溃疡性结肠炎相关结直肠癌临床特点及癌变相关蛋白的表达

目的 探讨溃疡性结肠炎(UC)相关结直肠癌的临床特点及其癌变的可能机制.方法 收集北京协和医院1984年至2008年6例UC相关性结直肠癌,分析其发病情况、临床表现、病理类型、治疗及预后特点,免疫组化检测组织标本中结肠腺瘤性息肉蛋白(APC蛋白)、β-连环蛋白、p53蛋白和Wnt1蛋白表达的情况.结果 UC的癌变率为1.1%(6/534),女性多见(5/6),平均病程14.3年.临床均呈现典型的UC表现,病变常累及全结肠(5/6),均无原发性硬化性胆管炎.其中直肠癌4例、降结肠癌2例,病理类型以腺癌为主,预后较差.APC蛋白、β-连环蛋白、p53蛋白和wnt1蛋白表达的阳性率分别为6/6、6/6、5/6和6/6.结论 临床对病变累及全结肠、病程长的UC患者,应注意防止结直肠癌的发生,其癌变过程可能有多途径参与.     

p73和p51基因在结直肠癌中的表达和意义

背景：p53基因是一种肿瘤抑制基因，其家族成员p73和p51在结构上与p53具有高度同源性，影响细胞转录和凋亡的功能与p53相似。目的：研究p73和p51基因在结直肠癌中的表达及其与细胞凋亡和肿瘤临床病理特征的关系，探讨两者在结直肠癌发生、发展中的可能作用。方法：以逆转录聚合酶链反应（RT—PCR）检测60例结直肠癌组织和相应癌旁组织中p73、p51mRNA表达，以原位末端标记（TUNEL）法检测细胞凋亡。结果：结直肠癌组织p73、p51AmRNA表达阳性率显著高于相应癌旁组织（p73：71．7％对5．0％，P〈0．01：p51A：46．7％对11．7％，P〈0．01）：p51B mRNA在结直肠癌组织与相应癌旁组织中的相对表达量无明显差异（0．7318±0．3628对0．6836±0．3516，P〉0．05）。p73、p51A mRNA表达阳性者肿瘤细胞凋亡指数分别显著低于p73、p51A mRNA表达阴性者（p73：3．2％±2．5％对5．5％±2．8％．P=0．003；p51A：2．6％±2．3％对4．9％±2．7％，P=0．001）。p73mRNA表达与结直肠癌的分化程度、TNM分期和淋巴结转移相关（P〈0．05），p51A mRNA表达仅与淋巴结转移相关（P〈0．05）。结论：结直肠癌中p73、p51A基因表达上调，两者可能通过抑制肿瘤细胞凋亡而参与了结直肠癌的发生、发展。p73过表达可能与结直肠癌预后不良有关。     

c-erbB-2 、bcl-2和p53在结直肠肿瘤中的表达及其临床意义

背景：c-erbB-2、bcl-2和突变型p53在结直肠腺瘤癌变过程中相互调节并发挥重要作用。目的：探讨结直肠肿瘤中c-erbB-2、bcl-2和D53蛋白的表达及其临床意义。方法：取42例结直肠腺癌、10例腺瘤和10例正常结直肠黏膜组织，以免疫组化方法检测其中c-erbB-2、bcl-2和p53蛋白的表达，分析其表达与腺癌临床病理特征的关系。结果：c-erbB-2、bcl-2和p53蛋白在腺癌、腺瘤和正常黏膜组织中的表达差异均有统计学意义（P〈0．05），bcl-2蛋白在腺癌组织中的表达低于腺瘤组织，c-erbB-2和p53蛋白在腺癌组织中的表达高于腺瘤和正常黏膜组织。c-erbB-2蛋白的表达随腺癌分化程度的降低而增高，bcl-2蛋白的表达随腺癌分化程度的降低而降低：c-erbB-2和p53蛋白的表达与淋巴结转移和Dukes分期呈正相关。腺癌组织中bcl-2与p53蛋白的表达呈负相关（rs=-0．301，P〈0．05）。结论：c-erbB-2与结直肠癌的进展、分化和转移相关。腺癌组织中p53高表达和bcl-2相对低表达提示两者可能参与了结直肠癌的发生、发展过程，bcl-2过表达可能在结直肠癌发生的早期起作用。     

溃疡性结肠炎结肠上皮TLR4和HBD2表达的研究

目的通过检测UC结肠上皮TLR4、MD2和HBD2的转录与表达,探讨TLR4和HBD2在UC发生发展中的可能作用。方法选取60例研究对象,其中50例活动期UC设为UC组,10例IBS设为对照组,均在结肠镜下于乙状结肠处取活检行进一步检测。首先进行UC疾病活动度(UCAI)的计算和病理学分级,再免疫组化检测每例结肠上皮的TLR4和HBD2表达,RT-PCR方法测定TLR4、HBD2 mRNA水平,免疫印迹方法检测TLR4蛋白的表达。结果 UC组患者UCAI评分与病理分级呈正相关;免疫组化显示TLR4和HBD2蛋白在UC结肠上皮和炎症细胞中的表达均高于对照组,阳性率与病理学分级呈正相关,且TLR4和HBD2二者表达呈正相关;UC结肠上皮的TLR4、MD2和HBD2 mRNA水平明显高于对照组,也与病理学分级呈正相关。免疫印迹检测表明UC结肠上皮TLR4蛋白表达显著高于对照组,并与病理学分级呈正相关。结论 TLR4和HBD2在活动期UC结肠上皮表达显著上调,且二者呈正相关,这表明UC的发生发展可能与细菌入侵肠上皮进而导致其免疫失衡有关。     

结直肠癌PTEN基因表达及与ERK2、p27kip1的相关性

目的:研究结直肠癌中的PTEN、ERK2及p27kip1蛋白的表达及相互关系,初步探讨他们在结直肠癌发生发展中的生物学意义.方法:用免疫组织化学染色快捷法,检测40例结直肠癌组织、18例结直肠腺瘤、13例结直肠正常黏膜中PTEN蛋白、p27kip1和ERK2蛋白的表达,比较PTEN蛋白表达与临床病理指标的关系,及其与p27kip1、ERK2蛋白表达的相关性.结果:结直肠癌癌组织PTEN,ERK2和p27kip1蛋白表达的阳性率与腺瘤及正常组织间比较差异有显著性(57.5% vs 72.2%,100%;70.0% vs 61.1%,23.1%;62.5% vs 77.8%,100%;P＜0.05);PTEN蛋白表达强度与ERK2蛋白表达强度之间呈负相关(r=-0.452,P＜0.05),与p27蛋白表达强度呈正相关(r=0.379,P＜0.05);PTEN,p27kip1蛋白与结直肠癌分化程度、淋巴结转移及Dukes分期相关(P＜0.05);ERK2蛋白随结直肠癌淋巴结转移、Dukes分期的进展而增高.结论:抑癌基因PTEN的表达与结直肠癌生物学行为密切相关;在结直肠癌发生、发展过程中,可能由于PTEN蛋白的低表达或失表达抑制p27kip1蛋白表达及Ras/Raf/MEK/ERK信号通路的异常激活,使细胞发生癌变,并促进癌变细胞的浸润、转移.     

Beclin1和PTEN蛋白在结直肠癌中的表达及预后价值

目的 研究Beclin1和PTEN蛋白在结直肠癌组织和癌旁组织中表达的差异及其与结直肠癌预后的相关性。方法 收集2007～2017年在解放军第九四医院就诊并手术的结直肠癌患者127例的结直肠癌癌灶组织和相对应的癌旁组织及每个患者的临床资料，行免疫组化检测癌组织和癌旁组织的Beclin1、PTEN的定位，并检测两个基因的表达阳性率，用实时PCR方法检测组织中Beclin1、PTEN mRNA表达，用Western印迹检测其中Beclin1、PTEN的蛋白表达，并比较2个基因在癌组织和癌旁组织中的表达差异。对Beclin1和PTEN的蛋白表达进行相关性分析，判断两个基因之间是否存在可能的调控关系。并对患者的临床资料进行生存分析，了解Beclin1、PTEN与结直肠癌患者预后间的关系。结果 Beclin1、PTEN均主要定位于结直肠细胞的胞质中，胞核中有少量表达；Beclin1在癌灶组织中的基因和蛋白表达显著高于癌旁组织，而PTEN在癌组织中的mRNA和蛋白表达显著低于癌旁组织(P<0.01)。双变量直线分析结果显示，在癌和癌旁组织中Beclin1和PTEN的蛋白表达呈负相关(P<...     

Cdx2和MUC2在反流性食管炎、Barrett食管和食管腺癌中的表达

目的研究Cdx2和MUC2在反流性食管炎、Barrett食管和食管腺癌中表达，探讨3种食管黏膜疾病的内在关系。方法选取反流性食管炎30例、Barrett食管18例及食管腺癌25例作为研究对象，以正常食管上皮黏膜25例作为对照，采用免疫组化方法检测Cdx2和MUC2的表达，对结果进行统计分析。结果Cdx2和MUC2在反流性食管炎、Barrett食管及食管腺癌中的蛋白阳性表达率均较正常对照组明显增高（P〈0．05）。Cdx2在正常食管黏膜上皮中无表达，在反流性食管炎、Barrett食管及食管腺癌中的阳性表达率分别为26．7％、66．7％和28．0％，在Barrett食管中表达明显高于反流性食管炎（P〈0．05），亦明显高于食管腺癌（P〈0．05）；MUC2在正常食管黏膜上皮和反流性食管炎组织无表达，在Barrett食管及食管腺癌中的阳性表达率分别为61．1％和24．0％，Barrett食管中表达率明显高于食管腺癌（P〈0．05）。两者表达情况相似。结论Cdx2是肠上皮化生的始动因素，MUC2的表达是肠上皮化生的晚发事件。Cdx2和MUC2在反流性食管炎、Barrett食管和食管腺癌组织中的表达情况支持这3种食管黏膜疾病间有密切的关系。     

Nucleostemin在结直肠癌和正常结直肠组织中的表达及其意义

背景：Nucleosterain（NS）是一个与细胞增殖相关的核蛋白,部分研究显示其仅高表达于干细胞和肿瘤细胞,成体组织分化细胞中未见NS表达。目的：探讨NS在结直肠癌和正常结直肠组织中的表达及其意义。方法：收集38例结直肠癌患者的癌组织及其相应癌旁正常组织标本．以实时RT—PCR和蛋白质印迹法检测NSmRNA和蛋白表达。结果：实时PCR扩增曲线显示结直肠癌组织和正常结直肠组织均有NS基因指数式扩增,癌组织NSmRNA表达量显著高于正常组织（1．8939±0．9529对1．P=0．011）。蛋白质印迹法证实NS蛋白在正常组织中表达较微弱或几乎无表达,癌组织中其表达显著增高（1．5397±0．3625对0．6336±0．3443,P=0．001）。结论：结直肠癌组织和正常结直肠组织中均存在NS基因．推测其微量表达可调节细胞正常增殖．而异常高表达则可通过某些机制促进细胞恶性增殖。     

PTEN、Kindlin-2和DcR3在老年结直肠癌的表达及其对预后的判断价值

目的探讨PTEN、Kindlin-2和DcR3在老年结直肠癌的表达及其对预后的判断价值。方法选取80例健康对照者为对照组,80例结直肠癌患者为观察组,应用流式细胞术检测结肠黏膜组织PTEN、Kindlin-2和DcR3蛋白的表达。比较各组PTEN、Kindlin-2和DcR3表达情况及其与结直肠癌患者不同临床特征的关系,PTEN、Kindlin-2和DcR3蛋白表达量与患者生存的关系。结果相比于对照组,观察组PTEN蛋白表达量明显下降(P0.05);观察组Kindlin-2蛋白表达量明显上升(P0.05);观察组DcR3蛋白表达量明显上升(P0.05)。PTEN、Kindlin-2和DcR3的表达与结直肠癌的淋巴结转移、Dukes分期及增殖细胞核抗原(PCNA)的表达密切相关(P0.05)。结直肠癌患者PTEN(χ2=12.23,P=0.031),Kindlin-2(χ2=7.23,P=0.042)与DcR3(χ2=14.35,P=0.021)与生存密切相关,PTEN蛋白表达阴性、Kindlin-2蛋白表达阳性、DcR3蛋白表达阳性的患者预后较差。结论结直肠癌中PTEN、Kindlin-2和DcR3异常表达对肿瘤的进展有重要意义,联合检测三者表达可能有助于预后判断。     

Cdx1 induced intestinal metaplasia in the transgenic mouse stomach: comparative study with Cdx2 transgenic mice

BACKGROUND AND AIMS: Gastric intestinal metaplasia, which is mainly induced by Helicobacter pylori infection, is thought to be a precancerous lesion of gastric adenocarcinoma. Intestinal metaplastic mucosa expresses intestine specific homeobox genes, Cdx1 and Cdx2, in the human gastric mucosa. We and others have reported that ectopic expression of Cdx2 in the gastric epithelium generates intestinal metaplasia in the transgenic mouse model. METHODS: To clarify the differences in the roles of Cdx1 and Cdx2 in intestinal metaplasia, we generated transgenic mice expressing Cdx1 in the gastric mucosa and compared Cdx1 induced gastric mucosal morphological changes with Cdx2 induced intestinal metaplasia. RESULTS: The gastric mucosa in Cdx1 transgenic mice was completely replaced by intestinal metaplastic mucosa, consisting of all four intestinal epithelial cell types: absorptive enterocytes, goblet, enteroendocrine, and Paneth cells. Paneth cells, which were not recognised in Cdx2 transgenic mice, were in the upper portion of the intestinal metaplastic mucosa. Pseudopyloric gland metaplasia, which was induced in Cdx2 transgenic mice, was not recognised in Cdx1 transgenic mice. Proliferating cell nuclear antigen (PCNA) positive cells were diffusely scattered in Cdx1 induced intestinal metaplastic mucosa while PCNA positive cells in Cdx2 induced intestinal metaplastic mucosa were in the base of the metaplastic mucosa. Intestinal metaplastic mucosa of Cdx1 transgenic mouse stomach was significantly thicker than that of wild-type or Cdx2 transgenic mouse stomach. CONCLUSIONS: We have confirmed that Cdx1 induced gastric intestinal metaplasia but that it differed from Cdx2 induced intestinal metaplasia in differentiation, structure, and proliferation.     

Reprogramming of intestinal differentiation and intercalary regeneration in Cdx2 mutant mice.

The homeobox gene Cdx2, a homologue of the Drosophila gene caudal, has been implicated in the control of cell differentiation in the intestinal epithelium. Recently, we showed that mice in which one allele of the Cdx2 gene had been inactivated by homologous recombination developed multiple intestinal polyp-like lesions that did not express Cdx2 and that contained areas of squamous metaplasia in the form of keratinizing stratified squamous epithelium, similar to that occurring in the mouse esophagus and forestomach. We have now examined colonic lesions from 98 Cdx2+/- mice and report that the lesions are composed of heterotopic stomach and small intestinal mucosa. We conclude that Cdx2 directs endodermal differentiation toward a caudal phenotype and that haploinsufficient levels of expression in the developing distal intestine lead to homeotic transformation to a more rostral endodermal phenotype, such as forestomach epithelium that does not express Cdx2 during normal development. Intercalary growth (epimorphic regeneration), which previously has never been described in mammals, then occurs, resulting in the ordered "filling in" of tissue types at the discontinuity between the gastric and colonic epithelia. This intercalary growth in a restricted space results in the formation of the polypoid lesions observed.     

COX-2、EGFR和Ki67在大肠癌组织中的表达及其意义

目的 探讨环氧合酶-2(COX-2)、表皮生长因子受体(EGFR)和Ki67在大肠癌组织中的表达及其意义.方法 选取大肠癌70例、大肠腺瘤21例和正常大肠组织15例,采用免疫组织化学Envision方法检测COX-2、EGFR和Ki67的表达.结果 在正常大肠黏膜、大肠腺瘤和大肠癌组织中,COX-2的阳性表达率分别为33.3%、71.4%和80.0%,大肠癌和腺瘤组显著高于正常组(P均＜0.05);EGFR的阳性表达率依次为0、23.8%和65.7%,两两比较有统计学差异(P均＜0.05);Ki67的阳性表达率依次为40.0%、76.2%和82.8%,大肠癌和腺瘤组织中的表达均高于正常大肠组织(P均＜0.05).COX-2在有淋巴结转移的大肠癌组中的表达明显高于无淋巴结转移组(P＜0.05);EGFR在低分化、高分期和有淋巴结转移大肠癌中的表达明显增强(P均＜0.05),且COX-2与EG-FR、COX-2与Ki67、EGFR与Ki67之间均具有相关性(r=0.316,0.435,0.314,P均＜0.05).结论 COX-2、EGFR和Ki67对大肠癌的发生、发展有协同作用.     

Cyclooxygenase (COX)-2 immunoreactivity and relationship to p53 and Ki-67 expression in colorectal cancer

The tumor-suppressive effects of nonsteroidal antiinflammatory drugs (NSAIDs) have been suggested to be due to a reduction in cyclooxygenase (COX)-2 activity, although the effects of COX-2 in the colonic mucosa and in colorectal cancer have not been determined. Ki-67 immunoreactivity in cancers is also attracting attention, as Ki-67 reflects cell proliferation, while p53 immunoreactivity is also of interest, as it reflects the malignancy of colorectal lesions. Accordingly, to determine these correlation, we investigated the distribution and intensity of COX-2, p53 and Ki-67 expression in both cancerous and non-cancerous tissues from patients with sporadic and ulcerative colitis (UC)-associated colorectal cancer. We selected 21 colorectal cancer specimens, obtained by surgical resection or colonoscopic biopsy, from 21 patients, including 3 with UC (13 men and 8 women; aged 42–78 years). Histological examination of hematoxylin and eosin-stained specimens revealed that 9 were well differentiated; 11, moderately differentiated; and 1 was a poorly differentiated adenocarcinoma. We used anti-COX-2, p53, and Ki-67 antisera to perform immunohistochemical staining by the labelled streptavidin biotin method and then assessed and graded the staining intensity and distribution. COX-2 staining was more intense in cancer tissue than in non-cancerous areas. Colorectal cancers associated with UC were not stained intensely. COX-2, p53, and Ki-67 positivity rates in were 38.1%, 38.1%, and 47.6%, respectively. There were no relationships among the distributions or intensities of COX-2, p53, and Ki-67 expression. Our results indicate that colorectal cancer tissues overexpress COX-2, but that there are no relationships between COX-2, p53, and Ki-67 expression, suggesting that COX-2 expression may not be related to cell proliferation or to the grade of malignancy. However, it is necessary to determine whether COX-2 in cancer tissue is involved in carcinogenesis or whether it is simply a product of cancer. (Received: July 7, 1998; accepted: Oct. 23, 1998)     

环氧化酶2在大肠癌中的表达及其与大肠癌生物学特性的关系

[目的]通过观察环氧化酶2（Cyclooxygenase-2,COX-2）在大肠腺瘤、大肠癌中的表达及其与大肠癌生物学特性的关系,初步探讨其在大肠癌发生、发展过程中的作用机制。[方法]应用免疫组织化学染色法对78例大肠癌组织、21例大肠腺瘤组织和13例正常大肠黏膜组织进行免疫组化染色;应用它检验分析COX-2的表达情况及其与大肠癌临床病理特征的关系。[结果]大肠癌组织中COX-2阳性表达率为78．21％,明显高于大肠腺瘤的52．38％和正常大肠黏膜组织的7．69％（P〈0．05）;COX-2表达与大肠癌的Duke＇s分期、分化程度、淋巴结转移有相关性（P〈0．05）,而与大肠癌患者的性别、组织学类型无关（P〉0．05）。[结论]COX-2在大肠癌组织中表达率明显高于大肠腺瘤,而正常大肠黏膜中表达率极低或不表达;COX-2表达与大肠癌生物学特性有明显相关性。