粘着斑激酶在膀胱移行细胞癌中表达的意义

目的：探讨粘着斑激酶（FAK）在膀胱移行细胞癌（TCC）中的表达与肿瘤分化、浸润的关系,以及与p53、bcl-2、ki-67表达的关系。方法：采用免疫组织化学方法,测定FAK在81例膀胱TCC中的表达情况。结果：FAK在膀胱癌组织中的表面明显高于癌旁黏膜,浸润性膀胱癌明显高于非浸润性,p53或ki-67阳性膀胱癌明显高于阴性组。结论：FAK可能是一种肿瘤转化相关酶,又是一种肿瘤演变相关酶;它的表达量可作为膀胱癌发生发展过程中有价值的指标。     

膀胱癌中mdm2、p53蛋白和粘着斑激酶表达的病理意义

目的探讨mdm2、p53蛋白和粘着斑激酶(FAK)在膀胱移行细胞癌(TCC)中的表达与肿瘤生物行为的关系.方法采用免疫组织化学方法,测定mdm2、p53蛋白和FAK在81例TCC中的表达情况.结果mdm2、p53蛋白和FAK在TCC组织中的表达明显高于癌旁粘膜,mdm2在p53阳性TCC组中的表达明显高于阴性组.结论mdm2和p53在TCC发展过程中可能具有协同作用,联合检测mdm2、p53蛋白和FAK可作为TCC有价值的诊断指标.     

基质金属蛋白酶与膀胱癌分化和浸润的关系

目的探讨基质金属蛋白酶2(MMP-2)在膀胱移行细胞癌(TCC)中的表达与肿瘤分化和浸润,以及与p53和ki-67表达的关系.方法采用免疫组化DAKOEnVisionSystem方法,测定MMP-2、p53和ki-67在83例膀胱TCC中的表达.结果MMP-2在TCC中的阳性率为63.9%,表达显著高于癌旁黏膜(P＜0.01);MMP-2表达强度与TCC癌细胞的分化高低和浸润深度呈明显正相关(P＜0.05),并与ki-67表达有显著意义的关连(P＜0.01),但与p53表达无明显相关(P＞0.05).结论MMP-2在TCC的发展和浸润过程中发挥重要作用,可能成为预测TCC预后的参考指标,特别是与其他标记物联合使用.     

凋亡抑制基因bcl-2、p53及ki-67在膀胱良恶性病变中的表达

目的探讨凋亡抑制基因bcl-2、p53及ki-67在膀胱移行细胞癌(TCC)和腺性膀胱炎(CC)中的表达,及其与TCC分化和浸润程度的关系.方法采用免疫组化Dako Envision System方法,测定bcl-2、p53、ki-67在72例TCC和47例CC中的表达情况.结果Bc1-2、p53和ki-67在TCC中表达的阳性率分别为31.9%、65.3%和51.4%,在CC中表达的阳性率分别为89.4%、14.9%和10.6%;低分化TCC易于高表达p53和ki-67,ki-67高表达还与TCC浸润性显著相关.结论TCC的发生发展与bcl-2、p53基因调节失控有关bcl-2表达可能是TCC肿瘤发生过程中的早期事件,p53和ki-67高表达是TCC分化程度低及浸润性强的标志.     

基质金属蛋白酶与膀胱癌分化和浸润的关系

目的 探讨基质金属蛋白酶2（MMP－2）在膀胱移行细胞癌（TCC）中的表达与肿瘤分化和浸润，以及与p53和ki-67表达的关系。方法 采用免疫组化DAKO En Vision System方法，测定MMP－2,p53和ki-67在83例膀胱TCC中的表达。结果 MMP－2在TCC中的性率为63．9％，表达显著高于癌旁粘膜（P＜0．01）；MMP－2表达强度与TCC癌细胞的分化高低和浸润深度呈明显正相关（P＜0．05），并与ki-67表达有显著意义的关联（P＜0．01），但与p53表达无明显相关（P＞0．05）。结论 MMP－2在TCCR的发展和浸润过程中发挥重要作用，可能成为预测TCC预后的参考指标。     

Livinα在膀胱移行细胞癌组织中表达的研究

目的：研究膀胱癌相关Livin基因在膀胱移行细胞癌组织中的表达以及Livin基因表达与膀胱移行癌生物学行为的关系；研究抑癌基因p53在膀胱癌组织中突变以及Livin表达与p53基因突变的关系。方法：100例膀胱移行细胞癌组织，设计Livinα引物，以半定量RT—PCR的方法，检测膀胱癌组织LivinαmRNA，以免疫组化法检测癌组织中p53蛋白。将膀胱肿瘤病理分级、临床分期、是否复发等生物学行为及p53蛋白染色强度分别量化，与组织LivinαmRNA RT-PCR产物量进行等级相关分析。并以10例正常膀胱黏膜作为对照。结果：100例膀胱移行细胞癌，67例检测到LivinαmRNA表达，10例正常膀胱黏膜组织未检测到LivinαmRNA表达，LivinαmRNA在膀胱移行细胞癌组织中表达率高于正常膀胱黏膜，x^2=14．4，P=0．000。LivinαmRNA在膀胱癌组织中表达与p53蛋白阳性（rs=0．465，P=0．002）及肿瘤病理分级（rs=0．463，P=0．002）有相关性。结论：膀胱移行细胞癌组织中可见LivinαmRNA高表达，LivinαmRNA表达与膀胱癌病理分级及癌组织中p53蛋白呈正相关。     

Ki67、p53及EGFR在胃癌组织中的表达水平及临床意义

目的 探讨Ki67、p53及EGFR在胃癌组织中的表达水平及其临床意义.方法 收取行病理学检查的胃癌患者170例作为研究对象,对其胃癌组织及癌旁组织的石蜡切片进行ki67、p53及EGFR免疫组化染色.结果 胃癌组织中Ki67、p53及EGFR阳性表达率分别为71.18%、55.88%及45.29%;癌旁组织中Ki67、p53及EGFR阳性表达率分别为31.76%、12.94%及10.59%.胃癌组织阳性表达率均明显高于癌旁组织,差异具有统计学意义(P<0.05).3种指标均与肿瘤浸润深度、分化程度、淋巴结转移以及TNM分期密切相关,即随着肿瘤进展而其阳性率增高(P<0.05).Ki67与p53、EGFR均呈正相关(P<0.05);p53与EGFR相关性不显著(P>0.05).结论 Ki67、p53及EGFR与胃癌的发展关系密切,均可作为判断胃癌恶性程度及进展情况的可靠指标.     

检测基质金属蛋白酶及其抑制物对判断膀胱癌侵袭和转移的意义

目的 :研究基质金属蛋白酶 (MMPs)中MMP 2、MMP 9及MMP 9抑制物TIMP 1在膀胱移行细胞癌中的表达情况及其在判断膀胱癌侵袭转移中的作用。方法 :应用免疫组化LSAB法检测 90例膀胱移行细胞癌 (transitionalcellcarcinomaofbladder,TCCB)组织中MMP 2、MMP 9和TIMP 1蛋白表达水平。结果 :膀胱癌组织中MMP 2、MMP 9和TIMP 1蛋白表达显著高于正常膀胱黏膜。浸润性膀胱癌组织中阳性表达显著高于表浅性膀胱癌组织。阳性表达细胞在肿瘤 -间质界面较多。与病理组织学分级、分期均呈正相关 ;与肿瘤的复发和转移呈正相关。结论 :MMP 2、MMP 9和TIMP 1参与膀胱癌侵袭转移 ,在膀胱移行细胞癌中的表达具有预后价值。     

四种基因在单发和多发性膀胱癌组织中的表达及意义

目的探讨p16、p21、p53和上皮细胞膜抗原(EMA)在单发性和多发性膀胱癌组织中的表达及意义。方法收集临床正常、单发和多发性膀胱癌标本,通过免疫组化法检测膀胱癌组织中相关基因的蛋白表达和定位分布。结果与正常膀胱黏膜上皮相比,p16和p53蛋白定位于细胞核,随着膀胱癌恶性程度的升高和侵袭程度的增加,p16蛋白表达明显下降,而p53表达明显升高(P<0.05)。p21蛋白定位于细胞膜,高分化膀胱癌的p21蛋白表达低于低分化膀胱癌(P<0.05);多发性膀胱癌表达阳性率明显高于单发性(P<0.05)。EMA主要表达于细胞浆,随着肿瘤分级和(或)分期的增加,阳性细胞数逐渐减少(P<0.05),在单发和多发性膀胱癌中的表达差异有统计学意义(P<0.05)。结论 p16、p53、p21和EMA在膀胱癌组织中不同水平的表达与膀胱癌的分级/分期密切相关,对膀胱癌相关基因的检测有助于膀胱癌的临床诊断、预后判断和治疗方法的选择。     

宫颈腺癌HPV16／18感染与ki67、p53蛋白表达的关系

目的研究人宫颈腺癌组织中ki67、p53蛋白表达,分析HPV16／18感染与ki67、p53蛋白表达的关系。方法采用组织微阵列技术结合原位杂交和免疫组化（二步法）检测24例慢性宫颈炎和86例宫颈腺癌HPV16／18-E6DNA和ki67、p53蛋白表达情况。结果HPV16／18-E6DNA与ki67、p53蛋白表达在宫颈腺癌组织中的阳性率分别为65．1％、51．2％、45．3％,均显著高于慢性宫颈炎组织8．3％、0．0％、0．0％（P〈0．01）。HPV16／18感染与宫颈腺癌的病理分级和组织学类型无关,但与ki67表达呈正相关（P〈0．05）,与p53表达呈负相关（P〈0．05）。ki67、p53蛋白表达与宫颈腺癌的病理分级有关,G2、G3组阳性表达率均明显高于G1组（P〈0．05）。ki67、p53蛋白表达与宫颈腺癌组织学类型无相关性。结论宫颈腺癌的发生发展与HPV16／18感染及ki67、p53蛋白表达异常相关。     

Immunohistochemical findings of nitric oxide synthase expression in urothelial transitional cell carcinoma including dysplasia

Several in vitro studies have shown that nitric oxide (NO) produced by NO synthase (NOS), such as inducible NOS (iNOS) play an important role in tumor biology. We immunohistochemically examined the expression of iNOS and p53 proteins in patients with transitional cell carcinoma (TCC) of the urinary tract, including adjacent dysplastic lesions to determine the significance of the tumor behavior. Of total 94 tumors, in the present study, 41 (43.6%) tumors exhibited homogeneous immunostaining (diffuse strong positivity in tumor cells, >60%) and 53 (56.4%) tumors heterogeneous staining (variable positivity in tumor cells, 20-60%). No TCCs exhibited negative iNOS immunostaining was found. Thirty (31.9%) of 94 TCCs were positive with anti-p53 antibody, including 23 of homogeneous and 7 of heterogeneous staining. Of 23 TCCs with homogeneous p53 immunostaining, 11 tumors exhibited homogeneous iNOS immunoreaction. In the present study, dysplastic lesions adjacent to carcinomas were detected in 64 cases including 36 TCCs with homogeneous iNOS expression. All dysplastic lesions adjacent to the 36 TCCs with homogeneous iNOS immunostaining exhibited homogeneous iNOS immunostaining. No significant association between iNOS immunoreactivity and any clinicopathological factors as well as p53 immuno-reactivity were found. These in vivo findings provide evidence for frequent iNOS protein expression in TCC. In addition, our observations indicate that overexpression of iNOS expression may be one of the early events in the carcinogenesis of TCC.     

Different combinations of genetic/epigenetic alterations inactivate the p53 and pRb pathways in invasive human bladder cancers

Inactivation of both the pRb (pRb-cyclin D1/cyclin-dependent kinase 4/6-p16) and p53 (p53-p21(WAF1)-p14(ARF)) pathways is thought to be essential for immortalization in vitro and malignant transformation in vivo. We identified different combinations of pRb and p53 pathway alterations in 12 invasive transitional cell carcinomas (TCCs) and addressed the functional significance of the different combinations observed. Results showed four combinations of alterations including -pRb/-p53 (ie., pRb inactivated in the pRb pathway and p53 inactivated in the p53 pathway; four TCCs), -p16/-p53 (four TCCs), -p16/-p21(WAF1) (one TCC), and -p16/ -p14(ARF) (two TCCs). These groups include two new combinations (ie., -p16/-p53 and -p16/-p21(WAF1)) not reported previously for TCCs. An alteration in the key components of the p53 pathway was not detected in one invasive TCC that had inactivated p16. Note that all four TCCs with inactivated pRb had mutant p53; thus, the combinations of -pRb/ -p21(WAF1) and -pRb/-p14(ARF) were not observed. Only two of eight TCCs with altered p16 had concomitant p14(ARF) loss, demonstrating that simultaneous inactivation of these two 9p21INK4a tumor suppressor genes is not obligatory. To determine the biological phenotypes of TCCs with different combinations of pRb and p53 pathway alterations, their downstream responses to gamma radiation were studied in vitro. As expected, none of eight TCCs with mutant p53 responded to gamma radiation by elevation of p53, p21(WAF1), or mdm2 or by cell cycle arrest. Only two of four TCCs with wild-type p53 and wild-type pRb (the combination of -p16/-p14(ARF)) showed normal downstream responses to gamma radiation and underwent cell cycle arrest. Two TCCs with wild-type pRb and wild-type p53 (the combination of -pl6/-p21(WAF1) and one TCC with -p16) failed to show cell cycle arrest in response to radiation. This was attributed to the absence of p21(WAF1) in one TCC. In summary, these data support a model of invasive bladder cancer pathogenesis in which both the pRb and p53 pathways are usually inactivated and the biology of the tumor is impacted by the mechanism of their inactivations.     

Apoptosis in transitional cell carcinoma of bladder and its relation to proliferation and expression of P53 and Bcl-2

Transitional cell carcinoma of bladder (TCC) is a relatively common cancer among men. Tumor progression is associated with expression or modulation of several gene products that control apoptosis and proliferation. Apoptosis is a negative growth regulatory mechanism in tumors. The aim of this study is to examine apoptosis and related regulatory molecular markers in a group of patients with TCC. Paraffinembedded tissues from 49 patients with TCC were examined for the expression of bcl-2, p53 and Ki-67 by immunohistochemistry. Apoptosis was detected by TUNEL method. Correlation between apoptotic index (AI), proliferation index (PI) and bcl-2 and p53 expression with each other and with pathological grade was determined. Apoptosis was observed in 28.1% of TCC cases. The mean AI of all cases was 13.7+/-24. No correlation was found between apoptosis and differentiation status of carcinoma. Bcl-2 expression was weakly detected in only one sample. P53 expression was detected in 26 of cases with mean staining index of 102+/-96. A significant correlation between p53 and Ki-67 staining indices was observed (r=0.521, p=0.001). Both p53 and Ki-67 expression showed a good association with the pathological grade (p=0.0001 and p=0.004, respectively). None of the markers showed significant correlation with AI and no correlation was found between the ratio of AI to PI and other parameters either. In conclusion, the frequency of apoptosis in TCC of bladder appears not to be associated with tumor grade, and with bcl-2, p53 and Ki-67 expression.     

Human mismatch repair gene (hMSH2) product expression in relation to recurrence of transitional cell carcinoma of the urinary bladder

BACKGROUND: Several convincing studies have shown that the hMSH2 gene plays major roles in mismatch repair by recognizing mismatched bases and preventing mutations during DNA replication. Loss of this function may result in the accumulation of DNA replication errors or even the mutator phenotype (which may be responsible for the multiple mutations required for multistep carcinogenesis), and it has been found to affect the prognosis of patients. Thus, the authors felt that it would be of interest to study the expression patterns of hMSH2 protein in malignant tumors and to assess the correlation of hMSH2 protein to various clinical and pathologic features in these patients. METHODS: The authors examined the expression patterns of hMSH2 protein in 115 patients with transitional cell carcinoma (TCC) of the urinary bladder by immunohistochemical technique using monoclonal antibody to hMSH2 protein. RESULTS: The group in which hMSH2 was preserved (>20% of nuclei were positive for staining by anti-hMSH2 antibody) included 86 of 115 (75%) of the TCCs, whereas the remaining 29 cases (25%) belonged to the group in which hMSH2 was preserved (< or = 20% of tumor nuclei were positive), including 2 negative cases. Univariate analysis indicated that reduced expression of hMSH2 protein was significantly more frequent in high grade tumors than in low grade tumors (P = 0.04). Furthermore, statistical multivariate analysis revealed that hMSH2 reduction was significantly related to the recurrence (P = 0.02). CONCLUSIONS: These results suggest that abnormal expression of hMSH2 protein might be involved in tumorigenic processes and in the progression of some TCCs, and that the loss of hMSH2 protein expression might be a useful predictor of recurrence of TCC.     

Urothelium-specific expression of an oncogene in transgenic mice induced the formation of carcinoma in situ and invasive transitional cell carcinoma.

Although many genetic alterations are known to be associated with human transitional cell carcinoma (TCC) of the urinary bladder, relatively little is known about the roles of these molecular defects, singular or in combination, in bladder tumorigenesis. We have developed a transgenic mouse model of bladder tumorigenesis using a 3.6-kb promoter of uroplakin II gene to drive the urotheliums-specific expression of oncogenes. In this study, we demonstrate that transgenic mice bearing a low copy number of SV40T transgene developed bladder carcinoma in situ (CIS), whereas those bearing high copies developed CIS as well as invasive and metastatic TCCs. These results indicate that the SV40T inactivation of p53 and retinoblastoma gene products, defects frequently found in human bladder CIS and invasive TCCs, can cause the aggressive form of TCC. Our results also provide experimental proof that CIS is a precursor of invasive TCCs, thus supporting the concept of two distinct pathways of bladder tumorigenesis (papillary versus CIS/invasive TCC). This transgenic system can be used for the systematic dissection of the roles of individual or combinations of specific molecular events in bladder tumorigenesis.     

Is chromosome 9 loss a marker of disease recurrence in transitional cell carcinoma of the urinary bladder?

Investigation of transitional cell carcinoma of the urinary bladder (TCC) patients classified by recurrence and/or progression has demonstrated that loss of chromosome 9, as detected by FISH analysis of the pericentromeric classical satellite marker at 9q12, occurs early. A total of 105 TCCs from 53 patients were analysed in situ by two independent observers for loss of chromosome 9 using quantitative fluorescence in situ hybridization (FISH). All 53 primary tumours were evaluated for chromosomes 9, 7 and 17. Normal ranges for chromosomal copy number were defined for normal skin epidermis and bladder epithelium. Values for chromosome 9 copy number outwith the range 1.51-2.10 (mean +/- 3 x s.d. of normal values) were significantly abnormal. Twenty-five TCCs were detected with consistent monosomic scores. Of 89 TCCs, in which multiple tumour areas were analysed, 85 tumours (96%) demonstrated the same chromosome 9 copy number in all areas (2-6) analysed; only three tumours demonstrated heterogeneity for this locus. A total of 36% (12 out of 33) of patients with subsequent disease recurrence demonstrated loss of chromosome 9 in their primary and all subsequent TCCs analysed. Only a single patient (n = 20) with non-recurrent TCC showed loss of chromosome 9 (P = 0.0085). Of 53 primary tumours, eight showed significant elevation of chromosome 17. Of these patients, six demonstrated elevation in chromosome 7 copy number. No abnormalities were observed in non-recurrent patients. This study describes rapid quantitation of chromosomal copy number by FISH using a pericentromeric probe for chromosome 9 in TCC of the urinary bladder. Routinely fixed and processed material was evaluated without disaggregation. Strict quality control of FISH demonstrated that this technique was reproducible in a clinical environment and could be used to detect genetic changes relevant to patient outcome. It is proposed that loss of chromosome 9 from primary TCC of the urinary bladder identified patients at high risk of recurrence and possible progression.     

上皮细胞钙粘蛋白(E-cadherin) 在膀胱移行细胞癌中的表达及其意义

 目的　观察上皮细胞钙粘蛋白(E-cad)表达与膀胱移行细 胞癌(TCC)分级、分期等生物学行为的关系。方法　采用恒冷切片免疫组 织化学SP法,对28例TCC和10例正常膀胱粘膜新鲜冰冻标本进行E-cad表达的检测。结果　对照组10例正常膀胱粘膜E-cad均正常表达,实验组28例TCC,仅50%(14/ 28)正常表达,二组差异显著(P<0.01)。TCC不同分级、分期间E-cad表达有显著性差 异(P<0.05),G3比G1+G2表达低,浸润性TCC比浅表性TCC表达低。初发与复发、 单发与 多发TCC之间比较,E-cad表达均未见显著性差异(P>0.05)。本组未见E-cad表达完全 缺失的TCC。结论　正常膀胱粘膜组织E-cad保留表达,TCCE-cad表达降 低,E-cad表达与TCC分级、分期呈负相关;E-cad表达与TCC的数目,与肿瘤的初发或复发 无关。