抑癌基因APC蛋白在结直肠粘膜及结直肠癌中的表达

众多研究表明位于染色体５ｑ上抑癌基因ＡＰＣ的突变、缺失可能与结直肠癌的发生发展相关，我们应用免疫组化技术分析了ＡＰＣ蛋白在正常结直肠粘膜的表达特点以及其表达异常与结直肠癌生物学行为的关系。一、材料和方法共收集我院１９９３年１月至１９９４年１０月间结直肠癌手术标本４５例，每例包括原发癌肿和手术切缘正常粘膜。用中性甲醛溶液固定、石蜡包埋，制成５μｍ厚切片做ＨＥ及免疫组化染色。ＡＰＣ兔抗人多克隆抗体Ｃ２０（１∶１００）为ＳａｎｔａＣｒｕｚ公司产品，ＳＰ试剂盒购于福州迈新公司，染色采用ＳＰ法，微波修复…     

结直肠癌中高尔基体蛋白73的表达及意义

目的 探讨结直肠癌中高尔基体蛋白73(golgin protrin 73, GP73)的表达,及其联合常见肿瘤指标CEA、CA125、CA19-9在结直肠癌中的诊断价值。方法 收集50例结直肠癌和50例健康体检者的临床资料以及石蜡标本。相关性分析GP73和CEA、CA125、CA19-9在结直肠癌和正常对照之间的关系,及血清GP73与结直肠癌其它肿瘤指标、病理特征和组织中GP73表达的关系,进一步通过免疫组化法分析CEA和GP73在结直肠癌和癌旁正常组织中的表达情况及两者的相关性。结果 血清指标CEA、GP73在结直肠癌和正常对照组组间比较差异有统计学意义(P<0.001);相关性分析显示：血清GP73与结直肠癌病理分期和组织中GP73表达具有相关性(P<0.05);GP73分别联合CEA、CA125、CA19-9诊断分析显示：CEA+GP73能明显提高结直肠癌的诊断价值(AUC=0.815,P<0.05)。免疫组化结果显示：CEA在结直肠癌组织中的高表达率为80%,明显高于癌旁正常组织的高表达率24%(P<0.05);GP73在结直肠癌组织中的高表达率为76%...     

结直肠癌组织中CD151表达及其临床意义

目的 探讨结直肠癌组织中CD151的表达及其与结直肠癌发生、发展和转移的关系.方法 采用实时荧光定量PCR技术检测22例肠癌组织及癌旁组织中CD151mRNA的表达,采用免疫组化SP法检测此22例及另67例石蜡包埋肠癌组织及癌旁组织中CD151蛋白的表达,并分析其与肠癌临床病理因素之间的关系.结果 CD151mRNA和蛋白在结直肠癌中的表达明显高于癌旁组织,其蛋白的表达与结直肠癌浸润深度、淋巴结转移和TNM分期密切相关,差异具有统计学意义(P<0.05).结论 CD151可望成为与结直肠癌浸润、转移有关并提示预后不良的指标.     

结直肠癌中Survivin和P-gp蛋白表达及其与血清CEA的相关性

目的探讨结直肠癌组织中Survivin和P-gp蛋白表达及其与血清CEA的相关性。方法采用免疫组化SP法检测63例结直肠癌及癌旁黏膜,10例腺瘤组织中Survivin和P-gp蛋白表达,分析两者表达与结直肠癌临床病理特征以及血清CEA值的关系。结果 Survivin和P-gp蛋白在结直肠癌组织中阳性率分别为74.6%、76.19%,均显著高于癌旁和腺瘤组织(P 0.01)。Survivin和P-gp蛋白表达与结直肠癌淋巴结转移、远处转移、TNM分期呈正相关(P 0.05),两者表达之间具有正相关性(P 0.05),Survivin蛋白高表达组血清CEA值明显高于低表达组(P 0.05)。结论 Survivin和P-gp蛋白在结直肠癌中高表达与肿瘤细胞的侵袭、转移密切相关,两者的表达具有协同作用,联合血清CEA检测有助于对肿瘤监测,为治疗结直肠癌提供分子生物学支持。     

结直肠癌转移动物模型血清中几种蛋白变化的观察

目的建立可视化结直肠癌转移模型,并应用血清蛋白质组学技术分析可视化结直肠癌转移模型血清蛋白质的变化。方法用pEGFP—N1绿色荧光蛋白基因转染具有转移能力的SW480细胞株,在裸鼠皮下接种成瘤后,再原位接种于裸鼠右半结肠,利用整体荧光成像系统观察结直肠癌原位成瘤及发生转移的情况。同时采集裸鼠原位成瘤后不同转移阶段的血清,进行双向电泳联合时间飞行质谱分析差异蛋白质,观察结直肠癌转移模型裸鼠癌转移前后血清蛋白的变化。结果获得稳定表达绿色荧光蛋白的SW480-EGFP细胞株,裸鼠皮下接种成瘤率为100％,原位种植成瘤率为10／10,区域淋巴结转移为10／10,肝转移率为4／10,肺转移率为3／10。通过血清蛋白质组学技术成功鉴定了5个在癌转移发生后血清内显著升高的血清蛋白：结合珠蛋白α链,载脂蛋白A4,载脂蛋白E,免疫球蛋白κ型V区L链和转铁蛋白。结论成功建立结直肠癌可视化转移模型,利用不同转移阶段鼠血清进行双向电泳图谱差异分析筛选出5个在癌转移发生后血清中显著升高的蛋白可能是与结直肠癌转移相关的蛋白。     

结直肠癌组织中CD10的表达及其临床意义

目的探讨结直肠癌组织中间质和上皮细胞CD10的表达及与肿瘤临床生物学行为的关系。方法运用免疫组化EnVision法检测90例结直肠癌组织和20例正常黏膜中CD10的表达,同时用α-SMA标记结直肠癌组织中肌纤维母细胞。结果正常结直肠黏膜中间质和上皮细胞均无CD10的表达,而在结直肠癌组织中,CD10在间质和上皮细胞的阳性表达率分别为57．8％和26．7％。结直肠癌中CD10阳性的间质细胞和上皮细胞在高、低级别癌中的表达差异无显著性（P〉0．05）;间质细胞CD10阳性在侵及肌层者高于侵及浆膜层者（P〈0．01）,而CD10阳性的上皮细胞与肿瘤的浸润深度无关（P〉0．05）;间质细胞和上皮细胞表达CD10与淋巴结转移均无相关性（P〉0．05）;CD10阳性间质细胞在有脉管侵犯者低于无脉管侵犯者（P〈0．01）,CD10阳性上皮细胞在有脉管侵犯者高于无脉管侵犯者（P〈0．01）。CD10阳性间质细胞与α-SMA阳性间质细胞分布一致。结论结直肠癌中间质细胞肌纤维母细胞化可阻止肿瘤的转移,而肿瘤细胞表达CD10可能通过促进细胞外基质溶解而促进转移。     

结直肠癌mdm2,p21和p53蛋白的表达

一、材料与方法1.标本 :89份石蜡组织取自 1991～ 1994年期间本院结直肠腺癌手术切除标本 ,术后随访 3～ 6年。其中高分化腺癌 30例 ,中分化腺癌 44例 ,低分化腺癌 15例。按Ｄｕｋｅｓ临床分期 ,无Ａ期病例 ,Ｂ期 42例 ,Ｃ期 37例 ,Ｄ期 10例。纤维结肠镜及病理检查未发现异常的结直肠粘膜活检标本 12例做对照。组织均常规甲醛溶液固定 ,石蜡包埋 ,连续切片 ,分别作常规ＨＥ染色 ,ｍｄｍ2、ｐ2 1、ｐ5 3蛋白免疫组化染色。2 .免疫组化染色 :采用ＳＰ法 ,鼠抗ｍｄｍ2 (ＳＭｐ14)、ｐ5 3(Ｄｏ 1)、ｐ2 1(187)为ＳａｎｔａＣｒｕｚ公司试剂 ,Ｓ…     

ERK5在结直肠癌组织中的表达及其与结直肠癌转移的关系

目的: 检测和分析细胞外信号调节激酶5(ERK5)蛋白在原发性结直肠癌组织和癌旁正常黏膜组织中的表达情况及其与结直肠癌临床病理参数的关系,探讨ERK5的临床意义。方法: 使用组织芯片和免疫组化的方法检测338例原发性结直肠癌癌组织和80例癌旁正常组织中ERK5的表达,分析ERK5表达与结直肠癌的临床病理参数及预后的相关性。结果:80例癌旁正常黏膜中,ERK5低表达59例(73.8%)、高表达为21例(26.2%);338例结直肠癌组织中ERK5低表达204例(60.4%)、高表达134例(39.6%);二者中的ERK5蛋白表达有显著差异(P<0.05)。在338例结直肠癌组织中,27例发生远处转移患者的癌组织中ERK5高表达率为63.0%,而311例未发生远处转移患者的癌组织中ERK5高表达率为37.6%,二者之间有显著差异(P<0.05)。但癌组织中ERK5的表达水平与患者性别、年龄、体重指数、肿瘤位置、T分期、N分期、TNM分期和分化程度无明显相关性(P>0.05)。Kaplan-Meier生存分析显示在结直肠癌组织中ERK5高表达的病人总体5年生存率与低表达组无明显差异(P>0.05)。结论:ERK5蛋白在癌组织中的表达较癌旁正常组织高;发生远处转移的结直肠癌组织中ERK5蛋白的表达水平明显高于未发生远处转移的癌组织中ERK5水平。ERK5可能是一个促进肿瘤远处转移的因子。     

Tiam1对结直肠癌细胞生物学特性的影响

目的探讨Tiam1(Tlymphomainvasionandmetastasis)对结直肠癌细胞株生物学特性的影响。方法用逆转录聚合酶链反应方法检测Tiam1基因在结直肠癌细胞株中的表达,筛选Tiam1不表达的细胞株;将Tiam1/C1199HAcDNA导入内源性不表达Tiam1基因的人结直肠癌HT29细胞株中,G418筛选抗性克隆,免疫组织化学和Western蛋白印迹法鉴定转染后Tiam1基因在细胞中的表达,利用四甲基偶氮唑盐(MTT)法检测Tiam1对细胞增殖的影响,体外侵袭实验检测Tiam1对结直肠癌侵袭转移的影响。结果Tiam1基因在高转移的LoVo和SW620中高表达,在低转移的HT29中不表达,在SW480和HCT116细胞表达相应较低。通过7d的连续比较,HT29/mock和HT29/Tiam1两组的细胞增殖能力差异有统计学意义(F=10.512,P=0.003)。Tiam1基因的导入使结直肠癌细胞的增殖能力明显增强,体外侵袭转移能力显著增加,HT29/Tiam1穿过细胞为(88.6±9.2)个/200倍视野,HT29/mock为(46.8±3.4)个/200倍视野,二者的差异具有统计学意义(t=9.54,P<0.01)。结论Tiam1基因具有促进结直肠癌细胞增殖的能力,Tiam1与结直肠癌的侵袭转移密切相关,Tiam1表达可作为结直肠癌侵袭转移过程中一个有价值的指标。     

结直肠癌原发灶及淋巴结转移灶中SATB2表达及意义

目的：探讨富含AT序列特异性结合蛋白2(special AT-rich sequence-binding protein 2, SATB2)在结直肠癌组织及对应淋巴结转移灶中的表达,并分析其与临床病理特征的关系。方法应用免疫组化EnVision法检测200例结直肠癌原发灶及80例淋巴结转移灶中SATB2蛋白表达。结果结直肠癌原发灶中SATB2高表达率为25．0%(50/200),中等强度表达率为36．0%(72/200),阴性率为39．0%(78/200)。对应淋巴结转移灶中SATB2高表达率为15．0%(12/80),中等强度表达率为28．8%(23/80),阴性率为56．2%(45/80)。 SATB2在结直肠癌原发灶中的表达与肿瘤大小、分化程度,浸润深度、淋巴结转移以及TNM分期显著相关(P<0．05),与患者年龄、性别及远处转移无相关性。 SATB2在结直肠癌淋巴结转移灶中表达显著低于原发灶(P<0．05),其在转移灶中的表达与临床病理因素无明显相关性。结论 SATB2低表达与结直肠癌的发生、发展相关,在肿瘤转移过程中表达明显降低,有望成为新型的结直肠癌分子靶标。     

Overexpression of serine-threonine receptor kinase-associated protein in colorectal cancers

Transforming growth factor-beta (TGF-beta) regulates many cellular processes, including cellular proliferation and differentiation. Disruption of the TGF-beta signaling pathway can lead to cancer. Serine-threonine receptor kinase-associated protein (STRAP), an inhibitor of TGF-beta signaling, is an important regulator of cell proliferation. Here, in order to investigate the roles of STRAP in colorectal carcinogenesis, the expression of the STRAP protein was investigated in 59 colonic adenomas and 123 colorectal cancers by immunohistochemistry. Upregulation of STRAP protein was observed in 30 (50.8%) of 59 adenomas and 87 (70.7%) of 123 cancers, respectively. Statistically, overexpression of STRAP protein was not associated with clinicopathological parameters and 5 year survival (P > 0.05). Interestingly, significant association was observed between STRAP and Ki-67 positivity (P < 0.05), suggesting that STRAP contributes to an increased proliferate potential of tumor cells. These results indicate that upregulation of STRAP might play a role in tumor development as an early event for colorectal cancers.     

Overexpression of carbonic anhydrase-related protein VIII in human colorectal cancer

Carbonic anhydrase-related protein (CA-RP) VIII, which is a member of the CA gene family, has been shown to have no catalytic CA activity and its biological function is still unknown. Recently, overexpression of CAs IX and XII has been reported in certain types of malignancy. To investigate a potential role for CA-RP VIII in human colorectal epithelial carcinogenesis, colorectal tissue specimens from surgically resected adenocarcinomas (n = 60) and endoscopically polypectomized adenomas (n = 13) were analysed by immunohistochemistry using monoclonal antibodies to CA-RP VIII and Ki-67 antigen. Less than 5% of epithelial cells in normal colonic mucosae (n = 73) were CA-RP VIII-positive and these were localized to the deep part of the cryptal epithelium. Increased expression of CA-RP VIII was observed in 78% of colorectal carcinomas. An intense signal was frequently observed at the tumour invasion front and its distribution was completely different from that of Ki-67 antigen. Colorectal adenomas also showed significant immunopositivity for CA-RP VIII, but its expression level was much lower than in adenocarcinomas. These findings suggest that CA-RP VIII plays a role in the process of invasion in colorectal cancer.     

The mRNA and protein expression of A-kinase anchor proteins 13 in human colorectal cancer

There was no literature reporting the relationship between AKAP13 and colorectal carcinoma. This study is aim to investigate the expression and role of AKAP13 in human colorectal cancers. This study investigated 94 pair-matched human colorectal cancers and adjacent normal mucosa, as well as 36 adenomas, of which mRNA expression of AKAP13 was detected by relative Quantitative-Real-Time RT-PCR and protein expression by immunohistochemical staining. AKAP13 gene was upregulated in colorectal cancer group by 2.259 times compared to control group without significant difference (P = 0.081), and no expression was detected in adenoma by RT-PCR. The positive expression rate of AKAP13 protein in colorectal carcinoma (52.3%) was significantly higher than those in adenoma (9.1%) and normal tissue (34.7%) (P = 0.006) by immunohistochemical staining. Either the mRNA or protein expressions of AKAP13 were correlated with histological types and differentiation grade (P < 0.05). Our results suggest AKAP13 protein may be related to the carcinogenesis of human colorectal cancer. However, more deeply and larger scale research are required to prove the correlation.     

Overexpression of EIF-5A2 is associated with metastasis of human colorectal carcinoma

Our previous study has suggested an oncogenic role of eIF-5A2 in ovarian tumorigenesis. Abnormalities of eIF-5A2, however, in colorectal carcinoma are unclear. In this study, amplification and overexpression of eIF-5A2 in colorectal carcinoma were studied by fluorescence in situ hybridization and immunohistochemistry using colorectal carcinoma tissue microarrays, including 139 primary colorectal carcinomas and their adjacent normal mucosa, 22 paired premalignant adenomas, and 42 metastatic tumors. The immunohistochemistry results showed that overexpression of EIF-5A2 was detected in none of normal epithelial mucosa, 35.3% of colorectal adenomas, 53.2% of primary colorectal carcinomas, and 67.6% of metastases. Amplification of eIF-5A2 was detected in 15.8% (16/101) of informative colorectal carcinomas, and most of them showed overexpression of EIF-5A2. In primary colorectal carcinomas, the frequency of EIF-5A2 overexpression was significantly higher in colorectal carcinomas with lymphovascular invasion (61.2%) than that in colorectal carcinomas without lymphovascular invasion (36.6%, P < .05). In addition, significant positive associations were found between EIF-5A2 overexpression and the tumors' later pN and pM stages, as well as increased tumor cell proliferation (P < .05). These findings suggest that overexpression of EIF-5A2 in colorectal carcinomas may be important in the acquisition of a metastatic phenotype and plays an important role in colorectal carcinoma development and progression.     

结直肠癌中p-AKT和PTEN蛋白的表达及其临床意义

目的探讨磷酸化AKT(p-AKT)和PTEN蛋白在结直肠癌中的表达及其临床意义。方法应用免疫组化PV-9000两步法检测80例结直肠癌手术切除标本和20例癌旁正常组织中p-AKT、PTEN蛋白的表达,分析两者与临床病理特征的关系。结果 p-AKT蛋白在结直肠癌中表达的阳性率(71.25%)显著高于癌旁正常组织(10.00%,P0.05);而PTEN蛋白在结直肠癌中表达的阳性率(45.00%)与癌旁正常组织(100.00%,P0.05)相比具有统计学意义的降低,二者的表达与肿瘤的分化程度、TNM分期、浆膜浸润、淋巴结转移具有明显相关性(P0.05),并且p-AKT与PTEN蛋白的表达呈显著负相关(r=-0.314,P=0.005)。结论 p-AKT和PTEN蛋白的表达可能与结直肠癌的发生、发展密切相关。联合检测二者的表达,对判断结直肠癌的恶性程度及预后具有重要的临床意义。     

Overexpression of p53 protein and Ki67 proliferative index in hepatocellular carcinoma:

The overexpression of p53 protein and the Ki67 proliferative index was evaluated in 96 hepatocellular carcinomas (HCC), 67 in cirrhotic livers and 29 in non-cirrhotic ones, and in 13 non-carcinomatous lesions, all surgically resected from Italian patients. Overexpression of p53 was detected only in carcinomatous lesions, and was significantly related to the grade of HCC ( P < 0.001). In fact, p53 was observed in 7/7 (100%) cases of grade IV, 13/43 (30.3%) of grade III, and 10/ 46 (21.7%) of grade II. The relationship between p53 and Ki67 scores was determined in serial sections from corresponding areas of both diffuse and patchy immunoreactivity. In the overall population, p53-positive tumors showed a significantly higher Ki67 score (15.9 ± 5.5% vs 9.2 ± 4.3% [ P < 0.001]). This observation was evident in all grades of HCC.     

c-erbB-2过表达、nm23低表达与结直肠癌预后不良的关系

目的　探讨结直肠癌c erbB 2、CD4 4v6、nm2 3基因蛋白表达与临床病理特征的相关性及意义。方法　用免疫组织化学SP法检测了 92例结直肠癌组织c erbB 2、CD4 4v6、nm2 3蛋白表达 ,并对其中随访 10年以上的 2 8例作了生存分析。结果　c erbB 2和CD4 4v6的表达均与国际抗癌联盟(UICC)分期有关 (P <0 0 5 ) ,而且c erbB 2表达还与显著的瘤周淋巴样细胞浸润有关 (P <0 0 5 )。在随访 10年以上的 2 8例中 ,单因素分析显示 :患者生存率与组织分型、UICC分期、肿瘤生长方式、瘤周淋巴样细胞浸润以及c erbB 2、CD4 4v6、nm2 3表达均有关。但经多因素Cox比例风险分析 ,只有UICC分期、c erbB 2和nm2 3表达具有独立的预后意义。结论　除UICCⅢ、Ⅳ期之外 ,c erbB 2过表达和nm2 3表达减少似乎可作为结直肠癌预后不良的可行性指标