aggressive variant of bladder cancer with molecular features of advanced bladder cancer and evidence of WNT pathway activation in some of the cases.     

沉默信息调节因子1在膀胱尿路上皮细胞癌中的表达与意义

目的:检测沉默信息调节因子1(SIRT1)在膀胱尿路上皮癌(BUC)中的表达,并探讨其与临床病理指标的相关性。方法:选取经临床病理诊断的BUC标本72例,其中低级别尿路上皮癌42例,高级别尿路上皮癌30例;阴性对照组为正常膀胱组织19例(膀胱镜检术或膀胱切开取石术中随机取活检时取得)。应用免疫组化方法(二步法)检测SIRT1分别在低级别与高级别BUC中的表达与分布,分析其与肿瘤的分期和病理分级的关系。结果:SIRT1在正常膀胱组织和膀胱癌组织中均有表达,正常膀胱组织中阳性表达率为31.58%(6/19),而在BUC组中,阳性表达率为77.78%(56/72),明显高于正常膀胱组织(P<0.05);并且随着肿瘤分期和癌理分级的升高SIRT1蛋白的表达增加,其中低级别膀胱癌阳性表达率为69.05%(29/42),高级别膀胱癌阳性表达率为90.00%(27/30),两者间的差异具有统计学意义(P<0.05)。结论:SIRT1在BUC中高表达,可能与BUC的发生、病理分级和临床分期有关,并有可能作为判断BUC侵袭力、监测复发的肿瘤标志物。阻断SIRT1表达有望成为膀胱癌靶向治疗的新靶点。     

复发性膀胱尿路上皮癌中IMP3 和CD44蛋白表达*

目的:评价IMP 3 和CD44蛋白在复发性膀胱癌中表达及二者的相关性。方法:收集2002年1 月至2012年12月经尿道膀胱肿瘤电切术诊断为尿路上皮癌（UC）的病例，其中筛选出6 个月内短期复发组25例和3 年以上较长期首次复发组29例。应用半定量免疫组织化学法检测短期复发组和较长期首次复发组UC病例中IMP 3 和CD44蛋白表达情况。结果:6 个月内UC复发为25例，6 例表达IMP 3，且均为高级别UC。3 年以上较长期首次UC复发为29例，仅有1 例低级别UC表达IMP 3。在短期复发组中IMP 3 阳性率为24%（6/ 25），表达强度为弱阳性16%（4/ 25）和强阳性8%（2/ 25），明显高于较长期首次复发组中阳性率3.45%（1/ 29）以及表达强度中的弱阳性3.45%（1/ 29）和强阳性0（0/ 29）。 CD44蛋白表达在两组之间差异无统计学意义。IMP 3 表达与UC复发患者的肿瘤分期、分级呈正相关，而CD44表达与肿瘤的分级呈负相关。IMP 3 表达与CD44表达之间无明显相关性。结论:IMP 3在UC短期复发组中的表达明显高于较长期首次复发组。IMP 3 可作为新的指标，并联合肿瘤病理分期、分级等因素对膀胱UC患者经尿道肿瘤电切术后短期复发的高危性进行预测。      

Reduced expression of metastasis suppressor RhoGDI2 is associated with decreased survival for patients with bladder cancer.

PURPOSE: RhoGDI2 was recently shown to be a metastasis suppressor gene in models of bladder cancer. We sought to further understand its importance in human cancer by determining the level of its expression and the distribution of its encoded protein in normal human tissues and cell lines and to evaluate whether its protein expression is a determinant of human bladder cancer progression. EXPERIMENTAL DESIGN: RhoGDI2 mRNA and protein expression was evaluated in cell lines and human tissues using Affymetrix and tissue microarrays, respectively. Tissue microarrays represented most human normal adult tissues and material from 51 patients that had undergone radical cystectomy for bladder cancer. In these 51 patients, the chi(2) test was used to test for associations between RhoGDI2 and stage, grade of urothelial carcinoma, histological type, and disease-specific survival status. Cox proportional hazards regression analyses were used to estimate the effect of RhoGDI2 expression level on time to development of metastatic disease and disease-specific survival time, adjusting for grade, stage, and histological type. RESULTS: In normal tissues, there was strong RhoGDI2 protein expression in WBCs, endothelial cells, and transitional epithelium. RhoGDI2 mRNA expression was inversely related to the invasive and metastatic phenotype in human bladder cancer cell lines. In patients with bladder cancer, univariate analysis indicated that reduced tumor RhoGDI2 protein expression was associated with a lower actuarial 5-year disease-free and disease-specific survival (P = 0.01). In addition, patients with tumors that had low or absent RhoGDI2 had a shorter time to disease-specific death (P 相似文献   

Hyperplasia and carcinomas in Pten-deficient mice and reduced PTEN protein in human bladder cancer patients

PTEN is a tumor suppressor gene mutated in many human cancers. We used the Cre-loxP system to generate an urothelium-specific null mutation of Pten in mice [FabpCrePten(flox/flox) (FPten(flox/flox)) mice]. Histologic examination revealed that all FPten(flox/flox) mice exhibited urothelial hyperplasia in which component cells showed enlarged nuclei and increased cell size. With time, 10% of FPten(flox/flox) mice spontaneously developed pedicellate papillary transitional cell carcinomas (TCC). This type of tumor also arose in FPten(flox/flox) mice treated with the chemical carcinogen N-butyl-N-(4-hydroxybutyl) nitrosamine. FPten(flox/flox) urothelial cells were hyperproliferative and showed increased activation of the survival signaling molecules Akt and extracellular signal-regulated kinase. In humans, 53% of primary bladder cancer patients exhibited decreased or absent expression of PTEN protein in either the cytoplasm or nucleus of tumor cells. In early bladder cancers, PTEN expression was repressed in 42% of superficial papillary TCC but in only 8% of cases of carcinoma in situ (CIS). In advanced bladder cancers, PTEN protein was significantly reduced (particularly in the nucleus) in 94% of cases, and this decrease in PTEN correlated with disease stage and grade. Thus, PTEN deficiency may contribute to bladder cancer both by initiating superficial papillary TCC and by promoting the progression of CIS to advanced invasive and metastatic forms.     

PRR11在膀胱癌组织中的表达及其对膀胱癌T24细胞增殖和凋亡的影响

目的 探讨富含脯氨酸蛋白11(PRR11)在膀胱癌组织中的表达及其基因沉默对膀胱癌T24细胞增殖和凋亡的影响.方法 采用免疫组织化学法检测57例膀胱尿路上皮癌组织及其癌旁组织中PRR11蛋白的表达,并分析PRR11蛋白表达水平与患者临床病理特征的关系.qRT-PCR和Western blot检测人永生化膀胱上皮细胞株S...