维甲酸受体-β等肿瘤分子标志物在食管正常及异常组织中的表达及意义

目的 研究食管癌高发区人群的正常食管上皮、癌前病变及癌组织活检标本中,维甲酸受体-β(RAR-β)mRNA、p16、p53和Ki67蛋白的异常表达及其与食管癌发生的关系。方法 全组397例标本,其中食管正常黏膜组织25例,轻度不典型增生69例,中度不典型增生106例,重度不典型增生51例,原位癌78例,鳞状细胞癌68例。应用RNA原位杂交技术检测RAP-β mRNA的水平,应用免疫组织化学技术检测p16、p53和Ki67蛋白的表达情况。结果 在正常黏膜、轻度、中度、重度不典型增生、原位癌及鳞状细胞癌组织中,RAR-β mRNA表达检出率分别为96．0%、89．9%、67．9％、68．6％、62．8％和63．2％;p16表达检出率分别为88．0％、71．0％、64．2％、51．0%、53．8％和52．9%;p53异常表达检出率分别为4．0％、39．1％、57．5%、52．9％、67．9％和69．1％;Ki67异常表达检出率为0、40．6％、61．3％、58．8%、59．0％和75．0%。结论 4种肿瘤生物学标志物在食管上皮中、重度不典型增生和原位癌组织中的表达,未出现明显差异,而只表现为形态学上的差异。     

食管鳞癌及其癌前病变组织p63与Ki-67蛋白共表达临床意义的探讨

目的:研究食管癌高发区食管鳞癌组织及癌前病变标本中p63和Ki-67蛋白的联合表达,探讨其共表达与食管癌变的关系及作为早期癌变生物学标志的可能性.方法:应用免疫组化技术对来自食管癌高发区的正常食管黏膜组织和轻度、中度、重度不典型增生上皮以及癌组织共203例组织中p63和Ki-67蛋白的异常表达进行研究.结果:在正常黏膜、轻度、中度、重度不典型增生及癌组织中,p63蛋白异常表达的阳性率分别为15.0%(3/20)、36.8%(14/38)、47.1%(24/51)、65.2%(30/46)和83.3%(40/48);Ki-67异常表达检出率分别为10.0%(2/20)、28.9%(11/38)、41.2%(21/51)、56.5%(26/46)和81.3%(39/48),等级相关分析结果显示,p63、Ki-67表达异常与组织学分级均显著相关(r分别为0.553和0.583,P<0.01),而且p63蛋白与Ki-67蛋白表达之间也具有相关性(r=0.690,P<0.01).结论:p63和Ki-67蛋白共表达与食管癌癌变过程显著相关,p63及Ki-67表达改变的时相分布,有可能成为在食管癌前人群中确立高危个体和选择重点化学预防个体的分子生物学标志.     

p53及PCNA的异常表达在食管上皮增生和癌变过程中的意义

目的:研究食管上皮增生、不典型增生及原位癌中p53蛋白及增殖细胞核抗原(PCNA)的异常表达,探讨其在食管癌发生发展中的作用.方法:用免疫组化LSAB方法检测189例食管鳞癌癌旁上皮及原位癌中p53及PCNA的表达.结果:癌旁上皮中存在p53蛋白积聚,从上皮增生→不典型增生→原位癌,其阳性率依次为55 %、79 %和98 %(P＜0.01).PCNA的阳性强度也是依次递增.结论:p53蛋白的积聚在食管鳞癌癌前病变中既已存在,说明它是一个早期事件,在食管癌的发生中起一定的作用.p53及PCNA的异常表达可能成为判断食管上皮发生癌变或癌前病变的客观指标之一.     

食管癌及癌前病变中p53、CD44v6蛋白的表达及其意义

目的 定位分析p53、CD44v6蛋白在食管正常黏膜、单纯增生、不典型增生、原位癌及浸润癌中的表达及与食管癌病理特征的关系。方法 采用免疫组化方法检测p53蛋白、CD44v6蛋白在45例原发性食管鳞癌大体标本中的不同病变区域的表达。结果 p63蛋白在食管癌自然病史中呈现渐进性高表达，而CD44v6蛋白则在其中呈现渐进性低表达。p53蛋白表达在单纯增生和不典型增生中有显著性差异(P＜0．01)，CD44v6蛋白则在原位癌与浸润癌中存在显著性差异（P＜0．01)。p53蛋白在食管癌自然病史中的渐进性高表达与CD44v6蛋白在其中的渐进性低表达有相关性(P＜0．01)。结论p53蛋白表达和CD44v6蛋白表达的改变与食管癌自然病史密切相关，可以作为判断食管癌生物学行为的指标。     

MDM2、P53和P27蛋白在食管鳞癌及其癌旁组织中的表达及其意义

背景与目的:探讨癌基因蛋白MDM2、抑癌基因蛋白P53以及细胞周期蛋白P27在食管鳞癌及其癌旁组织中的表达及其意义.材料与方法:采用免疫组织化学EnVision二步法(定性)检测85例食管癌存档蜡块及其癌旁黏膜中MDM2、P53和P27蛋白的表达;采用流式细胞仪(定量)检测上述3种蛋白在48例食管癌新鲜组织标本及其癌旁黏膜以及12例切缘相对正常黏膜中的表达.结果:从单纯性增生-轻度非典型增生-中度非典型增生-重度非典型增生-原位癌-浸润癌进展过程的变化,发现:定性和定量检测结果均显示P53蛋白在正常食管黏膜上皮中无表达,在食管癌变早期即出现P53蛋白的积聚;而MDM2、P27蛋白在正常黏膜上皮均有不同程度的表达,在癌变的晚期MDM2蛋白表达明显增加.结论:P53和P27蛋白表达的变化可能发生在食管癌形成早期,MDM2蛋白表达的变化可能发生在食管癌变的晚期.定性和定量2种方法联合检测能更客观和准确的探讨癌基因产物的表达及其临床意义.     

食管癌及癌前病变中p53、CD44v6蛋白的表达及其意义

目的　定位分析p5 3、CD44v6蛋白在食管正常黏膜、单纯增生、不典型增生、原位癌及浸润癌中的表达及与食管癌病理特征的关系。方法　采用免疫组化方法检测p5 3蛋白、CD44v6蛋白在 45例原发性食管鳞癌大体标本中的不同病变区域的表达。结果　p5 3蛋白在食管癌自然病史中呈现渐进性高表达 ,而CD44v6蛋白则在其中呈现渐进性低表达。p5 3蛋白表达在单纯增生和不典型增生中有显著性差异 (P <0 0 1) ,CD44v6蛋白则在原位癌与浸润癌中存在显著性差异 (P <0 0 1)。p5 3蛋白在食管癌自然病史中的渐进性高表达与CD44v6蛋白在其中的渐进性低表达有相关性 (P <0 0 1) 结论　p5 3蛋白表达和CD44v6蛋白表达的改变与食管癌自然病史密切相关 ,可以作为判断食管癌生物学行为的指标     

食管癌及癌前病变中p73、p53、ER和PR的表达及临床价值

目的:检测p73、p53、ER和PR在食管正常粘膜、单纯性增生、非典型增生及食管癌的表达及其相关性和临床病理意义,为食管癌早期诊断、预后判断、临床治疗提供有意义的生物学指标。方法:收集40例原发性食管鳞癌,14例非典型增生,14例单纯性增生和14例正常组织,采用免疫组化(S-P)方法检测p73、p53基因蛋白、雌激素受体(ER)和孕激素受体(PR)的表达。结果:p73、p53、ER和PR在食管癌、非典型增生、单纯性增生及正常组织中表达率分别为55·0%、21·4%、0%、0%;67·5%、35·7%、7·0%、0%;55·0%、21·4%、14·3%、0%;57·5%、14·3%、0%、0%。p73组从癌到正常组织的不同阶段的表达,整组比较经确切概率检验有显著性差异(P<0·001),分组比较在食管癌与非典型增生中经χ2检验有统计学意义(P<0·05),p53、ER和PR组的结果与此相似,p73与ER、PR在食管鳞癌的阳性表达具有相关性(P<0·05),与年龄、性别、肿瘤分化程度、有无淋巴结转移和浸润深度无相关性(P>0·05);ER阳性在年龄大于60岁与小于60岁比较有显著性差异(P<0·05),与有无淋巴结转移比较有显著性差异(P<0·05)。结论:p73在食管癌高表达,在正常组织与单纯组织增生无表达的特性,提示有可能成为一个新的肿瘤诊断的标记物;p73和ER、PR阳性表达在食管癌中存在相关性;p73和ER、PR联合检测可能有助于食管癌的早期诊断,判断预后,指导治疗。     

食管鳞癌组织中Cerb-B-2、P27的表达及意义

目的 探讨Cerb-B-2、P27基因的表达与食管癌发生、发展及浸润、转移的关系.方法 采用免疫组织化学S-P法检测60例食管鳞癌组织、32例癌旁不典型增生组织及60例正常食管黏膜组织中Cerb-B-2及P27蛋白表达.结果 食管鳞癌组织中Cerb-B-2蛋白表达与癌的组织学分级、浸润深度及淋巴结转移密切相关(P＜0.05);P27蛋白表达与淋巴结转移密切相关(P＜0.05);在食管鳞癌癌变过程中Cerb-B-2蛋白表达在正常黏膜组织、癌旁不典型增生组织及癌组织中的表达率依次增高,分别为25.0%(15/60)、71.9%(23/32)、86.7%(52/60),组间比较有明显差异(P＜0.05);而P27蛋白在正常黏膜组织、癌旁不典型增生组织及癌组织中的表达率依次降低,分别为88.3%(53/60)、62.5%(20/32)、31.7%(19/60),组间比较有明显差异(P＜0.05),Cerb-B-2与P27在食管鳞癌中的表达呈负相关.结论 Cerb-B-2、P27基因在食管黏膜上皮癌变过程中起重要作用;联合检测Cerb-B-2、P27基因蛋白可成为食管鳞癌早期诊断和判断预后的分子指标.     

食管鳞癌组织中p53与p27及mdm2蛋白的表达及其意义

目的:探讨癌基因蛋白p53、p27及mdm2在食管鳞癌组织中的表达及其意义。方法:采用免疫组织化学EnVision二步法对85例食管鳞癌及其癌旁组织进行p53、p27及mdm2蛋白表达的检测。结果:85例食管鳞癌组织中p53阳性表达率为70·6%(60/85),癌旁组织为75·0%(39/52);p27与mdm2蛋白在癌组织中的阳性率分别为72·9%(62/85)和83·5%(71/85),明显高于癌旁组织的34·6%(18/52)和53·8%(28/52),P<0·05;且从鳞状上皮非典型增生→原位癌→鳞癌的发展过程中,p27蛋白的阳性表达率逐渐上升。p27蛋白阳性表达与组织学分级呈正相关,r=0·234,P<0·05;而mdm2阳性表达与组织学分级呈负相关,r=-0·272,P<0·05;三者阳性表达率均与浸润深度及淋巴结转移无关,P>0·05;p53与p27和mdm2蛋白的表达无相关性,P>0·05。结论:检测p53、p27及mdm2蛋白在食管鳞癌的表达有助于了解食管癌的发生发展以及推断临床预后。     

P33~(ING1)和Survivin及Ki67蛋白在食管癌发生发展中的作用

目的 检测P33ING1、Survivin和Ki67蛋白在食管正常黏膜、单纯增生、不典型增生、浸润癌中的表达情况,分析这三种蛋白在食管癌不同病变区域中的表达及与食管癌病理特征的关系.方法 采用免疫组化方法 检测P33ING1、Survivin和Ki67蛋白在51例食管癌大体标本中的不同病变区域的表达.结果 P33ING1蛋白从正常黏膜、单纯增生、不典型增生增生、浸润癌呈渐进性低表达,而Survivin和Ki67蛋白则呈渐进性高表达.三种蛋白在正常黏膜中的表达与不典型增生及浸润癌中的表达差异有显著性(P<0.01);P33ING1蛋白在食管癌中的表达与癌肿浸润深度(肌层至全层)呈负相关(P<0.05),与分化程度(G1、G3)呈正相关(P<0.05 );Survivin蛋白表达与食管癌分化程度、淋巴结转移有关(P<0.05);P33ING1蛋白在浸润癌中的低表达与Survivin和Ki67蛋白高表达呈负相关(P<0.01).结论 Survivin功能的激活、Ki67增殖及P33ING1功能的下调可能共同对抗细胞凋亡,参与食管癌的发生发展.三者有可能为临床上食管癌早期诊断、相关治疗及预后研究提供新的手段.     

Alterations of p53, cyclin D1 and pRB expression in the carcinogenesis of esophageal squamous cell carcinoma

Many molecular alterations occur in esophageal carcinogenesis; however, little is known about the molecular genetic events responsible for the development of carcinoma. We investigated the expression of ki67, p53, cyclin D1 and pRB in 105 biopsy specimens using immunohistochemistry from iodine unstained lesions as indicators of carcinogenesis of the esophagus. Also, the genetic alternation of esophageal dysplasia from patients with accompanying esophageal squamous cell carcinoma (ESCC) was examined to study the evidence for field carcinogenesis in the esophagus. The expression of p53, cyclin D1 and pRB was detected in 31, 0 and 51.7% respectively of mild dysplasia; 40, 0 and 70% of moderate dysplasia; 40, 20 and 70% of severe dysplasia; and 48, 32 and 80% of carcinoma specimens. p53 expression was significantly increased in mild dysplasia, whereas cyclin D1 and pRB expression were significantly increased in carcinoma as compared to both normal epithelium and esophagitis. The ki67 LI and the rate of p53 expression were significantly higher in dysplasia with ESCC than in dysplasia without ESCC. Ki67, p53, cyclin D1 and pRB expression may be useful biomarkers for assessing the risk of developing esophageal cancer. Dysplasia observed at screening for secondary lesions has a highly malignant potential and careful follow-up studies are required.     

Topological analysis of p21WAF1/CIP1 expression in esophageal squamous dysplasia.

In the normal stratified squamous epithelium of the esophagus, only the third to the fifth layers of cells express the cyclin-dependent kinase inhibitor p21WAF1/CIP1 (p21). Using immunohistochemical staining, we examined the topological distribution of cells expressing p21, p53, Ki67, and cytokeratin 10 (CK10), a differentiation marker of esophageal squamous cell carcinoma (SCC), in 25 superficial SCCs and 72 dysplastic lesions of the esophagus. Image analysis of p21, p53, and Ki67 expression was also performed in 48 dysplastic lesions. In superficial SCCs, although Ki67- and p53-expressing cells were mainly distributed in the deep layers of tumors despite tumor differentiation, the distribution of p21 correlated with tumor differentiation. In dysplastic lesions, p53- and Ki67-coexpressing cells tended to locate in the same layers and expand in the lower layers of epithelium with the progression of dysplasia. p21-expressing cells shifted to the upper layers of the epithelium with the progression of dysplasia. However, this change was heterogeneous; in some lesions, p21-expressing cells were confined to the superficial layers of atypical cells (confined type), whereas in others, p21-overexpressing cells were scattered among atypical cells (scattered type). CK10 expression was observed in 25% of dysplastic lesions, and the frequency of CK10 expression was significantly higher in the scattered than in the confined type. Our results suggest that esophageal squamous dysplasia represents the earliest pathological process in esophageal squamous carcinogenesis. Our results also suggest that differentiation of esophageal SCC is determined at the stage of dysplasia, and that p21 plays a critical role in the differentiation process.     

食管癌发生发展过程中GST-π基因表达的研究

目的 探讨ＧＳＴ－π基因在食管癌发生过程中的作用。方法 采用４８例食管鳞癌标本建立食管癌发生发展多阶段模型，应用免疫组化和原位杂交方法对食管癌发生发展过程中ＧＳＴ－基因表达进行研究。结果在正常粘膜，单纯增生，不典型增生上皮总体及其Ⅰ、Ⅱ、Ⅲ级，原位癌和癌组织中ＧＳＴ－π蛋白表达阳性率分别为８７．５％，９５．３％，５５．９％，７３．９％，４７．４％，４１．２％、３６．４％和４５．８％．正常粘膜和单纯     

Analysis of Ki-67, p53 and Bcl-2 expression in the dysplasia-carcinoma sequence of Barrett's esophagus.

The grading of dysplasia in Barrett's esophagus has prognostic importance, however observer variation limits the reliability of simple histological analysis alone. We investigated Ki-67, p53 and Bcl-2 expression in Barrett's esophagus, in the sequence from Barrett's low-grade dysplasia to high-grade dysplasia and infiltrating adenocarcinoma. Forty-four esophagectomy specimens were utilized: 39 specimens with esophageal dysplasia and adenocarcinoma and 5 specimens with esophageal dysplasia only. This gave 83 sections (2 sections for specimens with dyplasia and carcinoma) examined from 44 patients. The sections were examined for Ki-67, p53 and Bcl-2 reactivity by immunohistochemistry. Low-grade dysplasia was present in 14 sections, high-grade dysplasia in 30 sections and carcinoma in 39 sections. Ki-67 expression occurred in 2 out of 14 (14%) sections with low-grade dysplasia, in 22 out of 30 (73%) sections with high-grade dysplasia and in 34 out of 39 (87%) sections with carcinoma (p<0.001). p53 protein expression was found in 1 of 14 (7%) sections with low-grade dysplasia, in 18 of 30 (60%) sections with high-grade dysplasia and in 33 of 39 (85%) sections with carcinoma (p<0.001). Expression of Bcl-2 was found in 11 of 14 (84%) sections with low-grade dysplasia but immunoreactivity was not seen in any section with high-grade dysplasia or Barrett's carcinoma. Our results indicate that overexpression of Ki-67, Bcl-2 protein and p53 mutations can be identified as early events during neoplastic progression in Barrett's esophagus. These data support the hypothesis that, in the progression of Barrett's metaplasia to adenocarcinoma, the balance of proliferation/apoptosis plays an important role.     

Reduced Fhit expression occurs in the early stage of esophageal tumorigenesis: no correlation with p53 expression and apoptosis

The FHIT gene, encompassing the FRA3B fragile site at chromosome 3p14.2, is a candidate tumor suppressor gene involved in multiple tumors, including esophageal carcinoma. We analyzed Fhit expression using an immunohistochemical method in invasive carcinoma, carcinoma in situ (CIS) and dysplasia, in paraffin sections of 75 esophageal squamous cell carcinomas (ESCs) to further elucidate the role of Fhit protein in esophageal carcinogenesis. In addition, we also examined whether Fhit expression correlated with p53 expression and apoptosis. Compared to adjacent normal mucosa, significant loss or reduction of Fhit expression was noted in 67 of 75 (89.3%) invasive ESCs, in 13 of 19 (68.4%) CIS lesions, and in 10 of 23 (43.5%) dysplastic lesions. There was a progressive loss or reduction of Fhit expression with progressive increases in the severity of histopathological changes (p < 0.001). However, there was no association between Fhit expression and clinicopathological findings, including tumor stage, lymph node metastasis, or overall survival. Moreover, Fhit expression was not significantly associated with p53 expression and apoptosis. These results indicate that abnormal Fhit expression is a common event in the early stage of ESC development and may occur independently of p53 expression and apoptosis mechanisms.     

细胞凋亡调节与宫颈癌发生

目的：探讨子宫颈癌发生中的影响细胞凋亡的调节因素。方法：检测对象为194例经福尔马林固定石蜡包埋的手术切除标本，包括上皮内瘤样病变（CIN）78例[其中重度非典型增生（SD）41例；原位癌（CIS）37例]早期浸润癌（MIC）35例；大细胞非角化型浸润癌（IC）40例及正常宫颈鳞状上皮（NE）41例。凋亡细胞的检出使用TDT－mediated dUTP-biotin nick end labeling(TUNEL)方法；细胞凋亡相关基因蛋白p53、bcl-2、bax的表达采用单克隆抗体免疫组化ABC染色方法；HPV16、18型E6 DNA感染使用PCR方法进行检测。结果：1）凋亡细胞标记率仅在CIN呈现有意义地进行性减少（P＜0．01），癌变以后不再继续下降；2）在SD、CIS组，p53、bcl-2及bax基因蛋白的表达与TUNEL标记率呈现有意义地相关性变化（P＜0．05）；3）HPV16、18型E6 DNA感染与凋亡细胞及相关基因蛋白表达未呈现相关性改变。结论：以上结果提示细胞凋亡的改变与宫颈癌发生的早期过程有关，p53、bcl-2、bax基因蛋白参与其调节，而HPV16、18型E6 DNA感染与其未呈现直接的相关性。     

Expression of GST-π gene in human esophageal carcinogenesis

Objective: To investigate the possible role of GST-π in esophageal carcinogenesis. Methods: GST-π expression at mRNA level was studied by in situ hybridization (ISH) and at protein level by immunohistochemistry (IHC). GST-π expression in normal epithelial cells (NC) of the esophagus, hyperplastic cells (HC), dysplastic cells (DC) from grade I to III, carcinoma in situ (CIS) and all the cells in squamous cell carcinomas (SCC) were examined in the same esophageal cancer specimens (n=48) which provided a model reflecting the process of esophageal carcinogenesis. Results: The positive rate of IHC staining was 87.5% for NC, 95.3% for HC, 55.9% for DC (grade I: 73.9%, grade II: 47.4%, grade III: 41.2%), 36.4% for CIS and 45.8% for SCC. The positive rate of GST-π mRNA expression was 81.2% for NC, 94.4% for HC, 61.9% for DC (grade I: 76.5%, grade II: 61.5%, grade III: 41.7%), 44.4% for CIS and 83.3% for grade I SCC, 30.0% for grade II SCC and 0% for grade III SCC. There was no statistically significant difference in GST-π expression at the mRNA and the protein level. Conclusion: There is a decreasing tendency of GST-π expression from dysplasia to CIS and SCC. The decrease in GST-π expression is an early event in esophageal carcinogenesis. This work was supported by a grant from the Key Project of Henan Province Science Foundation (No. 961001)     

Role of p53 gene mutations in human esophageal carcinogenesis: results from immunohistochemical and mutation analyses of carcinomas and nearby non-cancerous lesions

In order to characterize p53 alterations in esophageal cancer and to study their roles in carcinogenesis, we performed gene mutation and immunohistochemical analysis on 43 surgically resected human esophageal specimens, which contain squamous cell carcinoma (SCC) and adjacent non-cancerous lesions, from a high-incidence area of Linzhou in Henan, China. A newly developed immunohisto-selective sequencing (IHSS) method was used to enrich the p53 immunostain-positive cells for mutation analysis. p53 gene mutations were detected in 30 out of 43 (70%) SCC cases. Among 29 SCC cases that were stained positive for p53 protein, 25 (86%) were found to contain p53 mutations. In five cases of SCC with homogeneous p53 staining, the same mutation was observed in samples taken from four different positions of each tumor. In a well differentiated cancer nest, p53 mutation was detected in only the peripheral p53-positive cells. In tumor areas with heterogeneous p53 staining, either the area stained positive for p53 had an additional mutation to the negatively stained area or both areas lacked any detectable p53 mutation. In the p53-positive non-cancerous lesions adjacent to cancer, p53 mutations were detected in seven out of 16 (47%) samples with basal cell hyperplasia (BCH), eight out of 12 (67%) samples with dysplasia (DYS), and six out of seven (86%) samples with carcinoma in situ (CIS). All mutations found in lesions with DYS and CIS were the same as those in the nearby SCC. In seven cases of BCH containing mutations, only three had the same mutations as the nearby SCC. The results suggest that p53 mutation is an early event in esophageal carcinogenesis occurring in most of the DYS and CIS lesions, and cells with such mutations will progress to carcinoma, whereas the role of p53 mutations in BCH is less clear.     

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。