大肠腺瘤与大肠腺癌肿瘤相关基因的差异表达

目的:探索大肠腺瘤与大肠腺癌差异表达的肿瘤相关基因.方法:利用基因芯片技术筛选大肠腺瘤与大肠腺癌差异表达的肿瘤相关基因,比较两组基因特点,寻找大肠腺癌重要致病基因,并以RT-PCR对部分差异表达基因进行检测来验证芯片结果.结果:(1)大肠腺瘤差异表达的肿瘤相关基因9个,均上调;(2)大肠腺癌差异表达的肿瘤相关基因47个...     

大肠腺瘤细胞癌变过程中差异表达蛋白的研究进展

大肠癌是消化道常见的恶性肿瘤,而大肠腺瘤被认为是大肠癌的癌前病变之一,腺瘤-腺癌序列(正常大肠上皮-腺瘤-腺癌)也被认为是经典的大肠癌发病过程.研究表明,大肠腺瘤恶变的过程中,多种蛋白的表达水平会发生变化,某些蛋白发生结构的变化,这些蛋白涉及许多种如细胞内酶蛋白、骨架相关蛋白、信号蛋白等.其致癌机制也有很大差别,并且还未完全明确.本文就近年来这些差异表达的蛋白质进行综述,并对其可能的致癌机制进行简要描述.近年来蛋白组学技术也得到了快速发展,由于其检测手段先进,高通量而且可以同时检测多种蛋白,已经被逐步应用于差异蛋白的检测和定性,所以本文就大肠腺瘤与腺癌之间的差异蛋白组学也做了简要介绍,有助于早期发现大肠癌的标志物,早期诊断及了解其发病机制.     

CD44 expression in benign and malignant colorectal polyps

PURPOSE: This retrospective study was undertaken to evaluate immunohistochemically the expression of CD44 standard protein and CD44v5 and CD44v6 isoforms in colorectal adenomas and early invasive cancers developing within adenomas as possible markers characterizing colorectal polyps with a more aggressive biologic potential. METHODS: Archival tissues of 81 consecutive locally resected colorectal polyps, comprising 57 colorectal adenomas and 24 carcinomas-in-adenomas, were stained immunohistochemically with the use of commercially available mouse monoclonal antibodies: SFF-2 for CD44 standard protein, VFF-8 for CD44v5, and VFF-7 for CD44v6. RESULTS: Sixtythree percent of the colorectal polyps were positive for CD44 standard protein, 59 percent were positive for CD44v5, and 27 percent were positive for CD44v6. Ninetythree percent of the low-grade adenomas were CD44 standard protein-positive, in contrast to 50 percent of the high-grade adenomas and only 42 percent of the carcinomas-in-adenomas (Kendall's Tau =–0.42;P<0.0001). CD44v6 expression was more frequently found in early invasive cancers (54 percent) than in high-grade adenomas (25 percent) and low-grade adenomas (7 percent). This difference also was statistically significant (Kendall's Tau-b =0.39;P=0.00003). Surprisingly, a downregulation of CD44 standard protein expression was observed in the adenoma tissue adjacent to carcinomas (62 percent) and areas with high-grade atypia (71 percent), compared with low-grade adenomas (93 percent; Kendall's Tau-b =–0.28;P=0.004). CONCLUSIONS: Our data suggest that CD44 standard protein and CD44 isoform v6 expression differs considerably in benign and malignant colorectal polyps. Clinical studies with larger patient groups could clarify the prognostic potential of CD44 further     

Bcl-2与PCNA在大肠腺瘤和大肠癌中的表达

目的　探讨增殖细胞核抗原 (proliferatingcellnuclearantigen ,PCNA)与原癌基因bcl 2在大肠腺瘤和大肠癌中的表达变化以及与大肠腺瘤和大肠癌的发生、发展的关系。方法　采用免疫组化S P法 ,分别检测PCNA与bcl 2在 18例大肠腺瘤和 18例大肠癌中的表达。结果　PCNA在大肠腺瘤和大肠癌中的标记指数分别为 65 95± 12 94%和 84 5 3± 10 3 9% ,两者均明显高于正常肠粘膜中的指数 ,P <0 0 0 1,两者相比有显著性差异 ,P <0 0 1。Bcl 2在大肠腺瘤和大肠癌中的表达分别为 81 10± 10 0 0 %和 65 78±12 95 % ,两者均明显高于正常肠粘膜中的表达P <0 0 0 1。两者相比有显著性差异 ,P <0 0 1。结论　大肠癌的癌前病变腺瘤中存在活跃的细胞增殖 ,而且同时有细胞凋亡的改变 ,细胞增殖和细胞凋亡的调控失常共同导致了肿瘤的发生。     

Differential expression of the carcinoembryonic antigen-related cell adhesion molecules panCD66, CD66a,CD66c and of sialyl-Lewis x (CD15s) on blast cells of acute leukemias

Expression of CD66 has been reported to occur on blast cells from children with acute lymphoblastic leukemia (ALL), but little is known about the differential expression pattern of panCD66 and other members of the CD66 family on blast cells from patients with acute myeloid leukemia (AML). We have performed flow cytometry immunophenotyping on blast cells from 28 patients with acute myeloid leukemia (AML), 13 patients with B-cell precursor acute lymphoblastic leukemia (BCP-ALL) and 7 patients with T-ALL using monoclonal antibodies (mAbs) against panCD66 (clone D14HD11), CD66a (clone 4.3.17), CD66c (clone 9A6) and CD15s. Expression of the panCD66 mAb was found to be positive in 13 of 28 patients with AML (46%) and in 6 of 13 patients with BCP-ALL (46%) but negative for all the seven patients with T-ALL. In AML, panCD66, CD66a and CD66c were more frequently coexpressed with CD65, CD15 and CD64 than with CD13, CD33 or the two progenitor markers CD34 and CD117. In contrast to CD15, the expression of the sialylated Lewis X (CD15s) was associated with CD117 positivity in the majority of AML cases (64 vs. 85%, P = 0.043). Radioimmunotherapeutic strategies targeting CD66 antigens should consider the heterogeneous expression pattern of CD66 molecules in acute leukemias especially in AML where expression is correlated with mature granulomonocytic cells but not with CD34 and CD117 positive progenitor cells.     

CD66 expression in acute leukaemia

Antibodies against CD66 identify antigens from the carcinoembryonic antigen (CEA) family of proteins, which belong to the immunoglobulin gene superfamily. Despite being usually restricted to cells of myeloid or monocytic origin, CD66 expression has also been reported in blasts from children with B-cell lineage acute lymphocytic leukaemia (ALL). An analysis of the CD66 expression was undertaken in a series of acute leukaemia patients. Antigenic expression was analysed using triple combinations of monoclonal antibodies (mAbs) in forty-five patients. The CD66 Kat4 fluorescein isothiocyanate clone was purchased from Dako (Glostrup, Denmark). CD66 was expressed in 2 of 29 patients with AML (acute myeloblastic leukemia) (6.8%) and in 8 of 12 patients with B-cell lineage ALL (66.7%; P <0.001); in blast crisis (BC) of chronic myelocytic leukaemia (CML), CD66 was expressed in two patients with lymphoid BC but not in the two with myeloid BC. The co-expression of CD66 with other myeloid antigens was observed in all CD66+ ALL/Ly-BC cases tested: CD 13 in six patients, CD33 in seven and CD117 in two patients. The CD66 expression is more frequent in ALL than in AML. Furthermore, we analysed minimal residual disease (MRD) in eight patients in complete remission. CD66 expression was associated with an abnormal B-cell differentiation pattern and with increases in CD34/CD19+ cells in all but one case. These findings suggest that an aberrant expression of CD66 could be used to investigate MRD in ALL. The association between CD66 reactivity and bcr-abl in adult ALL remains to be investigated. Received: 31 May 1999 / Accepted: 10 November 1999     

结肠腺瘤和结肠癌基因表达谱分析

目的 研究结肠腺瘤、结肠癌基因表达谱,筛查结肠癌相关基因.方法 用BRB-Array Tools对公共基因芯片数据库GEO中的结肠腺瘤、结肠癌基因芯片表达数据进行统计学分析,找出在结肠腺瘤和结肠癌发生发展中均发生变化的基因及二者的特异性基因,进一步分析其功能.结果 样本聚类表明各类组织分类正确,比较后得到在腺瘤和腺癌中共同表达的104条基因,其中48条共同上调,56条共同下调;差异基因的功能涉及物质代谢、内环境稳态、细胞间通讯、细胞黏附、细胞增殖等多种肿瘤发生发展的重要生物学过程.结论 生物信息学方法发现的结肠腺瘤、结肠癌基因表达谱可为结肠癌的发病机制及治疗靶位的研究开辟新思路.     

Human chorionic gonadotrophin expression in colorectal adenocarcinoma

The presence of human chorionic gonadotrophin (HCG) in colorectal adenocarcinoma was studied histologically in 45 tumors using immunoperoxidase technique. Ten neoplasms (22.2 percent) contained HCG-positive tumor cells. These cells were present mostly at the periphery of the tumors. Many formed parts of glands, while some were arranged in syncytial clumps or columns, or singularly. Thus, these tumor cells resembled trophoblastic tissue not only in being HCG-positive but also in their peripheral distribution and occasionally in morphologic appearance. HCG-positive tumors were seen more commonly in the rectosigmoid region (90 percent) and were more aggressive than HCG-negative tumors. In this study, lymph node or liver metastases were present in 70 percent of HCG-positive tumors compared with 29 percent of negative tumors—a difference which is statistically significant.     

大肠锯齿状腺瘤基础与临床研究进展

由于中国人生活方式和饮食习惯的改变等原因，大肠癌的发病率也呈逐年上升趋势。目前普遍认同大肠腺瘤是大肠上皮从良性向恶性演变的重要环节，也是最具恶变潜能的癌前状态。国内外学者对大肠腺瘤进行着广泛而深入地研究。近年来大肠锯齿状腺瘤（serrated adenoma，简称锯齿状腺瘤）正日益受到关注。大肠锯齿状腺瘤独特的结构特征和细胞学特点由Goldman等于1970年最先报道，1990年Longacre等报道了110例，并首次将这类病例命名为大肠锯齿状腺瘤。据报道大约5％～10％的锯齿状腺瘤病灶内发生黏膜内癌。2000年世界卫生组织（WHO）正式将大肠锯齿状腺瘤定为大肠的第四种腺瘤病理形态（前三种是管状腺瘤、     

Flow cytometric identification of myeloid disorders by asynchronous expression of the CD14 and CD66 antigens

Abstract: Using a multiparameter flow cytometry assay enumerating cells positive for CD13, CD14 and CD66 antigens, we determined the asynchronous CD14/CD66 co-expression in unselected bone marrow and peripheral blood samples with suspected malignant blood disorders. CD14/CD66 co-expression >5% were found in 131/691 bone marrow samples. Only 55 of these exhibited an identifiable population in 2-parameter flow cytometry histograms. Of the 55 samples 43 (78%) came from patients with myeloid disorders; e.g. 11 with myelodysplastic syndromes, 15 with chronic myeloproliferative disorders and 17 with acute myeloid leukaemia. Only one of these 17 cases was a de novo case, while 8 were secondary to another malignant haematological disease and 8 were from the period after cytoreductive therapy. Notably, CD14/CD66 co-expression patterns were related to disease categories; e.g. in chronic myelomonocytic leukaemia and acute myeloid leukaemia following a dysplastic phase the co-expression displayed two subsets in peripheral blood, low-avidity CD14 and low-avidity CD66, respectively. The latter disease category also exhibited these 2 subsets in bone marrow. In all other cases, the CD14/CD66 co-expression in bone marrow was heterogeneous. In conclusion, abnormal CD14/CD66 co-expression might be a valuable parameter in defining asynchronous myelopoiesis in malignant myeloid disorders, especially myeloproliferative disorders and secondary acute myeloid leukemias.     

CD83、CD1a和Ki-67在大肠癌中的表达及意义

目的:探讨CD83、CD1a和Ki-67在大肠癌组织中的表达及预后意义.方法:采用免疫组织化学和流式细胞术检测60例大肠癌组织中CD83、CD1a和Ki-67的表达情况,并分析其与大肠癌临床病理特点和预后的关系.结果:免疫组织化学检测显示,低Dukes分期、预后好的癌组织中CD83和CD1a阳性率均显著高于对应组(均P<0.05),而FCM检测只显示CD1a阳性率显著高于对应组(均P<0.05);两检测均显示,高Dukes分期、预后差的癌组织中Ki-67阳性率显著高于对应组(均P<0.05).结论:CD83、CD1a和Ki-67的表达与大肠癌Dukes分期和预后相关,可以作为检测大肠癌预后的参考指标.     

Cancer stem cell markers CD133 and CD24 correlate with invasiveness and differentiation in colorectal adenocarcinoma

AIM： To verify that CD markers are available for detecting cancer stem cell populations and to evaluate their clinical significance in colon cancer.METHODS： Immunohistochemistry for CD133, CD24 and CD44 was performed on the tissue microarray of 523 colorectal adenocarcinomas. Medical records were reviewed and clinicopathological analysis was performed.RESULTS： In colorectal adenocarcinoma, 128 of 523 cases （24.5%） were positive and 395 cases （75.5%） were negative for CD133 expression. Two hundred and sixty-four of 523 cases （50.5%） were positive and 259 cases （49.5%） were negative for CD24 expression. Five hundred and two of 523 cases （96%） were negative and 21 cases （4%） were positive for CD44 expression. Upon clinicopathological analysis, CD133 expression was present more in male patients （P = 0.002） and in advanced T stage cancer （P = 0.024）. Correlation between CD24 expression and clinicopathological factors was seen in the degree of differentiation （P= 0.006）. Correlation between CD44 expression and clinicopathological factors was seen in the tumor size （P = 0.001）. Survival was not significantly related to CD133, CD24 and CD44 expression.CONCLUSION： CD markers were related to invasiveness and differentiation of colorectal adenocarcinoma. However, CD expression was not closely related to survival.     

结直肠腺瘤患者血脂及脂蛋白变化的研究

目的　探讨血脂及脂蛋白变化与结直肠腺瘤的关系。方法　结直肠腺瘤患者 180例 ,对照组 5 5例 ,收集清晨静脉血 2ml,分别测定血清总胆固醇 (TC)、甘油三脂 (TG)、高密度脂蛋白胆固醇 (HDL C)和低密度脂蛋白胆固醇 (LDL C) ,并根据腺瘤的部位、大小、数量、组织成分、异型增生的情况进行分组 ,分别比较各组的测定值。结果　腺瘤组TC、TG、LDL C较对照组显著升高 (P <0 .0 1) ,HDL C显著下降 (P <0 .0 1) ,其他各亚组测定值也有相应改变。结论　TC、TG、LDL C与腺瘤的发生及恶变有关 ,HDL C对其有抑制作用。     

Current and future molecular diagnostics in colorectal cancer and colorectal adenoma

Colorectal cancer(CRC)is one of the most prevalent cancers in developed countries.On the other hand,CRC is also one of the most curable cancers if it is detected in early stages through regular colonoscopy or sigmoidoscopy.Since CRC develops slowly from precancerous lesions,early detection can reduce both the incidence and mortality of the disease.Fecal occult blood test is a widely used non-invasive screening tool for CRC.Although fecal occult blood test is simple and cost-effective in screening CRC,there is room for improvement in terms of the accuracy of the test.Genetic dysregulations have been found to play an important role in CRC development.With better understanding of the molecular basis of CRC,there is a growing expectation on the development of diagnostic tests based on more sensitive and specific molecular markers and those tests may provide a breakthrough to the limitations of current screening tests for CRC.In this review,the molecular basis of CRC development,the characteristics and applications of different non-invasive molecular biomarkers,as well as the technologies available for the detection were discussed.This review intended to provide a summary on the current and future molecular diagnostics in CRC and its pre-malignant state,colorectal adenoma.     

Measurement of circulating biliary glycoprotein (CD66a) in liver diseases

Purpose. Biliary glycoprotein (BGP), a member of the carcinoembryonic antigen (CEA) gene family, is produced by hepatocytes, and is suggested to function as a cell adhesion molecule, mouse hepatitis virus receptor, and tumor suppressor. Our aim was to establish an enzyme immunoassay for circulating BGP and to study its significance in liver diseases. Methods. For enzyme immunoassay, a monoclonal antibody (mAb), TS135, which recognizes BGP was used as a catcher, and biotin-labeled polyclonal anti-CEA antibodies were used as a tracer. Seventy-six serum specimens obtained from patients with various liver diseases were submitted to the assay. Results. The incidence of positivity for antigen TS135 in the serum samples of the 76 patients was 57.9%. The most significant correlation among conventional liver function tests was found between antigen TS135 and γ-glutamyl transpeptidase (γ-GTP). However, among the 56 patients whose serum antigen TS135 and γ-GTP levels could be measured simultaneously, 5 were antigen TS135-positive and γ-GTP-negative (8.9%) and 6 were antigen TS135-negative and γ-GTP-positive (10.7%). The increased serum level of antigen TS135 in 6 cholangiocellular carcinoma (CCC) patients led us to the immunohistochemical study of CCC, in which 8 of the 8 tissue specimens tested were positive for mAb TS135, indicating the production of the antigen from CCCs. Conclusions. This preliminary study suggests that the circulating antigen TS135 level correlates with γ-GTP in liver diseases, but that TS135 may also have a unique significance, different from that of γ-GTP, as a liver function test. Received: September 14, 2000 / Accepted: January 26, 2001     

410例患者结直肠腺瘤与幽门螺杆菌感染特征及相关性分析

目的　探讨结直肠腺瘤与幽门螺杆菌感染的关系。 方法　连续性收集在上海交通大学医学院附属仁济医院内镜中心同时行胃镜及结加肠镜的患者共410例患者的病例资料（其中结直肠腺瘤者74人,正常对照组336人）。比较结直肠腺瘤组和健康对照组幽门螺杆菌感染情况的差异。并将结直肠腺瘤组分别按腺瘤的数目、最大径、病理分型分亚组,比较各亚组与幽门螺杆菌感染情况。统计学分析采用t检验与卡方检验,P < 0.05 被认为具有统计学差异。 结果　结直肠腺瘤组和健康对照组幽门螺杆菌感染阳性率分别为29.73%和19.34%,前者高于后者,差异均有统计学意义（P＜0.05）。按腺瘤数目、最大径、病理分型分亚组,各亚组组内幽门螺杆菌感染阳性率差异均无统计学意义（P均＞0.05）,但按递进分层与对照组相比均有统计学差异（P均小于0.05）。 结论　结直肠腺瘤可能与幽门螺杆菌感染有直接相关,同时幽门螺杆菌对腺瘤的发展可能有促进作用。     

CADM1表达下调与大肠腺癌发生发展的关系

目的探讨CADM1表达下调与大肠腺癌发生发展的关系。方法RT—PCR检测正常大肠组织、大肠腺瘤和大肠腺癌的CADM1mRNA表达;免疫组化法检测正常大肠组织、大肠腺瘤和大肠腺癌的CADMI蛋白表达。结果大肠腺瘤中CADM1表达无缺失或明显减少（P〉0．05）;部分大肠腺癌组织中CADM1表达下调,且有淋巴结转移患者比无淋巴结转移者更常见（P〈0．05）,Dukes’C期和Dukes’D期患者比Dukes’A期和Dukes’B期患者更常见（P〈0．05）。结论CADM1表达下调在大肠腺癌中常见,特别是在进展期大肠腺癌,因此,CADM1在大肠腺癌发病机制中可能发挥重要作用。     

The analysis of mononuclear cell infiltrations in colorectal adenocarcinoma

Summary Mononuclear cell (MNC) populations in tissue specimens from 11 colorectal adenocarcinomas and 1 mucinous adenocarcinoma were analyzed, applying different monoclonal antibodies (moAB). Tumor epithelium could be characterized by MoAB VEP9, predominantly binding to mucus secreted by the tumor cells. In approximately 30%–50%, the tumor epithelium also reactdd in a patchy pattern with the MoABs OKT10 and OKIa, which define in the peripheral blood activated and HLA-DR expressing cells. T-lymphocytes, as defined by the MoAB 9.6, and T-lymphocyte subpopulations, as characterized by the MoABs OKT4 and OKT8, were found in the intratumoral stroma and in peritumoral inflammatory areas. Quantifying the relative amounts of mononuclear cell subpopulations in the different stroma compartments, a predominance of OKT10- and OKIa-defined and MoAB-S39-reactive cells was observed. NK cells, as labelled by the MoAB Leu-7, were only demonstrated singularly in different areas of the stroma. Nonlymphoid structures such as vessel walls were shown to express antigens recognized by the MoABs OKIa, OKT10, and Leu-7. In some instances, nerve structures were labelled by the MoABs OKIa, OKT10, S39, and Leu-7. Using semiquantitative analysis, no correlation could be obtained between the intratumoral and peritumoral infiltration with T-lymphocytes, T-lymphocyte subpopulations and monocytic cells, and histopathological tumor grading and staging.Abbreviations MoAB monoclonal antibody - MNC mononuclear cell(s) - PBS phosphate buffered saline - PGE2 prostaglandin E 2 Supported by DFG, SFB 118     

MPP3表达减少与大肠癌发病的关系

目的研究MPP3表达减少与大肠癌发病的关系。方法 RT-PCR法检测正常大肠黏膜、大肠腺瘤和大肠癌的MPP3 mRNA表达;Western Blotting法检测正常大肠黏膜、大肠腺瘤和大肠癌的MPP3蛋白表达。结果大肠腺瘤中MPP3表达无明显减少(P>0.05),部分大肠癌组织中MPP3表达减少,且有淋巴结转移患者比无淋巴结转移患者更常见(P<0.05),Dukes C期和D期患者比Dukes A期和B期患者更常见(P<0.05)。结论 MPP3表达减少在大肠癌中为常见现象,而且在晚期肿瘤更明显,因此,MPP3在大肠癌发病中可能发挥重要作用。     

DNA甲基转移酶1和3A在结直肠癌组织中的表达及其与临床病理的关系

目的:探讨DNA甲基转移酶1(DNMT1)和DNA甲基转移酶3A(DNMT3A)在结直肠正常组织、腺瘤、腺癌中的表达,及其可能的临床病理意义.方法:以免疫组织化学SP法检测DNMT1和DNMT3A在正常结直肠组织、腺瘤、腺癌中的表达.通过计算染色指数(SI),评估两者的表达水平与年龄、肿瘤大小、分期、分化等临床病理特征之间的关系.采用Kaplan-Meier法进行生存分析.结果:DNMT3A在正常组织和腺瘤中表达明显低于癌组织(SI:10.5±5.7vs20.2±9.9,P<0.05).DNMT1在三者中的表达呈递增趋势,SI分别为10.3±2.7、14.7±6.8、20.1±9.1.淋巴结转移病例DNMT1表达高于未转移病例(18.1±7.9vs12.0±6.3,P<0.05),浸润深度超过肌层病例DNMT1和DNMT3A表达高于未超过肌层病例,差异均有统计学意义(20.1±10.1vs15.8±7.9;19.9±8.5vs15.4±4.7,均P<0.05).结论:DNMT1和DNMT3A是结直肠癌发生的早期事件,两者共同促进结直肠癌的发生发展,可能成为结直肠癌治疗的新靶点.