热休克蛋白10在大肠癌发生发展中的表达及其意义

目的了解热休克蛋白10（HSP10）在正常大肠粘膜、腺瘤和腺癌中的表达,并探讨其与大肠癌发生发展的关系.方法应用免疫组化Envision二步法,观察HSP10在正常大肠粘膜、腺瘤和腺癌中的表达,并比较其间是否存在差异.结果 HSP10在大肠腺瘤、腺癌中的阳性表达程度高于正常大肠粘膜（P〈0.001）;HSP10在不同程度不典型增生的大肠腺瘤、不同分化程度与淋巴结转移状态大肠腺癌中呈阳性至强阳性表达,统计学分析结果表明差异无显著性意义（P〉0.05）.结论提示HSP10与大肠癌的发生发展有关,有可能成为检测大肠癌的早期诊断标志物;HSP10的表达与大肠腺瘤的不典型增生程度和大肠癌的分化程度、淋巴结转移状态无关.     

核转录因子E2F-1和p27在大肠癌中的表达及意义

目的 探讨E2F-1和p27在大肠腺癌组织中的表达及意义.方法 采用免疫组织化学S-P法检测78例大肠癌、20例正常大肠黏膜和30例大肠腺瘤E2F-1和p27的表达情况.结果 E2F-1在大肠癌中的阳性率显著高于正常大肠黏膜和大肠腺瘤(P＜0.05).E2F-1表达与大肠癌的病理分化程度、淋巴结转移和临床分期具有相关性(P＜0.05).p27在正常大肠黏膜的阳性率明显高于大肠腺瘤和大肠癌.p27表达与大肠癌的病理分化程度、淋巴结转移和临床分期具有相关性(P＜0.05).结论 E2F-1和p27的表达与大肠癌发生关系密切,可以作为肠癌发生、发展的生物学标志物.     

大肠癌发生发展中自噬基因Beclin1的表达及其与增殖和凋亡相关基因的关系探讨

探讨在大肠癌发生发展过程中自噬基因Beclin1的表达及其与增殖、凋亡相关基因之间的关系。方法:采用免疫组化检测29例正常大肠黏膜、60例大肠腺瘤、19例腺瘤恶变及50例大肠癌组织中Beclin1、Bcl-2、Caspase-9、Caspase-3及Ki-67蛋白的表达。结果:1）Beclin1在大肠癌中的阳性表达率高于正常大肠黏膜,但与大肠癌的组织学分化无关,与淋巴结转移有关;2）Bcl-2在大肠腺瘤、腺瘤恶变及大肠癌组织中的表达高于正常大肠黏膜,且与大肠癌的组织学分化及转移有关;Caspase-9与大肠癌的癌变有关,但与组织学分化和淋巴结转移无关;Caspase-3在正常大肠黏膜中的阳性表达高于其他组,无淋巴结转移组高于淋巴结转移组,但与大肠癌的组织学分化无关;3）Ki-67在正常大肠黏膜、大肠腺瘤、腺瘤恶变及大肠癌中的表达逐渐增高,差异有统计学意义,且与大肠癌的组织学分化和淋巴结转移有关。结论:1）自噬可能与大肠癌的发生有关,但随着肿瘤的进展,自噬能力降低可能会促进其侵袭转移。2）大肠癌发生发展过程中,在癌细胞无限增殖的同时,自噬、凋亡通过内在的分子机制相互协调转化,使癌细胞逃脱了机体正常的清除机制,向恶性方向发展。      

表皮-钙黏附素和Snail蛋白的表达与大肠癌侵袭转移及预后的关系

目的 研究表皮-钙黏附素(E-CD)和Snail蛋白在大肠癌组织中的表达及其与大肠癌侵袭、转移和预后之间的关系.方法 采用免疫组化EnVision法,检测E-CD和Snail蛋白在30例正常大肠黏膜、30例大肠腺瘤及142例大肠癌组织中的表达.结果 E-CD蛋白在正常大肠黏膜组织中的强阳性表达率为90.0%,明显高于其在大肠腺瘤和大肠癌组织中的强阳性表达率(63.3%和41.5%,P<0.05).Snail蛋白在大肠癌组织中的阳性表达率为52.1%,明显高于其在正常大肠黏膜和大肠腺瘤组织中的阳性表达率(6.7%和26.7%,P<0.05).E-CD蛋白的表达与大肠癌的分化程度、浸润深度、静脉侵犯、淋巴管侵犯、淋巴结转移以及Duke's分期有关(均P<0.05),而与患者的年龄、性别、肿瘤大小以及病理组织学类型无关(均P0.05).Snail蛋白的表达与大肠癌的分化程度、浸润深度、静脉侵犯、淋巴管侵犯、淋巴结转移、病理组织学类型以及Duke's分期有关(均P<0.05),而与患者的年龄、性别以及肿瘤大小无关(均P>0.05).E-CD蛋白与Snail蛋自在大肠癌组织中的表达呈负相关(r=-0.508).E-CD蛋白阳性表达者的术后1、3和5年生存率明显高于阴性表达者(P<0.05),Snail蛋白阴性表达者的术后1、3和5年生存率明显高于阳性表达者(P<0.05).淋巴结转移、Duke's分期、E-CD和Snail蛋白的表达可作为大肠癌独立的预后指标(均P<0.05).结论 E-CD和Snail蛋白的表达与大肠癌的侵袭、转移和预后明显相关,E-CD蛋白表达降低和Snail蛋白表达增强的大肠癌患者病期晚、预后差.     

良恶性大肠肿瘤组织血管内皮生长因子表达与意义

目的:研究血管内皮生长因子(VEGF)在大肠癌、大肠腺瘤和癌旁正常大肠黏膜标本中的表达,并探讨其表达与大肠癌发生、发展及转移的关系.方法:通过免疫组化SP法检测89例大肠癌、22倒大肠腺瘤、26例癌旁正常大肠黏膜标本中VEGF表达及与大肠癌临床病理特征的关系.结果:VEGF在大肠癌组织中的阳性率(71.9%,64/89)明显高于其在癌旁正常大肠黏膜组织(30.8%,8/26)、大肠腺瘤组织(40.9%,9/22)中的阳性率,差异有统计学意义(P<0.01);VEGF在大肠癌组织中的表达水平与肿瘤Dukes分期,有无淋巴结转移及患者年龄有关(P<0.05),与肿瘤组织学分级和患者性别无关.结论:VEGF表达升高与大肠癌的发生、发展、浸润及淋巴结转移有关,检测VEGF的表达情况有助于判断大肠癌的恶性程度及肿瘤预后.     

HSP27和GST在大肠腺瘤和大肠癌中表达的临床病理研究

目的探讨热休克蛋白27（HSP27）和谷胱甘肽-S-转移酶（GST）在正常大肠黏膜、大肠腺瘤和大肠癌中的表达及其临床病理意义。方法应用免疫组织化学S-P法检测HSP27和GST在大肠组织中的表达。结果HSP27在正常肠黏膜、大肠腺瘤、无淋巴结转移的大肠癌、有淋巴结转移的大肠癌中表达阳性率分别为30％,33．3％,65％,70％,多组间比较具有显著性差异（x^2=12．723,P=0．013）。两组间比较发现,HSP27表达率在正常组与大肠癌组存在差异,在有淋巴结转移的大肠癌组中的表达具有差异最为显著（x^2=8．688,P=0．003）,而其它组间比较无显著性差异。GST在正常肠黏膜、大肠腺瘤、无淋巴结转移的大肠癌、有淋巴结转移的大肠癌中表达阳性率分别为60％,80．9％,90％,100％,多组间比较具有显著性差异（x^2=16．707,P=0．001）;进一步进行两组间比较发现,GST表达率在有淋巴结转移的大肠癌组中的表达具有差异最为显著（x^2=10．909,P=0．001）,而其它组间比较无显著性差异。GST和HSP27表达无显著相关。结论HSP27和GST在大肠癌中的表达可能在大肠癌发生发展中发挥作用,特别在大肠癌恶性演进中以及预后预测中具有重要意义,可能作为独立的生物标志物。     

粘蛋白MUC1、MUC2在大肠腺癌中的表达及其临床意义

 目的　探讨粘蛋白MUC1和MUC2在大肠腺癌及大肠腺瘤中的表达及其与临床各个病理参数之间的关系。方法　应用免疫组织化学方法对 6 0例大肠腺癌和 2 0例大肠腺瘤进行粘蛋白MUC1、MUC2检测。结果　 2 0例大肠腺瘤及 6 0例大肠腺癌中 ,粘蛋白MUC1阳性表达率分别为 10 %、4 6 .7% ;粘蛋白MUC2阳性表达率分别为 10 0 %、5 8.3%。大肠腺癌中粘蛋白MUC1的表达与肿瘤的分化程度呈负相关 ,与浸润深度、淋巴结转移、Dukes分期及生存期呈正相关。MUC2的表达与分化程度无关 ,而与浸润深度、淋巴结转移、Dukes分期及生存期均呈负相关。结论　MUC1的上调表达或MUC2的下调表达可能参与了大肠腺癌的发生、发展、浸润及转移 ,对临床上判断预后具有较大的意义。     

大肠肿瘤中Ezrin的表达及意义

目的:检测大肠癌及大肠腺瘤中Ezrin蛋白的表达,了解与大肠癌临床病理的相关因素。方法:采用SP免疫组化法,检测Ezrin蛋白在各组织中的表达及意义。结果:Ezrin在大肠癌、大肠腺瘤中的阳性表达率分别为69.4%(43/62)和40.0%(4/10),癌旁组织阳性表达率为32.6%(15/46),正常大肠组织阳性表达率为6.7%(1/15)。Ezrin的表达在正常大肠组织和大肠癌之间、大肠腺瘤和大肠癌之间以及癌旁组织和大肠癌之间均有显著性差异(P<0.05),而癌旁组织和大肠腺瘤之间Ezrin表达差异无显著性(P>0.05);Ezrin在大肠癌转移组及未转移组中的阳性表达率分别为89.7%(26/29)和51.5%(17/33);转移组阳性表达率高于未转移组(P<0.05);大肠癌组织中Ezrin的表达与患者年龄、性别、肿瘤发生部位、分化程度均无关(P>0.05)。结论:Ezrin的表达增高与大肠癌转移密切相关,可作为临床判断大肠癌转移及预后等生物学行为的重要参考指标。     

人大肠腺瘤和腺癌之间相关差异蛋白的蛋白质组学研究

目的　应用蛋白质组学的方法,建立人大肠腺癌组织、大肠腺瘤组织、正常大肠黏膜组织的二维电泳图谱,筛选并鉴定三者之间的差异蛋白,发现人大肠癌的潜在生物学标志物。方法　收集同时患有大肠腺瘤的大肠腺癌患者１０例,术前及术后病理证实均为Ｄｕｋｅｓ’Ｂ期。取同一患者术后大肠癌组织、距大肠癌组织１５ｃｍ以上正常大肠黏膜组织以及肠镜下切除大肠腺瘤组织,提取不同组织总蛋白进行二维电泳,用ＩｍａｇｅＭａｓｔｅｒ２ＤＰｌａｔｉｎｕｍ５.０凝胶图像分析软件对双向电泳图依次进行凝胶点的检测、背景消减和编号。识别不同组织之间的差异蛋白点,对差异蛋白点用胶内酶解后行质谱分析和网上数据库鉴定。结果　大肠腺癌组织、大肠腺瘤组织和正常大肠黏膜组织之间存在９０个差异蛋白点。对这些差异点进行质谱分析,经数据库鉴定出７１个蛋白质。经分析,大肠腺瘤组织与正常大肠黏膜组织相比,１７个蛋白表达上调,９个蛋白表达下调;大肠腺癌组织与正常大肠黏膜组织比较,４７个蛋白表达上调,１２个蛋白表达下调;大肠腺癌组织与大肠腺瘤组织相比,３５个蛋白表达上调,８个蛋白表达下调。结论　大肠腺癌组织、大肠腺瘤组织和正常大肠黏膜组织之间存在差异蛋白表达,可能与大肠腺癌的发生发展有关;应用蛋白质组学方法筛选人大肠腺癌发生发展相关蛋白是可行的。     

自噬相关基因Beclin-1与抑癌基因PTEN、p53在大肠癌中的表达和意义

目的:检测自噬相关基因Beclin-1与抑癌基因PTEN、p53在大肠正常黏膜、大肠腺瘤、大肠癌组织中的表达,探讨它们与大肠癌发生发展的相关性。方法:采用免疫组织化学SP法检测Beclin-1与PTEN、p53在15例大肠正常黏膜、30例腺瘤及65例大肠癌组织中的表达,结合临床病理因素进行分析。结果:Beclin-1在大肠癌组阳性表达率高于正常大肠黏膜组(P<0.001)与大肠癌组织学分化程度、Dukes分期及淋巴结转移无关。PTEN在大肠正常黏膜组中的表达高于大肠癌组(P=0.001),与大肠癌组织学分化程度、Dukes分期及淋巴结转移有关。p53在大肠正常黏膜、腺瘤及大肠癌组织中的表达逐渐增高(P<0.001)与大肠癌组织学分化程度、Dukes分期及淋巴结转移有关。经Spearman秩相关检验,Beclin-1蛋白与PTEN蛋白相关系数为-0.289,呈负相关表达;Beclin-1蛋白与p53蛋白相关系数为0.347,呈正相关表达。结论:Beclin-1、PTEN、p53在大肠癌中的异常表达与大肠癌的发生、发展密切相关。自噬可能与大肠癌的发生相关,自噬活性的上调与大肠癌发生有关。自噬相关基因Beclin-1与抑癌基因PTEN、p53在大肠癌的发生发展中起协调作用。同时检测对判断大肠癌的进展程度具有重要的意义。     

热休克蛋白10在大肠癌的表达及其临床病理意义

Objective： To investigate the expression of heat shock protein 10 （HSPIO） during genesis and development of large bowel carcinoma and discuss the clinical significance about its expression. Methods： The expression of HSPIO was observed in specimens from normal colonic mucosa （NC）, colorectal adenomas （CA） and colorectal adenocarcinomas （CAC） by immunohistochemistry EnVisionTM. Its correlations to clinicopathologic features, as well as to postoperative survival time of large bowel carcinoma patients were analyzed. Results： The expression of HSPIO was common in normal colonic mucosa, colorectal adenomas and adenocarcinomas and more intensive in colorectal adenomas and adenocarcinomas than that in normal colonic mucosa （P 〈 0.001）. The positive expression of HSPIO had no correlation to clinicopathologic features, including age, gender, primary tumor, infiltrating of regional lymph node, metastasis, clinical stage and histopathology of large bowel carcinoma patients, as well as to their postoperative survival time. Conclusion： HSPIO was overexpressed in the early stage of colorectal adenocarcinoma suggesting that it could serve as an index for early diagnosis of large bowl carcinoma. The positive expression of HSPIO had no correlation to clinicopathologic features or postoperative survival time of large bowel carcinoma patients.     

Karyometry of the colonic mucosa.

OBJECTIVE: The study summarizes results of karyometric measurements in epithelial cells of the colorectal mucosa to document evidence of a field effect of preneoplastic development among patients with colorectal adenocarcinoma or adenoma. METHODS: Karyometric analyses were done on high-resolution images of histologic sections from 48 patients with colorectal adenocarcinomas and 44 patients with adenomas and on images from matching normal-appearing mucosa directly adjacent to such lesions, at a 1-cm and 10-cm distance from the lesions or from the rectal mucosa of adenoma patients, as well as from 24 healthy normal controls with no family history of colonic disease. RESULTS: The nuclei recorded in the histologically normal-appearing mucosa of patients with either colorectal adenoma or adenocarcinoma exhibited differences in karyometric features in comparison with nuclei recorded in rectal mucosa from patients who were free of a colonic lesion. These differences were expressed to the same extent in tissue adjacent to the lesions and in normal-appearing tissue as distant as the rectum. CONCLUSIONS: The nuclear chromatin pattern may serve as an integrating biomarker for a preneoplastic development. The field effect might provide an end point in chemopreventive intervention trials.     

大肠肿瘤中bcl—2及PCNA表达及其意义

目的：研究ｂｃｌ－２及ＰＣＮＡ在大肠肿瘤肆生中的作用及其临床意义。方法：应用免疫组化Ｓ－Ｐ法分别检测大肠正常粘膜、腺瘤及腺癌中ｂｃ；－２及ＰＣＮＡ表达。结果：Ｂｃｌ－２在正常粘膜基底部上皮细胞表达，在腺瘤（７７．５％）和腺癌中（５６．３％）ｂｃｌ－２表达差异显（Ｐ〈０．０５）。高分化腺癌ｂｃｌ－２表达率（６８．４％）显高于差分化腺癌（４１．７％），ＰＣＮＡ表达在正常粘膜，腺瘤及腺癌中依次递增，     

Detection of the tumor-associated glycoprotein antigen (TAG-72) in premalignant lesions of the colon

We used monoclonal antibody B72.3 to study the expression of the colorectal carcinoma-associated antigen TAG-72 in premalignant colonic lesions with the immunoperoxidase technique. This antigen, which is rarely detectable in the normal colonic epithelium, was expressed in 13 of 19 adenomas with moderate to severe dysplasia and nine of nine cases of inflammatory bowel disease. The antibody reacted with the normal-appearing mucosa adjacent to a carcinoma in 10 of 12 cases, although only eight of the tumors expressed the antigen. The expression of the TAG-72 antigen in the colonic epithelium may be an early marker of malignant transformation.     

Colorectal ornithine decarboxylase activity in human mucosa and tumors: elevation of enzymatic activity in distal mucosa

To examine the value of ornithine decarboxylase (ODC) assay as a biological marker of potential malignancy in large bowel, we harvested 43 colorectal carcinoma, 7 adenoma, 6 polyps, and 77 normal-appearing mucosa at surgery from patients with colorectal carcinoma. In addition, 13 normal rectal mucosa were obtained at biopsy from patients with benign diseases or diseases unrelated to colorectal carcinoma as normal control. ODC activity was significantly higher in polyps and adenocarcinomas than in normal-appearing mucosa from patients with colorectal carcinoma. ODC activity in normal-appearing mucosa varied throughout the large intestine, with significantly higher activities in the distal segment of the large bowel. The higher ODC activity detected in the sigmoid colon and rectum correlates with the larger incidence of tumor development in this region of the large bowel. This observation needs to be taken into consideration when ODC activities of the colorectal mucosa are measured as biological markers of potential malignancies.     

Prognostic and diagnostic significance of beta-catenin nuclear immunostaining in colorectal cancer.

In the present study, we investigated the prognostic and diagnostic significance of beta-catenin nuclear immunostaining in 60 specimens of normal colorectal tissue; 180 specimens of colorectal polyps, adenomas, and carcinomas; and 40 specimens from patients with the simultaneous occurrence of polyps, adenomas, and carcinomas. Additional specimens from 59 patients with colorectal carcinoma and 14 patients with adenoma who subsequently developed carcinoma were examined for possible survival study. Immunohistochemical staining showed that the occurrence of nuclear beta-catenin correlated with the sequential stages in colorectal carcinogenesis, in which positive staining was observed in 0% of normal tissues, 8% of polyps, 92% of adenomas, and 100% of carcinomas. High immunohistochemical scores in colorectal carcinoma were significantly associated with lymph node metastasis and poor survival. Adenomas associated with synchronous or metachronous carcinomas showed significantly higher levels of nuclear beta-catenin compared with adenomas without associated carcinomas. Nuclear translocation of beta-catenin was rare or absent in other types of cytokeratin 20 positive adenocarcinomas examined (99 cases). Thus, it was positive in only 7% of colonic mucinous adenocarcinomas, 3% of pancreatic adenocarcinomas, 8% of ovarian mucinous cystadenocarcinomas, and 0% of gastric adenocarcinomas. However, 100% of primary and metastatic colorectal adenocarcinomas were positive for nuclear staining for beta-catenin. Thus, nuclear staining for beta-catenin may serve as an additional parameter to help distinguish colorectal adenocarcinomas from adenocarcinomas of other tissue sites. Collectively, the present large-scale study has clearly addressed the clinical significance of beta-catenin nuclear translocation with respect to tumor progression, survival, and differential diagnosis.     

PROLIFERATION RATE OF COLONIC NEOPLASMS AND MUCOSA ADJACENT TO NEOPLASMS: IMMUNOHISTOCHEMICAL STUDY

Biopsy specimens of normal mucosa (n=5). adenomas (n -13), adenocarcinomas (n = 8). mucosa adjacent to adenoma (n=10) mucosa adjacent to adenocarcinoma (n = 7 ) at the large bowel were investigated by an iminunohistochemical method using 5- bromodeoxynldine (BrdUrd) . The labeling index (LI) was significantly lower in normal nucosa than mucosa adjacent to neoplasms and adenomas and adenocarcinomas. The proliferative zone was confined to the lower two- third at the crypt in normal mucosa and in mucosa adjacent to neoplasms. The labeled cells were either present in the upper third or scattered along the crust and in surface epithelium. The results support the adenmo-carcinoma sequence.     

Abnormalities in the expression of cell cycle-related proteins in tumors of the small bowel.

Tumors of the small bowel are quite rare for unknown reasons, although they resemble colorectal tumors in many respects. The purpose of this study was to determine whether abnormalities in the expression of several cell cycle control genes are of importance in small bowel tumorigenesis by comparing a series of samples of normal mucosa, adenomatous polyps, and adenocarcinomas. The levels of cyclin D1, cyclin E, p16, p21, p27, and p53 proteins were determined by immunohistochemistry in samples of normal small bowel (n = 16), small bowel adenomas (n = 20), and small bowel adenocarcinomas (n = 24). Normal small bowel mucosa expressed p27 protein, but not the other cell cycle-related proteins. About 20% of the tumors displayed a decrease in the expression of this protein. The most frequent alteration in the tumors was an increase in the p16 protein. Increased expression of p53 was associated with tumor progression because it was overexpressed in 45% of the adenomas and 65% of the adenocarcinomas (P<0.05). Advanced age and increased detection of cyclin D1 and p53 were associated with a decreased 3-year survival (P<0.05). Cell cycle abnormalities are early and important events in the multistep process of small bowel tumorigenesis, thus resembling colorectal carcinogenesis. As in colon cancer, deregulated expression of G1 proteins may perturb cell cycle control in benign adenomas of the small bowel and thereby enhance tumor progression. Increased expression of cell cycle inhibitors in tumors may serve as a defense mechanism for tumor progression.     

Evidence for the existence of different types of large bowel tumor: suggestions from the clinical data of a population-based registry

The clinical findings of a population-based colorectal tumor registry have been analyzed to determine elements of supporting or not supporting the existence of different types of large bowel cancer. Age-specific incidence rate of the 409 registered patients rose sharply with increasing age in all segments of the large bowel; however, regarding left colon and rectum, the male: female ratio showed a marked male preponderance, more evident in the more advanced age groups. Histopathology, studied in 87% of patients, revealed adenocarcinoma as the most frequent feature; however, adenocarcinoma with concomitant adenoma (i.e., presumably arising in adenoma) was observed in 14.3% of cancers of the left colon, in 17.7% of rectal tumors, but in only 5.7% of neoplasms of the proximal colon (P less than 0.05 and P less than 0.01, respectively, vs. left colon and rectum). Some histological features (carcinoid and mucinous carcinoma) were observed in right-side tumors only. Analysis of the familial occurrence of cancer showed that a significantly larger proportion of patients with neoplasms located in proximal colonic segments had three or more first-degree relatives affected by (or deceased from) cancer of all sites. Similarly, colorectal tumors among relatives were more frequent in patients with right-side cancer. The location of the 793 polyps observed during 3 years of registration showed that more than 70% of adenomas were located beyond the splenic flexure, overlapping the distribution of cancers. In conclusion, the differences of sex ratio at different colonic subsites, the higher fraction of adenocarcinomas with adenomas in cancer of the more distal tracts of the large bowel, and the more marked familial occurrence of colorectal cancer in patients with right-side neoplasms tend to support the view that cancer of the proximal colon, cancer of the distal colon, and cancer of the rectum may actually be three different types of tumors.