大肠癌中survivin基因的表达及相关性研究

目的　探讨survivin凋亡抑制基因的表达变化与大肠癌生物学行为及cyclinD1、bcl 2、p5 3和凋亡指数 (AI)的相关性。方法　利用免疫组织化学EnVision法检测 98例大肠癌、2 4例正常大肠黏膜和 2 9例大肠息肉组织中survivin、bcl 2、cy clinD1,并用TUNEL法测定AI。结果　survivin、bcl 2、cyclinD1和 p5 3在 98例大肠癌组织中的阳性表达率分别为 6 8 37%、6 0 2 %、4 0 8%和 5 2 %。survivin表达在癌细胞胞质内 ,核内未见表达 ,bcl 2表达在细胞质内 ,cyclinD1和 p5 3均在核内表达。survivin的表达与组织学类型和淋巴结转移密切相关 (P均 <0 0 5 ) ,其阳性表达组的AI显著低于阴性组 (P <0 0 1) ,其阳性表达者术后 5年复发转移率均较阴性者显著增高 (P <0 0 1) ,而 5年生存率则显著降低 (P <0 0 5 ) ,survivin表达与bcl 2、p5 3无相关性 ,与cyclinD1表达呈正相关 (P <0 0 5 )。bcl 2和cyclinD1的阳性表达与组织学类型关系密切 (P均 <0 0 5 ) ,bcl 2阳性组的AI明显低于阴性组 (P <0 0 5 ) ,cyclinD1的阳性表达与 5年生存率明显相关 (P <0 0 5 )。 结论　survivin、bcl 2、cy clinD1和p5 3基因在大肠癌的发生、发展中起着不同程度的作用 ,它们的检测对大肠癌恶性程度的判定 ,预后分析和进一步治疗提     

结直肠癌nm23基因表达、突变与侵袭、转移的相关性

目的　探讨nm2 3基因异常表达及突变与结直肠癌侵袭、转移之间的关系。方法　应用免疫组化LDP法、流式细胞术、PCR SSCP法对 1 5 0例结直肠癌标本及正常黏膜组织和 1 6例大肠腺瘤分 3组进行nm2 3蛋白检测和基因突变的筛查。结果　①免疫组化发现 73例结直肠癌中nm2 3 H1蛋白表达水平低于正常黏膜组织和良性腺瘤 (P <0 0 0 1 ,P <0 0 0 1 ) ,转移癌表达低于原发癌 (P <0 0 0 1 ) ,结直肠癌nm2 3 H1蛋白表达与组织分化、肠壁浸润、Dukes分期、转移均密切相关 (P <0 0 0 1 ) ,低表达组术后预后差于高表达组 (P <0 0 0 1 ) ;②流式细胞术研究表明 ,nm2 3 H1蛋白在癌组织表达低于癌旁组织 ;在结直肠癌伴有转移 (3/ 1 4 )时其表达低于未发生转移者 (1 1 / 1 4 ) ;③nm2 3 H1基因第 2～ 5位外显子在 6 3例结直肠癌组织中未发现异常突变。结论　结直肠癌中nm2 3基因突变可能是小概率事件 ,在结直肠癌的侵袭、转移中不发挥主要作用 ,而nm2 3 H1蛋白表达降低则与结直肠癌的侵袭、转移有关 ,提示检测nm2 3蛋白表达是判断结直肠癌预后有价值的指标之一。     

结直肠癌中cyclin D1和p27蛋白的表达及其意义

目的　研究cyclinD1和p2 7蛋白在结直肠癌发生、发展中的作用及其与结直肠癌临床病理特征关系。 方法　收集5 8例手术切除的结直肠癌标本 ,同时取距癌灶 >5cm的正常组织 ,应用免疫组化S P法检测癌组织及正常组织中cyclinD1和p2 7蛋白的表达。结果　cyclinD1蛋白在结直肠癌的表达阳性率为 5 5 17%,正常组织无表达 (P <0 0 1) ;cyclinD1蛋白的表达阳性率在 6 0岁以上年龄组高于 6 0岁以下年龄组 (P <0 0 5 ) ;cyclinD1蛋白的表达与肿瘤组织分化程度负相关 (P <0 0 1)。p2 7蛋白在结直肠癌的表达阳性率为 5 5 17%,在结直肠正常组织的表达阳性率为 96 5 5 %(P <0 0 1) ;p2 7蛋白的表达与肿瘤组织分化程度负相关 (P <0 0 1)。cyclinD1和p2 7蛋白在结直肠癌的表达呈正相关 (r =0 5 82P <0 0 1)。 结论　cyclinD1蛋白过表达与 p2 7蛋白失活可加速细胞周期转化 ,促进结直肠癌的发生 ,cyclinD1和 p2 7蛋白的检测可作为评价结直肠癌恶性程度和判断预后的重要指标。     

肿瘤增殖和凋亡相关因素在胃癌演进过程中的作用

目的　探讨肿瘤增殖和凋亡相关因素在胃癌发生、发展中的相互作用。方法　采用免疫组化双重染色法对 15 6例正常胃黏膜、不典型增生及胃癌标本分别进行EGFR、Ki 6 7、bcl 2和p5 3的染色。分析两个增殖相关因素 (EGFR、Ki 6 7)之间和凋亡相关因素 (bcl 2、p5 3)之间的相关性。 结果　 (1)上述 4者在正常胃黏膜中均呈阴性表达 ,EGFR在重度不典型增生时呈第一高峰 ,Ⅰ级癌时表达下降 ,Ⅱ～Ⅲ级癌时呈现第二高峰 ,Ki 6 7在胃癌中的表达率 (76 6 % )高于不典型增生 (4 1 4 % ) (P <0 0 1) ,bcl 2在重度不典型增生和Ⅰ级癌中的表达高于轻、中度不典型增生和Ⅱ～Ⅲ级癌 (P <0 0 5 ) ,p5 3在胃癌中的表达高于不典型增生 ,且表达率随分化程度的降低而升高 (P <0 0 5 )。 (2 )EGFR、Ki 6 7和 p5 3的表达与胃不典型增生程度、胃癌分化程度、组织学类型有关 ,而且 ,EGFR和P5 3与浆膜浸润和淋巴结转移有关 ,Ki 6 7虽未达统计学标准 ,但也有此倾向。 (3)EGFR和Ki 6 7表达呈正相关 ,而bcl 2和 p5 3表达呈负相关 (P <0 0 1)。结论　EGFR和Ki 6 7在胃癌发生、发展中有协同作用 ,可作为反映胃癌生物学行为的指标之一。bcl 2和p5 3分别作用于胃癌发生、发展的不同阶段 ,有助于胃癌的早期诊断和预后判断。     

结直肠癌中TGF-β1表达与肿瘤浸润转移和血管形成的关系

目的　探讨结直肠癌中转化生长因子 β1(TGF β1)的表达与肿瘤浸润转移和血管形成的关系。 方法　免疫组化S P法检测 12 6例结直肠癌中TGF β1和血管内皮生长因子 (VEGF)的表达 ,同时应用CD34标记肿瘤间质中微血管密度 (MVD)。结果　 12 6例结直肠癌中TGF β1和VEGF的阳性表达率分别为 4 2 1%和 6 3 5 %。结直肠癌中TGF β1、VEGF蛋白的表达和MVD与肿瘤的浸润深度、淋巴结转移和Dukes分期呈正相关 (P <0 0 5 ) ;VEGF在TGF β1表达阳性的结直肠癌中的阳性表达率高于TGF β1表达阴性的结直肠癌 (P <0 0 5 ) ,TGF β1表达阳性的结直肠癌MVD高于TGF β1表达阴性的结直肠癌(P <0 0 5 )。结论　TGF β1可能通过间接或直接刺激肿瘤血管形成而促进结直肠癌的浸润转移     

甲状腺非典型腺瘤生物学特性及免疫组化研究

目的 :探讨甲状腺非典型腺瘤免疫组化表型与生物学特性的关系。方法 :采用免疫组化S P法 ,检测bcl 2、p5 3、PCNA的表达 ,并与单纯腺瘤和滤泡状癌对照研究。结果 :非典型腺瘤bcl 2、p5 3、PCNA阳性表达率分别为 82 8% (2 3/ 2 8)、2 5 % (7/2 8)、76 5 % (2 1/ 2 8) ,与腺瘤差异有显著性 (P <0 0 1) ,与滤泡状腺癌差异无显著性 (P >0 0 5 )。结论 :甲状腺非典型腺瘤似原位癌 ,手术治疗效果良好 ,罕见复发 ,无转移     

CD44v6和E-cadherin表达与结直肠癌浸润转移关系

目的　探讨CD44v6和E cadherin(E cad)蛋白表达与结直肠癌浸润转移的相关性。方法　应用免疫组织化学技术 ,检测 90例结直肠癌组织中CD44v6和E cad蛋白表达。结果　 90例结直肠癌中CD44v6和E cad蛋白阳性表达率分别为75 6 %和 46 7%。CD44v6高表达及E cad低表达与结直肠癌Dukes分期、浆膜浸润、淋巴结转移、肝脏转移均呈正相关 (P <0 0 5 )。结直肠癌中CD44v6表达与E cad表达呈负相关 (r =- 0 4 3,P <0 0 0 5 )。结论　CD44v6和E cad表达与结直肠癌浸润转移密切相关。检测CD44v6和E cad蛋白表达可作为判断结直肠癌预后的客观指标。     

结直肠癌中SIRT1和Survivin的表达及临床意义

目的观察沉默信息调节因子1(silent information reg-ulator 1,Sir1))、凋亡抑制基因Survivin在结直肠癌中的表达,分析SIRT1、Survivin与结直肠癌(colorectal cancer,CRC)患者临床病理学特征的关系以及二者表达间的关系。方法应用免疫组化SP法检测60例结直肠癌组织和24例正常结直肠黏膜中SIRT1、Survivin的表达。结果 SIRT1、Sur-vivin在结直肠癌中的阳性率均高于正常结直肠黏膜(χ2=15.674,P=0.000;χ2=32.747,P=0.000)。SIRT1表达与结直肠癌浸润深度(P=0.021)、淋巴结转移(P=0.020)、pTNM分期(P=0.003)、p53蛋白表达(P=0.024)均呈正相关。Survivin表达与pTNM分期呈正相关(P=0.035)。SIRT1与Survivin表达无明显相关性(P>0.05)。结论SIRT1、Survivin在结直肠癌组织中过表达,且与多项临床病理指标有关,二者可能成为判定CRC恶性程度及评估预后的生物学指标。     

结直肠癌Stat3表达与细胞增殖、凋亡及预后的关系

目的探讨结直肠癌Stat3蛋白表达与细胞增殖、凋亡、临床病理特征及预后的关系。方法应用免疫组织化学及DNA末端标记(TUNEL)技术分别检测129例结直肠癌及癌旁组织芯片中Stat3、Cyclin D1蛋白表达及凋亡指数(AI),分析Stat3表达与临床病理特征和预后的关系及其与增殖、凋亡之间的相关性。结果结直肠癌组织中Stat3、Cyclin D1阳性表达率分别为67.4%和79.8%,均高于癌旁组织的43.4%和5.4%(P0.05),结直肠癌组织AI(5.62)低于癌旁(7.86)(P0.05)。Stat3的表达与结直肠癌的Dukes分期、淋巴结转移、肿瘤发生部位明显相关(P0.05)。结直肠癌组织中Stat3和Cyclin D1的表达呈正相关(P0.05)。Kaplan-Meier生存分析显示Stat3、Dukes分期、淋巴结转移及凋亡指数与生存期相关(P0.05);Cox回归分析显示Dukes分期为独立的风险预后因子。结论 Stat3可能参与了结直肠癌的发生发展,检测Stat3可作为评估结直肠癌患者预后及淋巴结转移的指标。     

卵巢肿瘤组织中端粒酶逆转录酶的原位表达

目的　探讨卵巢肿瘤组织中端粒酶逆转录酶 (TRT)的表达及其与凋亡基因 p5 3蛋白和bcl 2基因的关系。方法　应用免疫组织化学S P法检测 78例卵巢肿瘤组织中TRT及p5 3、bcl 2的表达 ,结果进行统计学分析。结果　TRT在不同卵巢肿瘤组中的表达差异有显著性 ,在卵巢上皮组织肿瘤中良性、交界性和恶性肿瘤组的阳性表达分别为 6 7%、5 7 2 %和 85 % ,在卵巢生殖细胞肿瘤中良性与恶性的阳性表达为 10 %和 81 8% ,TRT的表达随着肿瘤恶性度增高呈现阳性率和阳性强度递增现象 ,两者比较差异有显著性 (P <0 0 1) ;卵巢恶性上皮组织肿瘤中 p5 3蛋白和bcl 2基因的阳性表达率分别为 90 %和80 % ,相关分析显示TRT与 p5 3和bcl 2的表达密切相关 (P <0 0 1)。结论　TRT的高表达与卵巢恶性肿瘤有密切关系 ,卵巢肿瘤组织中TRT与p5 3、bcl 2的表达呈正相关 ,TRT可作为卵巢肿瘤恶性度的一个指标。     

Coexpression of MUC1 with p53 or MUC2 correlates with lymph node metastasis in colorectal carcinomas

The alteration of the mucin profile have been known to play a role in colorectal carcinogenesis. MUC1 is up-regulated and MUC2 is down-regulated in colorectalcarcinomas. Overexpression of p53 is frequently noted in colorectal carcinomas with deep invasion or lymph node metastasis. However, there have been few reports about the association between MUC1, MUC2, and p53 expression with respect to the metastatic potential. This study was aimed to investigate the relationship of MUC1, MUC2, and protein p53 expressions with clinicopathological factors in colorectal carcinomas. Expressions of MUC1, MUC2, and p53 protein were examined immunohistochemically. Of total 97 cancers, 44 (45%) were MUC1 positive, 39 (40%) were MUC2 positive and 58 (59%) showed a p53 overexpression. Coexpression of MUC1 with p53 and dual expression of MUC1 with MUC2 were associated with a higher frequency of lymph node metastasis (p<0.05). The right colon showed a higher MUC1 positivity and frequent lymph node metastasis than the left colon (p<0.05). These results suggest that the coexpression of MUC 1 with p53 or MUC2 are involved in regional lymph node metastasis in colorectal carcinomas. The high expression of MUC1 in the right colon cancer was revealed to relate with lymph node metastasis.     

bcl-2 protein expression is associated with better prognosis in colorectal cancer

AIMS: To investigate the expression of bcl-2 in colorectal carcinoma and to examine its association with mediators of apoptosis (p53 and mdm-2), clinicopathological features and long-term outcome. METHODS AND RESULTS: We determined by immunohistochemistry the expression of bcl-2 in 102 colorectal carcinomas with 10-year follow-up. In 66 of these cases in which we had previously assessed p53 status, no correlation was seen between bcl-2 and p53. The mdm-2 protein was not detected in any of the 66 cases. Cytoplasmic staining of the bcl-2 gene product was seen in the tumour cells of 22 cases (22%). Using a polymerase chain reaction technique we showed that overexpression of bcl-2 was not due to rearrangement of the bcl-2 gene. Expression of bcl-2 protein was related to tumour grade but was unrelated to patient age, sex, tumour site, tumour size or Dukes' stage. There was a trend towards increased survival in those whose tumours expressed bcl-2 protein (P = 0.055). When entered into a multivariate analysis, this survival difference was independent of tumour stage (P = 0.05). CONCLUSIONS: These results suggest that bcl-2 expression in colorectal carcinoma is associated with a better long-term prognosis.     

结直肠肿瘤P53蛋白表达与肿瘤临床病理学特征及预后的关系

应用免疫组织化学方法观察７２例结直肠、１１例腺瘤、３０例癌旁粘主１５例正常粘膜Ｐ５３蛋白表达及其与肿瘤临床病理学特征和预后的关系。结果显示：结直肠癌Ｐ５３蛋白阳性率为５０％，腺瘤的阳性率为１８．１８％（Ｐ＜０．０５），阳性细胞主要分布在腺瘤的增生区或不典型增生区；正常粘膜、癌粘膜Ｐ５３蛋白均阴性。Ｐ５３蛋白阳性的结直肠癌多呈浸润性生长方式，且以浸润至浆膜外者居多，患者片存率较Ｐ５３蛋白阴性者（Ｐ＜     

p53和bcl-2蛋白过度表达与大肠癌生物学行为的关系

目的：探讨p53蛋白和bcl－2蛋白共同表达与大肠癌生物学行为的关系。方法：应用免疫组织化学染色ABC法检测p53蛋白及bcl－2蛋白在67例大肠癌组织中的表达。结果：全阴性组和p53阴性bcl－2阳性组的PCNA增殖指数低于p53阳性bcl－2阴性组及全阳性组（P＜0．01或P＜0．05）。全阳性、p53阳性bcl－2阴性组及p53阴性bcl－2阳性组均多呈浸润性生长（P＜0．01或P＜0．05）。全阳组浸润深度多至浆膜外（P＜0．01或P＜0．05）。两蛋白全阴性组5年生存率高（P＜0．05）。P53和bcl－2蛋白表达与大肠癌Dukes分期、淋巴结转移和组织学类型均无统计学意义。结论：bcl－2蛋白表达对细胞增殖的相关性不大。主要是p53蛋白的作用。只要有1种蛋白表达时大肠癌即多呈浸润性生长，两种蛋白全部阳性组浸润深度较深，蛋白全部阴性组的预后较好，提示p53和bcl－2蛋白的表达情况可部分地反映大肠癌的生物学行为。     

Clinicopathological significance and survival influence of p53 protein expression in gastric carcinoma

AIMS: Mutations in the gene coding for p53 protein are among the most frequent genetic alterations observed in human cancers. The relevance and biological significance of p53 expression in gastric carcinoma are far from being fully established. The aim of our study was to evaluate the influence of p53 detected by immunohistochemistry in the clinicopathological behaviour of a series of gastric carcinoma cases. METHODS AND RESULTS: Samples from 163 patients treated by gastric resection for gastric carcinoma between 1988 and 1995 were used. Surgical specimens were evaluated for the presence of p53 protein detected by immunohistochemistry with a monoclonal antibody. Cases were classified as positive or negative for p53. Several clinicopathological parameters and c-erb B-2 expression were analysed in the same series and compared with the expression of p53. Cumulative survival was evaluated using univariate analysis and Cox model regression. p53 expression was identified in 41 carcinomas (25.2%) and was significantly associated with venous invasion (P = 0.049), lymph node metastases (P = 0.01) and c-erb B-2 expression (P = 0.003). All the parameters except gender, tumour size and Laurén's classification influenced survival on univariate analysis. p53 expression correlated with overall survival (P = 0.006) and survival in the subgroup of patients with intestinal type carcinoma (P = 0.04). In the subgroup of patients with carcinomas not expressing c-erb B-2, p53 expression significantly influenced cumulative survival (P = 0.02). CONCLUSIONS: p53 expression is associated with the aggressive biological behaviour of gastric carcinomas and is related to cumulative survival.     

EARLY EXPRESSION OF bcl-2 PROTEIN IN THE ADENOMA–CARCINOMA SEQUENCE OF COLORECTAL NEOPLASIA

bcl-2 was originally identified as an oncogene involved in follicular lymphomas as a result of chromosomal translocation (14;18). It is now believed that bcl-2 is implicated in the regulation of cell death by inhibiting apoptosis and that its expression is not restricted to haematopoietic cells, but is also present in many epithelia and mesenchymal tissues. Recent studies have analysed the expression of this molecule in a variety of non-lymphoid malignancies including lung, breast, prostate, and nasopharyngeal carcinomas and neuroblastoma. In the present study, 50 colorectal adenomas, 10 hyperplastic polyps, and 142 carcinomas, including 25 arising from pre-existing adenomas, were examined using an antibody detecting the bcl-2 protein product. In non-neoplastic mucosa, bcl-2 was expressed in the crypt cells only, whilst the more differentiated surface epithelial cells lacked any demonstrable bcl-2. Forty-one of the 50 adenomas (82 per cent) and 48 of the 142 carcinomas were positive for bcl-2 expression. All hyperplastic polyps were negative. A reciprocal relationship was found between bcl-2 reactivity and p53 overexpression, as detected by DO7 antibody, in approximately 65 per cent of the cases. The bcl-2-positive/p53-negative subgroup showed a strong correlation ( P =0·0056) with negative lymph node status (Dukes' A and B), implying a less aggressive pathway of neoplastic transformation.     

宫颈癌中COX-2与p53、E-cadherin蛋白表达的关系

目的　探讨宫颈癌组织中环氧合酶 2 (COX 2 )的表达与 p5 3、E cadherin蛋白表达的关系。 方法　采用免疫组织化学S P法检测 4 1例宫颈癌和 10例正常宫颈上皮组织中COX 2、p5 3、E cadherin蛋白的表达水平。结果　宫颈癌组织中COX 2、p5 3表达水平明显高于正常宫颈上皮 (P <0 .0 1) ,而E cadherin蛋白的表达水平明显低于正常宫颈上皮 (P <0 .0 1) ;COX 2的高表达与宫颈癌淋巴结转移和浸润深度有关 (P <0 .0 5 ) ,与患者年龄、临床分期、组织学类型、分化程度无关 (P >0 0 5 ) ;COX 2表达阳性组 p5 3的表达水平明显高于COX 2表达阴性组 (P <0 0 5 ) ,COX 2表达阳性组的E cadherin蛋白的表达水平则低于相对应阴性组 ,差异有显著性 (P <0 0 5 )。结论　在宫颈癌的发生发展中COX 2、p5 3、E cadherin可能起重要作用。COX 2通过使抑癌基因p5 3失活 ,降低细胞黏附分子E cadherin的水平促进宫颈癌的发生。     

胃癌组织中p27KIP1和MMP-9的表达及其临床意义

目的：探讨p^27KIP和MMP－9在胃癌中的表达其与临床病理参数之间的关系。方法：采用免疫组化S－P法检测56例胃癌中p^27KIP和MMP－9蛋白的表达。结果：p^27KIP和MMP－9蛋白在胃癌中的阳性率分别为46．4％和53．6％。p^27KIP蛋白的表达与肿瘤的分化程度、Lauren分型、淋巴结转移和浸润深度有关（P＜0．05），MMP－9的表达只与胃癌的Lauren分型、淋巴结转移和浸润深度有关（P＜0．05），而与分化程度无关（P＞0．05）。结论：p^27KIP蛋白低表达参与了胃癌的发展，MMP－9可能在胃癌的侵袭和转移中发挥作用。     

Prognostic evaluation of metallothionein expression in human colorectal neoplasms

AIM: To investigate the role of metallothionein in colorectal tumours and the possible relation with other factors associated with tumour progression: expression of cathepsin D (CD), CD44, p53, Rb, bcl-2, c-erbB-2, epidermal growth factor receptor (EGFR), proliferation indices (Ki-67, proliferating cell nuclear antigen (PCNA)), and conventional clinicopathological variables. METHODS: The immunohistochemical expression of metallothionein was investigated in 23 cases of colorectal adenoma and 94 adenocarcinomas. Metallothionein expression was examined by the avidinbiotin peroxidase immunoperoxidase (ABC) using the monoclonal mouse antibody E9, on formalin fixed, paraffin embedded tissue. RESULTS: Positive metallothionein expression (> 5% of neoplastic cells) was observed in 30.4% of adenomas and 25.5% of adenocarcinomas, while 8.7% of adenomas and 14.9% carcinomas showed focal metallothionein positivity. In contrast, 60.9% of adenomas and 59.6% of carcinomas almost completely lacked metallothionein expression. In the series of adenocarcinomas, metallothionein expression was inversely correlated with CD44 in neoplastic cells (p = 0.01). There was no statistically significant difference of metallothionein expression, or the other variables examined, between adenocarcinomas and adenomas. CONCLUSIONS: Metallothionein expression does not seem to indicate aggressive biological behaviour in colorectal adenocarcinomas, in comparison with the other types of carcinoma. The inverse correlation with CD44 could suggest that the decreased metallothionein expression may contribute to the metastatic spread of the lymph node involvement in colorectal cancer. Metallothionein expression does not seem to represent an independent prognostic marker in colorectal cancer.