表浅膀胱癌Pgp、突变P53蛋白、bcl-2表达的预测复发价值

目的探讨Pgp、突变P53蛋白、bcl-2表达在预测表浅膀胱癌复发方面的价值.方法应用免疫组化SP法,检测46例随访3年以上、术前未经化疗的原发性表浅膀胱移行细胞癌中Pgp、突变P53蛋白和bcl-2表达情况.结果Pgp、突变P53蛋白、bcl-2的阳性表达率分别为45.7%、58.7%、37.0%,单因素Kaplan-Meier生存分析示Pgp、突变P53蛋白、bcl-2表达与术后复发相关(P值均<0.01);多因素COX回归分析示术后复发与Pgp、突变P53蛋白表达有关(P值均<0.05),而与bcl-2表达无关(P值>0.05).结论Pgp、突变P53蛋白表达是判定原发性表浅膀胱移行细胞癌术后复发的独立预后指标.     

膀胱癌组织中MMP-2与TIMP-2 mRNA的表达及临床意义

目的 探讨基质金属蛋白酶-2(MMP-2)及其组织抑制因子-2(TIMP-2)mRNA在膀胱癌组织中的表达,以及与膀胱癌侵袭、转移和复发的关系。方法 应用半定量PCR和实时定量PCR(RQ-PCR)方法,检测10例正常膀胱组织和45例膀胱移行细胞癌组织中MMP-2和TIMP-2 mRNA的表达。结果 半定量PCR显示MMP-2、TIMP-2在膀胱正常组织中均呈阴性表达,而在45例膀胱癌组织中,MMP-2阳性率为66.7%(30/45),TIMP-2为57.8%(26/45)。定量PCR检测显示MMP-2表达量随肿瘤病理分级和临床分期的升高而增加,在复发病例中也增加,其组间差异有统计学意义(P＜0.05);TIMP-2的表达量随病理分级和临床分期的升高而降低,而且组间差异有统计学意义(P＜0.05),而在初发复发病例的组间差异无统计学意义(P＞0.05)。结论 MMP-2和TIMP-2对于膀胱癌的侵袭、转移和复发发挥了重要作用;MMP-2可能对判断膀胱癌的预后有一定的指导意义。     

Bcl-2 expression related to altered p53 protein and its impact on the progression of human pancreatic carcinoma.

p53 and Bcl-2 are two important factors related to apoptosis and tumorigenesis. In this study, a series of 52 cases of pancreatic carcinoma (PC) were investigated using an immunohistochemical assay to determine whether altered expression of Bcl-2 and p53 has an impact on the progression of this malignancy. Cytoplasmic immunoreactivity for Bcl-2 and nuclear staining of p53 was found in 12 (23.1%) and 32 (63.5%) cases of PC respectively. Furthermore, an inverse correlation between the expression of p53 and Bcl-2 existed in this series (P < 0.01). In a subgroup, the proportion of tumours showing that p53-positive and Bcl-2-negative staining was increased with increasing histological grade and clinical stage (P < 0.05), and moreover, the survival period of those patients whose tumour had this staining was shorter than those with other staining patterns of combined p53 and Bcl-2 (P < 0.05). Therefore, it is concluded that simultaneously aberrant expression of Bcl-2 and p53 may confer PC with more malignant clinicopathological characteristics.     

膀胱移行细胞癌P—gp表达与凋亡相关蛋白p53、bcl—2的关系

目的探讨膀胱移行细胞癌组织中MDR1基因产物P-gp表达与凋亡相关蛋白p53、bcl-2的相互关系及其临床意义.方法采用免疫组化S-P法检测107例原发性膀胱移行细胞癌组织中P-gp、p53、bcl-2蛋白的表达情况.结果P-gp、p53、bcl-2蛋白的阳性表达率分别为63.6%、72.9%、54.2%,三者的过度表达均与膀胱癌的病理分级、临床分期和术后腔内化疗后复发有关;P-gp表达与p53、bcl-2蛋白的表达密切相关(P＜0.01或P＜0.05).结论P-gp、p53、bcl-2蛋白的过度表达不仅是判定原发性膀胱移行细胞癌恶性程度和预后的重要指标,而且凋亡相关蛋白p53、bcl-2可能参与膀胱移行细胞癌多药耐药的形成.     

野生型PTEN基因高表达对膀胱移行细胞癌EJ细胞的抑癌作用

目的 探讨外源性野生型人酪氨酸磷酸酶（PTEN）基因的高表达对膀胱移行细胞癌EJ细胞的抑癌作用。方法 利用携带人PTEN基因的野生型、磷酸酶域突变型质粒体外分别转染人膀胱移行细胞癌EJ细胞。Western blot检测目的基因PTEN的表达,观察细胞形态变化及超微结构变化;MTIO法检测细胞增殖率及转染细胞对吡柔比星（THP）和丝裂霉素（MMC）的敏感性;Western blot法检测bcl-2蛋白的表达。以空载质粒作为对照。结果 质粒转染后,EJ细胞的PTEN蛋白表达上升75．0％。转染野生型质粒后,EJ细胞异型性低,出现典型凋亡小体,细胞增殖率下降40．1％,bcl-2蛋白表达被下调,并提高了对THP和MMC的敏感性。而转染突变型质粒的EJ细胞则无此作用。结论 野生型PTEN基因在体外对膀胱移行细胞癌EJ细胞增殖有明显抑制作用,诱导细胞凋亡,磷酸酶域突变型PTEN基因无此作用。野生型PTEN的抑癌作用可能与其对bcl-2蛋白表达的下调有关。     

Apoptosis, proliferation and p53, cyclin D1, and retinoblastoma gene expression in relation to radiation response in transitional cell carcinoma of the bladder

Purpose: To determine whether the apoptotic index, the Ki67 index, and the expression of the p53, cyclin D1, and retinoblastoma genes correlate with local control, overall survival, and time to distant metastases in invasive bladder cancer treated with external beam radiation.

Methods and Materials: Paraffin-embedded pretreatment biopsies from 83 patients with invasive transitional cell carcinoma of the bladder were scored morphologically for apoptosis and immunohistochemically for Ki67, p53, cyclin D1, and retinoblastoma gene expression. Survival analysis methods were used to assess overall survival, local control, and freedom from distant metastases. A multiple proportional hazard (PH) regression analysis was performed to study the prognostic value of the abovementioned biologic parameters (all divided into two categories, except Ki67) in addition to classical prognostic factors such as T stage, histologic grade, multifocality of the tumor, and completeness of transurethral resection. All patients were treated with external beam radiation as sole treatment. Median follow-up for the 19 patients still living was 7.5 years.

Results: Apoptotic index varied from 0% to 3.4% with a mean of 0.8% and a median of 0.6%. Ki67 index varied from 0% to 60% with a mean of 14% and a median of 12%. P53 protein was detectable in 61% of the tumors. Overexpression of cyclin D1 was observed in 39% of the tumors and loss of retinoblastoma protein in 23% of the tumors. High Ki67 index was found to be significantly associated with p53 expression (p = 0.04) and cyclin D1 overexpression (p = 0.023). Cyclin D1 overexpression was found more often in Rb-positive tumors than in Rb-negative tumors (p = 0.006). Other associations between the markers are less clear. Biologic markers were not correlated with T stage or grade. In the PH analysis local control was found to be significantly better for tumors with wild-type p53 (p = 0.028). Also, tumors with an apoptotic index above the median value (0.6%) had a significantly better local control rate (p = 0.035). Ki67 index (p = 0.35), retinoblastoma gene expression (p = 0.30) and cyclin D1 overexpression (p = 0.61) were not found to have an additional predictive value regarding local tumor control. None of the tested biologic parameters were found to be associated with overall survival. Time to distant metastases was significantly shorter for tumors with high Ki67 index (p = 0.01) and tumors with an apoptotic index less than median (p = 0.009).

Conclusions: The results of our study provide evidence for a prognostic value of p53 expression and apoptotic index with respect to the radiation response in bladder cancer in addition to more conventional prognosticators. The value of these parameters as a predictive assay for radiation response warrants confirmation in larger and prospective studies.     

Caspase-3与Bcl-2蛋白在膀胱移行细胞癌中的表达及意义

背景和目的:肿瘤的发生是细胞异常增生和凋亡不足的结果。以往研究认为凋亡的发生与Caspase-3和Bcl-2蛋白有密切的联系。本研究拟通过检测Caspase-3和Bcl-2蛋白在膀胱移行细胞癌中的表达,探讨其在膀胱移行细胞癌发生、发展中的意义。方法:采用免疫组织化学(SP)法,对52例膀胱移行细胞癌组织和10例正常膀胱粘膜组织中的Caspase-3及Bcl-2蛋白表达情况进行检测。结果:膀胱移行细胞癌组织中的Caspase-3蛋白表达率(53.8%,28/52)明显低于正常膀胱粘膜组织(90.0%,9/10),Caspase-3的蛋白表达与膀胱移行细胞癌的病理分级有关(P<0.05),但与临床分期没有明显的联系;膀胱移行细胞癌组织中的Bcl-2蛋白表达率(51.9%,27/52)明显高于正常膀胱粘膜组织(20.0%,2/10),且Bcl-2蛋白表达情况与病理分级、临床分期均无关(P>0.05)。在正常膀胱粘膜中Caspase-3与Bcl-2的蛋白表达呈负相关(rs=-0.659,P<0.01)。结论:Caspase-3蛋白的高表达与Bcl-2蛋白的低表达在膀胱癌的发生和细胞凋亡的调控方面可能起着重要作用。     

NF-κBp65、Bcl-2、Bcl-xL蛋白在胰腺癌中的表达及其与P53和凋亡指数的关系

背景与目的:核因子κB通过调控下游bcl-2家族基因表达参入细胞凋亡调节过程,而p53除了参入细胞周期调节外也参入依赖bcl-2基因的细胞凋亡调控。在胰腺癌中NF-κBp65、Bcl-2和Bcl-xL蛋白表达与p53和凋亡的关系还不清楚。本研究系统地分析了核因子κBp65及其下游的bcl-2和bcl-xL抑凋亡基因在胰腺癌(PC)中的表达以及与P53蛋白表达和凋亡指数(AI)的关系。方法:免疫组化法检测25例胰腺导管腺癌(PC)和9例正常胰腺组织(NP)中P53蛋白表达;Western印迹法分析NF-κBp65蛋白表达;RT-PCR分析Bcl-2和Bcl-XL蛋白表达,TdT酶介导的原位缺口标记(TUNNEL)法了解胰腺癌细胞凋亡情况(AI)。结果:P53在PC组织阳性率(56%,14/25),高于NP组织阳性率(P<0.00);NF-κBp65、Bcl-2和Bcl-xL在PC组织表达相对值分别为:1.06±0.16、0.79±0.13、0.76±0.24,分别高于NP组织:0.23±0.016、0.23±0.074、0.18±0.026(分别P<0.05);14例p53阳性PC中,NF-κBp65、Bcl-2和Bcl-Xl表达相对值分别为:1.32±0.23、0.92±0.33、0.82±0.21;11例p53阴性PC中,NF-κBp65、Bcl-2和Bcl-xL表达相对值分别为:0.78±0.15、0.54±0.19、0.71±0.28(分别P<0.05,P<0.05,P>0.05);NF-κBp65和Bcl-2分别与P53表达有明显的正相关性(分别P<0.01,P<0.05),Bcl-xL与P53无相关性(P>0.05),NF-κBp65与Bcl-XL表达正相关(P<0.01),而NF-κBp65与Bcl-2表达无相关性(P>0.05);胰腺癌平均AI为(15.4±6.48)%,NF-κBp65表达与AI负相关(r=-0.297,P<0.05),而Bcl-2、Bcl-xL与AI无相关性(r=-0.203,P>0.05;r=-0.156,P>0.05)。结论:胰腺癌中抗凋亡因子表达上调,凋亡指数主要决定于NF-κBp65蛋白水平;NF-κBp65通过对下游bcl-xL基因表达的调控参与胰腺癌凋亡过程;p53在凋亡调节过程中起重要作用。     

AgNORs数量的变化在动脉化疗配合手术治疗膀胱癌中的意义

目的通过AgNORs染色分析技术检验膀胱癌动脉化疗前、后Ag,NORs均值的变化,评价膀胱癌术前行动脉化疗的意义.方法25例膀胱癌患者,经术前膀胱镜活检证实为膀胱移行细胞癌(TCC),其中Ⅱ级8例,Ⅱ～Ⅲ级4例,Ⅲ级8例,Ⅲ～Ⅳ级1例,Ⅳ级4例.经腹壁下动脉插导管至髂总动脉行ADM+CDDP联合化疗1～2个疗程后行膀胱部分切除术.将膀胱镜及手术所取之病理标本经Ag,NORs技术染色,计数其细胞核内平均AgNORs数量,进行配对t检验,并对患者进行术后随访.结果配对t值为5.230,而t(0.01)24=2.787,t＞t',所以P＜0.01,化疗前后膀胱癌组织AgNORs均值间有极显著差异.25例患者无明显并发症.经随访3年复发率为8.33%,5年复发率为12.50%,5年存活率为87.50%.结论术前经动脉化疗可降低膀胱癌的恶性程度,降低复发率,提高生存率,其效果优于单纯膀胱部分切除术+术中黏膜下注射抗癌药物,对恶性度高的膀胱癌是一种理想的治疗方法.     

MMP2和E-cadherin表达在膀胱癌中的侵袭效应

目的　探讨基质金属蛋白酶 2MatrixMetalloproteinase 2 (MMP2 )和钙粘附分子E cadherin(E CD)在膀胱移行细胞癌表达及侵袭效应。方法　应用S -P免疫组化染色法 ,检测 10例正常膀胱粘膜 ,96例膀胱移行细胞癌MMP2和E CD表达情况。结果　正常粘膜MMP2呈阴性表达 ,在膀胱移行细胞癌Ⅰ级、Ⅱ级、Ⅲ级阳性表达分别为2 3 % (7/3 0 )、5 8.3 % (2 1/3 6)、76.7% (2 3 /3 0 ) ,三组之间差异显著 (P <0 .0 1) ,正常粘膜E CD呈强阳性表达 ,膀胱移行细胞癌Ⅰ级、Ⅱ级、Ⅲ级正常表达分别为 86.7% (2 6/3 0 )、5 2 .8% (19/3 6)、2 6.7(8/3 0 ) ,三组之间差异显著 (P <0 .0 1)。结论　MMP2阳性表达和E CD的异常表达与膀胱移行细胞癌病理分级、浸润深度及生存率密切相关 ,检测MMP2和E CD的表达有助于判定膀胱移行细胞癌侵袭能力。     

环氧化酶-2 mRNA在膀胱癌中的表达及意义

目的 :探讨环氧化酶 -2 (COX 2 )mR NA在膀胱移行细胞癌 (TCCB)组织中的表达及其意义。方法 :采用RT PCR方法检测 78例TCCB组织中COX 2的表达 ,16例正常黏膜作为对照。结果 :16例正常组织无COX 2表达。COX 2表达阳性率为 3 5 9% ( 2 8/78) ,COX 2表达与TCCB分级、分期之间呈正相关 ,P <0 0 5。COX 2的表达在复发者中的表达率高于未复发者 ,P <0 0 5。COX 2的表达在转移者中的表达率高于未转移者 ,P <0 0 5。结论 :COX 2在TCCB的发生发展中起重要的正性调节因子的作用。COX 2与TCCB的恶性程度和进展恶化有关。COX 2可作为判断TCCB预后的指标。     

P33ING1、p53在膀胱移行细胞癌中的表达及与细胞凋亡的相关性

目的:探讨抑癌基因P33ING1在膀胱移行细胞癌(BTCC)中的表达及其与p53蛋白表达及细胞凋亡的相关性.方法:利用免疫组化S-P法和TUNEL.法检测83例BTCC及11例正常膀胱黏膜组织P33ING1、p53的表达及细胞凋亡指数(AI).结果:83例膀胱移行细胞癌组织中,P33ING1蛋白的阳性表达率为59.03%,而正常膀胱黏膜组织中P33ING1蛋白阳性表达率为90.9%.P33ING1蛋白表达与膀胱移行细胞癌的WHO肿瘤分级有相关性.spearman相关分析表明P33ING1蛋白表达与p53蛋白表达正相关(P<0.05).AI与P33ING1及p53蛋白表达无相关性.结论:P33ING1在膀胱移行细胞癌中表达下降可能在膀胱移行细胞癌的发生、发展过程中起重要作用,P33ING1与p53基因具有协同作用,同时检测p53的状态和P33ING1表达水平,对于膀胱癌的诊断、治疗和预后判断可能具有积极意义.     

膀胱癌细胞凋亡、P-gP、MDM2、p53基因表达的意义

目的:研究膀胱癌细胞凋亡、P-gp、MDM2、p53基因表达的意义。方法:采用免疫组化法及TUNEL法检测11例正常膀胱黏膜和83例膀胱移行细胞癌中P-gp、MDM2、p53基因的表达及细胞凋亡指数(AI)。结果:83例膀胱移行细胞癌中P-gp阳性71.08%,p53阳性50.6%,MDM2阳性59.03%,AI=1.4335±0.3863。MDM2阳性表达与病理分级及临床分期呈负相关(r=-0.263,P=0.016及r=-0.388,P=0.001);p53阳性表达与病理分级及临床分期呈正相关(r=0.492,P=0.001;r=0.341,P<0.01);P-gp阳性表达与病理分级及临床分期无相关性;P-gp、MDM2、p53 3种蛋白间表达无相关性;AI与MDM2呈负相关(r=-0.226,P=0.042);AI与病理分级及临床分期呈正相关(r=0.642,P=0.000;r=0.455,P=0.000);AI与P-gp、p53无相关性。AI、p53、MDM2的表达与5年生存率有关。结论:检测P-gp可指导膀胱癌化疗药物的选择;联合运用临床分期及病理分级并检测p53、MDM2可用于判断膀胱移行细胞癌的预后。     

Apoptosis and p53, Bcl-2 and bax gene expression in parathyroid glands of patients with hyperparathyroidism

Altogether 107 patients were operated on at the Department of Transplantation and Surgery of Semmelweis University in the past four years, for clinical symptoms of hyperparathyroidism. Clinical and laboratory data of the patients supported the diagnosis of primary or secondary hyperparathyroidism. Chronically impaired renal function was found in 52 cases. The removed parathyroid glands showed hyperplasia in 54, adenoma in 50 and carcinoma in 3 cases. The majority of parathyroid lesions in primary hyperparathyroidism were adenomas (41 cases) and in secondary hyperparathyroidism were hyperplasias (43 cases). The ratio of oxyphil to chief cells as well as occasional mitotic and apoptotic figures were determined. The oxyphil component was present in both hyperplastic and tumorous lesions. Apoptosis and mitosis were rarely seen in hyperplasias and adenomas (under 2%), whereas in carcinomas 3% of the tumor cells were apoptotic and 4% showed mitosis. Cytoplasmic p53 positivity could be observed in 3 of the adenomas and in 2 of the hyperplasias. The carcinomas, four adenomas and 3 hyperplasias showed nuclear p53 positivity. Bcl-2 and Bax were detected in the cytoplasm of the tumor cells in the majority of adenomas and in the cells of hyperplasias. Oxyphil cells were more frequently positive than chief cells or clear cells. Colocalization of Bcl-2 and Bax was found randomly in all types of lesions. The very low incidence of carcinoma, the low mitotic and apoptotic ratio in adenomas and hyperplasias suggest a potent antiproliferative defense mechanism in the parathyroid cell population. This may also be reflected in the cytoplasmic colocalization of various gene products which regulate cell death and cell proliferation. No significant differences in the p53, Bcl-2 and Bax spectrum were found between the primary and secondary (i.e. renal failure) parathyroid alterations.     

bcl-2和p53的表达在膀胱癌预后判断中的意义

以免疫组织化学染色的方法,研究ｐ５３和ｂｃｌ－２蛋白在膀胱癌中表达,以及与病理临床表现之间的关系。方法：应用抗ｐ５３和ｂｃｌ－２的单克隆抗体,以标准的ＬＳＡＢ方法,共分析了１３１例患者的肿瘤标本。结果：ｐ５３染色阳性的肿瘤为８６例（６５．６％）,Ｇ３级阳性的比例（８４．１％）,明显高于Ｇ１－２级（５６．３％）,Ｐ＝０．００１２）;浸润型（Ｔ２－４）肿瘤阳性的比例（７５．８％）明显高于浅型肿瘤（５５     

bcl-2、bax、p53蛋白表达与外阴鳞癌细胞凋亡的相关性研究

目的　探讨bcl 2、bax、p5 3基因与外阴鳞癌细胞凋亡的关系。方法　应用原位末端标记及免疫组化S P法检测 30份外阴鳞癌石蜡包埋组织中的细胞凋亡API和bcl 2、bax、p5 3的表达情况。 结果　外阴鳞癌组织和正常外阴皮肤中均有细胞凋亡和bcl 2、bax、p5 3的表达。随着外阴鳞癌恶性程度的增高 ,API、bcl 2、bax表达降低 ;p5 3表达增高。在外阴鳞癌的部位凋亡细胞的分布与突变型 p5 3蛋白表达呈负相关 ,p5 3分别与bcl 2及bax的表达呈负相关 (P <0 .0 5 )。在正常外阴皮肤组织中 ,凋亡细胞的分布区域与表达bcl 2及bax蛋白的细胞分布区域一致 ,bcl 2与bax的表达呈正相关 (P <0 .0 5 )。结论　外阴鳞癌的发生、发展与细胞凋亡被抑制有关 ,这一过程可能主要由 p5 3基因调控 ,bcl 2可能仅在早期起作用。     

bcl-2、bax和p53在鼻咽鳞癌中的表达及其与瘤细胞凋亡的关系

目的探讨bcl-2、bax和p53在鼻咽鳞癌中的表达及其与瘤细胞凋亡指数的关系。方法用免疫组织化学SP法检测48例鼻咽鳞癌中bcl-2、bax和p53的表达,用TUNEL法检测鼻咽鳞癌细胞的凋亡指数。结果48例鼻咽鳞癌中bcl-2、bax和p53阳性率分别为85.00%(41/48)、68.00%(33/48)和77.00%(37/48)。48例鼻咽鳞癌细胞的平均凋亡指数为25.62±25.78/HPF,凋亡指数与bcl-2、bax和p53的表达无相关性。结论鼻咽鳞癌中bcl-2和bax均呈高表达且已达到相对平衡,推测它们可能并不起到介导瘤细胞凋亡的主导作用。鼻咽鳞癌中p53的过表达可能已经失去了对bcl-2和bax表达的调节进而影响细胞凋亡的功能。     

放疗前后宫颈癌细胞凋亡及Fas、p53及bcl-2的表达

目的:探讨Fas、p53及bcl-2放疗前后的变化及与细胞凋亡的关系。方法:采用免疫组织化学SP法和脱氧核糖核酸转移酶介导的缺口末端标记(TUNEL)技术,分别检测40例宫颈鳞癌患者放疗前后同一位点凋亡阳性率及Fas、p53和bcl-2的表达。结果:放疗前后凋亡阳性率有显著性差异(P<0.01);放疗后Fas与p53表达明显升高(P<0.01),而bcl-2表达明显降低(P<0.01);放疗后p53与Fas及bcl-2表达有相关性(P<0.05)。结论:Fas、p53及bcl-2对放射线诱导的宫颈癌细胞凋亡均具有重要调控作用。p53可能通过上调Fas在宫颈癌中的表达参与了放疗诱导的细胞凋亡过程。     

凋亡相关蛋白Survivin及突变型p53在膀胱移行细胞癌中的临床意义

目的：Survivin、突变型p53在膀胱移行细胞癌（BTCC）中表达的临床意义。方法：应用SP免疫组织化学法检测60例行TUR—BT术切除BTCC标本组织石蜡切片中Survivin,突变型p53表达,结合临床资料进行分析,20例正常膀胱黏膜为对照。结果：Survivin在BTCC标本中的表达阳性率为70．0％（42／60）,而正常对照组中无1例呈阳性表达;Survivin表达与BTCC的初发、复发、病理分级有显著相关（P〈0．05）,但与肿瘤数目,临床分期无关（P〉0．05）;突变型p53在BTCC标本中表达阳性率为65．0％（39／60）,与对照组阳性率35．0％（7／20）相比差异有统计学意义（P〈0．05）,与BTCC的初发、复发、病理分级,临床分期有显著相关（P〈0．05）,它与肿瘤数目无统计学意义（P〉0．05）,BTCC中Survivin和突变型p53相比差异无统计学意义（P〉0．05）。结论：Survivin在膀胱移形细胞癌中高表达提示该基因在BTCC发生发展中有着重要的作用,Sur—vivin和突变型p53对临床医生判断BTCC的预后有着一定的临床指导意义。