大肠肿瘤中抑凋亡基因bcl－2和p53蛋白产物的表达和意义

用免疫组化方法观察４５例大肠腺瘤和６１例大肠腺癌中ｂｃ１－２和ｐ５３蛋白的表达。结果显示：正常大肠粘膜中ｂｃ１－２和ｐ５３均未见表达，而大肠腺瘤及大肠腺癌阳性率均较正常组织明显增加（Ｐ＜０．０１）．大肠腺瘤中ｂｃ１－２表达位于不典型增生较重区域。大肠腺瘤ｐ５３表达随腺瘤直径增加而增加，其中≥２０ｍｍ组阳性率显著高于＜１０ｍｍ组（ｐ＜０．０５）。ｐ５３蛋白阳性率也随不典型增生程度增加而增高。ｐ５３表达与大肠腺癌分化程度及Ｄｕｋｅｓ分期有关。大肠腺瘤中ｂｃ１－２和ｐ５３蛋白的表达呈负相关。结果表明，ｂｃ１－２蛋白表达对大肠腺瘤的增殖有一定意义，ｐ５３在大肠腺瘤癌变和大肠腺癌进展中起重要作用。     

大肠癌和腺瘤中的细胞凋亡及其调控基因表达

目的通过观察结、直肠腺癌和腺瘤中的细胞凋亡及其调控基因ｐ５３、ｂｃｌ－２的表达状态，探讨细胞凋亡及其调控基因在大肠上皮恶性转化进程的不同阶段中的作用。方法利用ＤＮＡ缺口末端标记技术和ｐ５３、ｂｃｌ－２蛋白免疫组化染色，原位观察３２例大肠绒毛状腺瘤和３３例大肠乳头状腺癌中的凋亡细胞和ｐ５３、ｂｃｌ－２蛋白阳性表达细胞的密度与分布，以１５例非肿瘤大肠粘膜做为对照。结果腺瘤和腺癌中的凋亡细胞密度均显著高于非肿瘤粘膜（Ｐ＜０．０１）；腺瘤高于腺癌（Ｐ＜０．０１）。ｐ５３和ｂｃｌ－２蛋白的表达率和表达强度，腺癌和腺瘤高于非肿瘤粘膜（Ｐ＜０．０１），腺癌的表达强度也高于腺瘤（Ｐ＜０．０５）。腺瘤中ｂｃｌ－２蛋白表达阳性组的凋亡细胞密度显著低于ｂｃｌ－２阴性组（Ｐ＜０．０１）。结论细胞凋亡调控异常在大肠癌发病中可能起重要作用。ｂｃｌ－２蛋白在腺瘤和腺癌中均可抑制细胞凋亡，而突变型ｐ５３蛋白可能仅在腺癌中起抑制细胞凋亡的作用。     

胶质瘤p53、bcl-2、MDM2表达与细胞增殖和凋亡的关系

目的：研究胶质瘤中ｐ５３ 、ｂｃｌ２ 、ＭＤＭ２ 表达与细胞增殖和凋亡的关系。方法：对４８ 例胶质瘤用免疫组化法检测ｐ５３、ｂｃｌ２、ＭＤＭ２ 蛋白表达;以Ｋｉ６７ 标记指数和ＡｇＮＯＲ 计数检测其增殖活性;用ＴＵＮＥＬ 法检测细胞凋亡。结果：ｐ５３、ｂｃｌ２、ＭＤＭ２ 蛋白表达阳性率分别为４７－９ ％ 、３５－８ ％ 及１２－５ ％ ,细胞增殖活性随肿瘤ｐ５３ 、ｂｃｌ２、ＭＤＭ２ 表达水平增高而增高,细胞凋亡则相反。结论：ｐ５３、ｂｃｌ２、ＭＤＭ２ 蛋白过表达与细胞增殖失控、凋亡抑制关系密切,在胶质瘤恶性进展中起重要作用。     

人甲状腺髓样癌内凋亡细胞检测和bcl-2、p53及Ki-67的免疫组化研究

目的：检测甲状腺髓样癌内凋亡细胞、ｂｃｌ－２、ｐ５３和Ｋｉ－６７，探讨其与患者的预后关系。方法：在２１例甲状腺髓样癌内，用ＤＮＡ末端标记方法检测凋亡细胞和用免疫组织化学技术检测ｂｃｌ－２、ｐ５３及Ｋｉ－６７的表达。结果：在２１例甲状腺髓样癌标本中，凋亡细胞ｂｃｌ－２、ｐ５３和Ｋｉ－６７的检出率分别为８１％，１００％，６２％和７１％。ｂｃｌ－２阳性物质位于肿瘤细胞的胞浆内，少数病例则位于膜上。ｐ５３蛋白阳性主要位于胞核内，少数病例也同时见于胞浆内。Ｋｉ－６７抗原主要位于肿瘤细胞核内。本研究结果表明，ｂｃｌ－２和肿瘤细胞凋亡之间有密切关系（Ｐ＜０．０１）。ｂｃｌ－２阳性细胞检出率较高的病例，凋亡细胞检出率则较低；反之，凋亡细胞检出率则较高。ｐ５３和Ｋｉ－６７阳性细胞检出率和细胞凋亡无相关性（Ｐ＞０．０５）。结论：ｂｃｌ－２具有抑制细胞凋亡的作用。上述各因素检测结果与患者存活率无统计学意义。ｂｃｌ－２肿瘤蛋白在甲状腺髓样癌中高表达，可作为该肿瘤的一个新的标记物，对预后的判断可能也有一定的意义     

食管癌变过程中p53、c-myc、bcl-2表达及其与细胞凋亡的关系

目的：探讨食管癌变过程中肿瘤抑制基因ｐ５３和癌基因ｃ－ｍｙｃ、ｂｃｌ－２的变化及其与细胞凋亡的关系。方法：采用免疫组化法（ＡＢＣ）检测２７９例食管粘膜活检组织中ｐ５３、ｃ－ｍｙｃ、ｂｃｌ－２的表达以及细胞凋亡的变化。结果：从食管正常上皮到基底细胞增生、间变和癌，ｐ５３、ｃ－ｍｙｃ、ｂｃｌ－２免疫阳性表达率及细胞凋亡发生率和细胞凋亡指数（ＡＩ）均呈升高趋势，而且在同一阶段病变，ｐ５３和ｃ－ｍｙｃ阳性表达时凋亡指数高于其阴性表达，而ｂｃｌ－２阳性表达时凋亡指数低于其阴性表达。结论：在食管癌变过程中可能有多种肿瘤抑制基因和癌基因参与，细胞凋亡在食管癌变过程中可能有重要的生物学意义。     

口腔粘膜白斑和鳞癌组织中p53和bcl—2蛋白的表达

目的：研究凋亡相关蛋白ｂｃｌ－２、Ｐ５３在口腔粘膜、上皮异常增生和鳞癌组织中的表达及意义。方法：采用免疫组织化学Ｓ－Ｐ检测１０例正常口腔粘膜,１０例单纯性增生上皮、３０例异常增生上皮和３３例鳞癌石蜡包埋组织中ｂｃｌ－２、、ｐ５３的表达。结果：正常青草帮上皮增生组织中未见Ｐ５３阳性表达。异常增生上皮和鳞癌中Ｐ５３阳性率 ３３．３３％和４５．４５％,与正常组和单纯增生组相比有显著怀差异。ｂｃｌ－２在正     

bcl-2、p53蛋白及PCNA表达与横纹肌肉瘤临床病理相关性研究

目的：研究横纹肌肉瘤（ＲＭＳ）中ｂｃｌ－２、ｐ５３、ＰＣＮＡ表达与其临床病理的相关性。方法：对５０例（随访４１例）横纹肌肉瘤进行免疫组化ＡＢＣ法标记。结果：ｂｃｌ－２、ｐ５３基因蛋白和ＰＣＮＡ，发现ｂｃｌ－２、ｐ５３、ＰＣＮＡ阳性表达率分别为２８％、７２％、７０％，其阳性表达与年龄、性别及不同组织类型的ＲＭＳ无关（Ｐ＞０．０５）。但与分化程度有关，ｐ５３、ＰＣＮＡ在低分化ＲＭＳ阳性率分别为８５％、９５％，显著高于高分化ＲＭＳ４２．８％和１４．３％（Ｐ＜０．０５），随访存活１年以内的ｐ５３、ＰＣＮＡ阳性率均为８６．７％，亦明显高于存活超过３年以上的阳性率３３．３％和４１．７％（Ｐ＜０．０５）。而ｂｃｌ－２在低分化ＲＭＳ阳性２０％显著低于高分化７１．４％（Ｐ＜０．０５），随访存活１年以内的阳性率１３．４％明显低于存活超过３年以上的４１．４％（Ｐ＜０．０５）。ｐ５３与ｂｃｌ－２阳性表达呈明显负相关，ｐ５３阳性率越高，而ｂｃｌ－２阳性率越低。结论：ＰＣＮＡ、ｐ５３、ｂｃｌ－２蛋白表达能比较准确地反映ＲＭＳ的生物学特性，ｐ５３、ｂｃｌ－２可作为肿瘤预后显著相关的有效指标。     

鼻咽癌组织中bcl—2,bax和p53的表达及其与瘤细胞？…

目的;了解鼻咽癌细胞中瘤细胞ｂｃｌ－２、ｂａｘ和ｐ５３的表达及其与瘤细胞凋亡指数的关系。方法：对３８例未经治疗的鼻咽癌组织,应用免疫组化ＬＳＡＢ法检测瘤ｂｃｌ－２、ｂａｘ和ｐ５３的表达;末端标记细胞死亡检测法（ＴＵＮＥＬ）计算癌细胞的凋亡指数。结果：（１）３７例（９７．４％）中有９０％左右的瘤细胞呈ｂｃｌ－２过表达;（２）３８例（１００％）中有７０％左右的瘤细胞过表达ｂａｘ;（３）２９例（７６．３     

bcl—2,p53蛋白在肺癌中的表达

目的：探讨ｂｃｌ－２和ｐ５３蛋白的异常表达与肺癌临床病理因素之间的关系。方法：应用免疫组化方法检测了术后随访５年以上的１２９例肺癌标本中ｂｃｌ－２、ｐ５３蛋白的表达。结果：ｂｃｌ－２及ｐ５３蛋白在肺癌中的阳性表达率分别为２８．７％和５２．７％，两种蛋白表达间呈显著正相关关系。在不同组织学类型中，两种蛋白阳性表达率间无显著差异。淋巴结癌转移阳性组中ｂｃｌ－２蛋白阳性表达率显著高于淋巴结转移阴性组（Ｐ     

食管癌变过程中p53,c—myc,bcl—2表达及其与细 …

探讨食管癌变过程中肿瘤抑制基因Ｐ５３５ 基因ｃ－ｍｙｃ，ｂｃｌ－２的变化及其与细胞凋亡的关系。方法：采用免疫组化法检测２７９例食管粘膜活检组织中ｐ５３－，ｃ－ｍｙｃ，ｂｃｌ－２的表达以及细胞凋亡的关系。结果：从食管正常上皮到基底细胞增生，间变和部，ｐ５３，ｃ－ｍｙｃ－ｂｃｌ－２免疫阳怀表达率及细胞凋亡发生率和细胞凋亡指数均呈升高趋势，而且在同一阶段病变，ｐ５３和ｃ－ｍｙｃ阳性地凋亡指数高于其阴性表     

Comparative Analysis of Cells with Combined Apoptosis and Proliferation Markers in Thyroid Tissue Specimens from Patients with Cancer,Adenoma, and Autoimmune Diseases

Combined phenotypes of cells with membrane and intracellular expression of apoptosis and proliferation regulation markers (p53, bcl-2, CD95, CD95L, Ki-67) were studied by flow cytometry of cell suspension from thyroid tissue specimens from patients with autoimmune diseases, adenoma, and thyroid cancer. The incidence of cell groups with phenotypes p53/Ki-67, p53/CD95, bcl-2/Ki-67, bcl-2/CD95, CD95/Ki-67, p53/CD95L, CD95/CD95L, and bcl-2/CD95L was evaluated and the density of receptor distribution on/in each cell group are presented. Patients with autoimmune diseases had high incidence of cells with phenotypes p53/Ki-67, p53/CD95, bcl-2/Ki-67, bcl-2/CD95, CD95/Ki-67, p53/CD95L, CD95/CD95L, and bcl-2/CD95L; cells with the bcl-2/CD95 phenotype were the most incident. Patients with thyroid adenoma had high levels of cells with p53/CD95L phenotype, while patients with thyroid cancer had significantly lower levels of p53 expression in the p53/CD95L cell group. The density of CD95L receptors on CD95/CD95L-positive cells was 4-7-fold higher in patients with thyroid tumors; the density of CD95L receptors on CD95/CD95L cells was maximum in thyroid adenoma and minimum in thyroid cancer. These data indicate differences in the expression of apoptosis and proliferation markers in thyroid adenoma, cancer, and autoimmune diseases. Analysis of the expression of these markers in the above diseases can be useful for differential diagnosis.     

Immunohistochemical analysis of bcl-2 and p53 expression in breast carcinomas: their correlation with Ki-67 growth fraction

We examined 59 breast cancers for p53 and bcl-2 protein expression by immunohistochemistry. The results were correlated with Ki-67 immunostaining. p53-negativity was noted in 40 cases and the remaining 19 tumours were p53-positive. Thirty-six tumours showed strong expression of bcl-2 and in 23 no staining for this protein was observed. We found statistically significant reverse correlation between expression of p53 and bcl-2 in majority of carcinomas: 31 cases were bcl-2 positive and p53-negative, and 14 tumours were bcl-2-negative and p53-positive. Six carcinomas showed no nuclear staining for Ki-67 and in the remaining 53 the percent of cancer cells positive for Ki-67 ranged from 1 to 60 (mean: 14.6). In these 53 cases we found that bcl-2-positive tumours were characterized by lower proliferation than bcl-2-negative tumours, the mean value of Ki-67 immunostaining being 10.7% and 23.0%, respectively. p53-negative tumours showed lower proliferation than p53-positive tumours: mean Ki-67 index was 10.2% and 23.9%, respectively.We conclude that immunohistochemically detected p53 and bcl-2 proteins show a significant inverse relationship in majority of breast carcinomas and their expression correlates with tumour proliferation (Ki-67 immunostaining).     

Proliferation and apoptosis in proliferative lesions of the colon and rectum

 Classically, neoplasia has been considered to be primarily a disturbance in the regulation of proliferation, but it is now clear that programmed cell death is dysregulated as well as proliferation. The genes that are implicated in the regulation of these processes, such as p53, c-myc and bcl-2, are often also altered in neoplasms. We have studied proliferation and programmed cell death in hyperplastic polyps, adenomas, carcinomas in adenomas and adenocarcinomas of the colorectum, using the MIB-1 antibody which recognizes the Ki-67 proliferation related antigen, and an in situ nick-end labelling procedure for histochemical labelling of proliferating and apoptotic cells. In addition, immunohistochemistry was used to study the expression of the p53, c-myc and bcl-2 proteins. The material studied consisted of 12 samples of normal mucosa, 8 hyperplastic polyps, 39 adenomas with different degrees of dysplasia and including 3 that carried a carcinoma, and 10 adenocarcinomas, all formalin fixed and paraffin embedded. The Ki-67 index indicated that proliferation increased progressively in hyperplasia, through different degrees of dysplasia in adenoma, to reach the highest level (Ki-67 index of 50%) in adenocarcinoma. Apoptosis also increased in hyperplastic polyps and in adenomas, but decreased significantly in adenocarcinomas. p53 Labelling was seen in 77% of the carcinomas but in only 3% of the adenomas. Expression of c-myc increased in adenomas and carcinomas. Furthermore, a shift from predominantly nuclear to predominantly cytoplasmic expression was seen in progressive neoplasms. Expression of bcl-2 was increased in an occasional hyperplastic polyp, but was increased markedly in almost all adenomas. Strikingly, in the adenomas with a carcinoma, the carcinoma showed weaker bcl-2 expression than the adenoma. In 20% of the carcinomas some bcl-2 staining was seen but this was less extensive than in the adenomas. Our findings indicate that in the progression from adenoma to carcinoma both increased proliferation and decreased apoptosis occur. This is paralleled by an increased expression of p53 and an increased and predominantly cytoplasmic expression of c-myc, but a decreased expression of bcl-2. This decreased bcl-2 expression does not lead to an increase in apoptotic activity. Received: 16 January 1997 / Accepted: 10 March 1997     

Proliferation in Adrenocortical Tumors: Correlation with Clinical Outcome and p53 Status

There appears to be a relationship between mitotic activity and malignant behavior in adrenocortical tumors, and carcinomas with a high mitotic index may have a poorer prognosis. This has been investigated further by quantifying and comparing the Ki-67 index using antibody MIB-1 in a series of 14 adrenocortical adenomas and 40 carcinomas. The levels have been correlated with survival and disease-free survival in carcinomas and with evidence of abnormal p53 expression as detected by immunohistochemistry. Nevertheless, many carcinomas have a low level of proliferation that may reflect varying abnormalities within the regulation of both cell division and apoptosis. Expression of bcl-2 protein, an inhibitor of apoptosis has therefore also been examined. The Ki-67 index in carcinomas was significantly higher than in adenomas, but below 4% there was overlap. There was no significant difference in survival between carcinomas with MIB-1 index <3% and those greater, but the lower group had significantly longer disease-free survival (p=0.02). There was no significant difference between p53 immunopositive and p53 immunonegative carcinomas. No tumor showed immunopositivity for bcl-2 protein. It is concluded that MIB-1 index may contribute additional prognostic information in adrenocortical tumors. Inhibition of apoptosis by bcl-2 does not appear to play a role in tumor growth.     

Immunohistochemical characteristics of duodenal adenomas in familial adenomatous polyposis with special reference to cell kinetics

The duodenum is the second most frequent site of cancer in patients with familial adenomatous polyposis (FAP). The main objective of this study was to evaluate the cell kinetics in duodenal and ampullary adenomas in FAP. The endoscopic and biopsy findings of duodenal adenomas in 22 FAP subjects and 18 non-FAP subjects were compared. Adenomas in FAP included 15 ampullary adenomas and 17 nonampullary adenomas. The cell kinetics was evaluated by immunohistochemistry for Ki-67, p53, bcl-2, and cyclooxygenase 2 (COX2), and the apoptotic index (AI) as determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method. Any correlations between the indices for cell kinetics and the endoscopic findings were identified. All 50 adenomas were histologically verified to be tubular adenoma with low-grade dysplasia. Neither the expression of Ki-67, p53, bcl-2, and COX2 nor the AI differed substantially between FAP and non-FAP subjects. In patients with FAP, duodenal adenoma tended to have a higher Ki-67-labeling index than the ampullary adenoma (54.3 +/- 11.3 versus 46.8 +/- 12.7; .05 < P < .1). In addition, the Ki-67-labeling index in endoscopically normal or slightly enlarged ampullary adenoma was significantly higher than that in markedly enlarged ampullary adenoma (51.8 +/- 11.4 versus 39.4 +/- 11.3; P < .05). Duodenal adenoma in FAP subjects was not found to have a higher proliferative activity or a smaller degree of apoptosis compared with those in non-FAP subjects. The smaller proliferative activity in larger ampullary adenoma may thus be related to the static nature of ampullary adenoma in FAP.     

Ki-67 and p53 expression in gastric dysplasias and cancer

The authors analyzed the heterogenicity of a cell population in dysplasia of varying degrees and gastric cancer from the expression of the proliferation marker Ki-67 and apoptosis activator p53 gene. Twenty-four stomachs (surgically removed for cancer) and biopsy materials (4 cases of moderate dysplasia, 4 cases of severe dysplasia) were studied. Four samples from clinically healthy individuals served as a control. The labeling index of Ki-67 in third-degree dysplasia was 37.9 +/- 2.5% and that in cancer 61.6 +/- 3.8%. There was no expression of the p53 gene in the controls. In three-degree dysplasia and cancer, the expression was 17.7 +/- 5.7% and as high as 69%, respectively. The densitometric characteristics of the labeled cells were of great diagnostic importance. Gastric cancer was represented mainly by 2 cell populations: (1) a stem cell line located in G1 with a low labeling index and a low optical density of Ki-67 and p53 and (2) an aggressive cell clone located in the S and G2 phase with marker hyperexpression.     

The Incidence of Apoptosis During Colorectal Tumorigenesis

Epithelial homeostasis in colorectal tumorigenesis is dependent not only on the rate of cell production but also on the rate of apoptosis, a genetically programmed process of autonomous cell death. Ideally, an analysis of cell kinetics should be carried out for both cell proliferation and death. We investigated the incidence of apoptosis in 35 colorectal neoplasms (15 adenomas and 20 carcinomas) using the DNA nick end labeling method (TUNEL). The expression of Ki-67 as a marker of proliferating activity and some kinds of oncogene products were analyzed immunohistochemically. When the TUNEL labeling index (TI) and the Ki-67 labeling index (KI) were determined, TI was found to be significantly higher in adenomas with high-grade dysplasia (TI: 2.5%) than in adenomas with low-grade dysplasia (TI: 0.6%) or carcinomas (TI: 1.4%). In contrast, KI increased with the progression of colorectal tumorigenesis. Moreover, TI of the carcinomas was significantly higher in c-Myc-positive cases than in c-Myc-negative cases (p<0.05). The results indicate that apoptosis plays an important role in the early stage of the adenoma-carcinoma sequence, permitting us to speculate that the increased tumor cell proliferation is negated by increased apoptosis at the stage of adenoma with high-grade dysplasia (or intramucosal carcinoma), while cell proliferation overwhelms cell death at the stage of invasive carcinoma. Int J Surg Pathol 8(2):123-132, 2000     

Expression of MUC1 and MUC2 mucins and relationship with cell proliferative activity in human colorectal neoplasia

Our previous studies on MUC1 and MUC2 mucin expression in various human neoplasms have found that MUC1 expression is related with a poor outcome whereas MUC2 expression is related with a favorable outcome. In the present study, we examined the alteration of MUC1 and MUC2 antigens on malignant transformation of colorectal mucosa, and also its relationship with cell proliferative activity (Ki-67 labeling index) of neoplastic epithelial cells in 200 adenomas and 58 carcinomas. In the 200 adenomas, we analyzed a total of 400 adenomatous lesions (mild dysplasia, 200 lesions; moderate dysplasia, 153 lesions; severe dysplasia, 47 lesions). MUC1 was expressed in carcinomas (24%) and adenomas with severe dysplasia (4%), but was not expressed in adenomas with mild or moderate dysplasia. MUC2 was expressed in a significantly greater number of adenomas with mild dysplasia (72%) than in adenomas with moderate dysplasia (45%) or severe dysplasia (47%), as well as in the carcinomas (38%; P < 0.0001). The Ki-67 labeling index was significantly lower in the MUC2-positive cases than in the MUC2-negative cases in the adenomas with mild dysplasia (13.6 vs 24.2%; P < 0.0001) or moderate dysplasia (25.7 vs 44.4%; P < 0.0001), and in the carcinomas (32.5 vs 48.4%; P < 0.05). In conclusion, the data from our study indicate that increased MUC1 expression and reduced MUC2 expression may be related to malignant transformation of colorectal neoplasia. We also demonstrated that decreased MUC2 expression, which is correlated with increased Ki-67 labeling, may play an important role in the progression of colorectal adenomatous change.