High level of ezrin expression in colorectal cancer tissues is closely related to tumor malignancy

AIM： To investigate the ezrin expression in normal colorectal mucosa and colorectal cancer tissues, and study the correlation between ezrin expression in colorectal cancer tissues and tumor invasion and metastasis. METHODS： Eighty paraffin-embedded cancer tissue samples were selected from primary colorectal adenocarcinoma. Twenty-eight patients had well- differentiated, 22 had moderately differentiated and 30 had poorly differentiated adenocarcinoma. Forty-five patients and 35 patients had lymph node metastasis. Forty-five patients were of Dukes A to B stage, and 35 were of C to D stage. Another 22 paraffin-embedded tissue blocks of normal colorectal epithelium （〉 5 cm away from the edge of the tumor） were selected as the control group. All patients with colorectal cancer were treated surgically and diagnosed histologically, without preoperative chemotherapy or radiotherapy. The immunohistochemistry was used to detect the ezrin expression in paraffin-embedded normal colorectal mucosa tissues and colorectal cancer tissue samples. RESULTS： Ezrin expression in colorectal cancer was significantly higher than in normal colorectal mucosa （75.00% vs 9.09%, P 〈 0.01）, and there was a close relationship between ezrin expression and the degree of tumor differentiation, lymph node metastasis and Dukes stage （88.46% vs 50.00%, P 〈 0.01; 94.28% vs 51.11%, P 〈 0.01; 94.28% vs 51.11%, P 〈 0.01）. CONCLUSION： Ezrin expression is obviously higher in colorectal cancer tissues than in normal colorectal mucosa tissues, and the high level of ezrin expression is closely related to the colorectal cancer invasion and metastasis process.     

Mad2 and p53 expression profiles in colorectal cancer and its clinical significance

AIM: To investigate the expression of tumor suppressor gene p53 and spindle checkpoint gene Mad2, and to demonstrate their expression difference in colorectal cancer and normal mucosa and to evaluate its clinical significance.METHODS: Westemn blot and immunohistochemistry methods were used to analyze the expression of Mad2 in colorectal cancer and its corresponding normal mucosa. The expression of p53 was detected by immunohistochemistry method in colorectal cancer and its corresponding normal mucosa.RESULTS: Mad2 was significantly overexpressed in colorectal cancer compared with corresponding normal mucosa (P＜0.001), and it was not related to the differentiation of adenocarcinoma and other dinical factors (P＞0.05).The ratio of Mad2 protein in cancer tissue (C) to that in its normal mucosa tissue (N) was higher than 2, which was more frequently observed in patients with lymph gland metastasis (P＜0.05). p53 protein expression was not observed in normal mucosa. The rate of p53 positive expression in adenocarcinomas was 52.6%. There was a significant difference between adenocarcinomas and normal mucosa(P＜0.001), which was not related to the differentiation degree of adenocarcinoma and other clinical factors (P＞0.05).CONCLUSION: Defect of spindle checkpoint gene Mad2and mutation of p53 gene are involved mainly in colorectal carcinogenesis and C/N＞2 is associated with prognosis of colorectal cancer.     

Oncogenic role of clusterin overexpression in multistage colorectal tumorigenesis and progression

AIM: To investigate the expression pattern of clusterin in colorectal adenoma-carcinoma-metastasis series,and to explore the potential role of clusterin in multistage colorectal tumorigenesis and progression. METHODS: A colorectal carcinoma (CRC)-tissue microarray (TMA), which contained 85 advanced CRCs including 43 cases of Dukes B,21 of Dukes C and 21 of Dukes D tumors, were used for assessing the expression of clusterin (clone 41D) and tumor cell apoptotic index (AI) by immunohist-ochemistry and TUNEL assay,respectively.Moreover the potential correlation of dusterin expression with the patient's clinical-pathological features were also examined. RESULTS: The positive staining of clusterin in different colorectal tissues was primarily a cytoplasmic pattern. Cytoplasmic overexpression of clusterin was detected in none of the normal colorectal mucosa, 17% of the adenomas, 46% of the primary CRCs, and 57% of the CRC metastatic lesions. In addition, a significant positive correlation between overexpression of clusterin and advanced clinical (Dukes) stage was observed (P<0.01).Overexpression of cytoplasmic clusterin in CRCs was inversely correlated with tumor apoptotic index (P<0.01),indicating the anti-apoptotic function of cytoplasmic clusterin in CRCs. CONCLUSION:These data suggests that overexpression of cytoplasmic dusterin might be involved in the tumorigenesis and/or progression of CRCs. The anti-apoptotic function of cytoplasmic dusterin may be responsible, at least in part, for the development and biologically aggressive behavior of CRC.     

KISS1 methylation and expression as predictors of disease progression in colorectal cancer patients

AIM:To examine the effect of aberrant methylation of the KISS1 promoter on the development of colorectal cancer(CRC)and to investigate reversing aberrant methylation of the KISS1 promoter as a potential therapeutic target.METHODS:KISS1 promoter methylation,mRNA expression and protein expression were detected by methylation-specific polymerase chain reaction(PCR),real-time quantitative PCR and Western blotting,respectively,in 126 CRC tissues and 142 normal colorectal tissues.Human CRC cells with KISS1 promoter hypermethylation and poor KISS1 expression were treated in vitro with 5-aza-2’-deoxycytidine(5-Aza-CdR).After treatment,KISS1 promoter methylation,KISS1 mRNA and protein expression and cell migration and invasion were evaluated.RESULTS:Hypermethylation of KISS1 occurred frequently in CRC samples(83.1%,105/126),but was infrequent in normal colorectal tissues(6.34%,9/142).Moreover,KISS1 methylation was associated with tumor differentiation,the depth of invasion,lymph node metastasis and distant metastasis(P<0.001).KISS1methylation was also associated with low KISS1 expression(P<0.001).Furthermore,we observed re-expression of the KISS1 gene and decreased cell migration after 5-Aza-CdR treatment in a CRC cell line.CONCLUSION:These data suggest that KISS1 is down-regulated in cancer tissues via promoter hypermethylation and therefore may represent a candidate target for treating metastatic CRC.     

Relationship between somatostatin receptor subtype expression and clinicopathology, Ki-67, Bcl-2 and p53 in colorectal cancer

AIM: To study the SSTR1, 2, 3, 4, 5 expression and their relationships with clinico-pathological factors, cell proliferation, Bcl-2 and p53 expression in colorectal cancer cells. METHODS: Immunohistochemical staining of five SSTR subtypes, Ki-67, Bcl-2 and p53 was performed by the standard streptavidin-peroxidase (SP) technique for the paraffin sections of 127 colorectal cancers, and expression of five SSTR subtypes in 40 specimens of normal colorectal mucosae was detected with the same method. RESULTS: Positive staining for five SSTR subtypes was observed in colorectal cancer cells and normal colorectal mucosae. SSTR1 was the most predominant subtype in both colorectal cancer and normal colorectal mucosa, and the second was SSTR5 or SSTR2. As compared with normal colorectal mucosa, SSTR4 was more frequently expressed in colorectal cancer cells (2.5% vs 18.9%, P< 0.05); the expression of SSTR2, 4, 5 in moderately to well differentiated colorectal adenocarcinoma was significantly higher than that in poorly differentiated ones (P< 0.05), the SSTR1 expression in colorectal cancer with positive lymph node metastasis was significantly higher than that with negative lymph node metastasis (72.2% and 54.5%, P< 0.05). In addition, in the ulcerative type of colorectal cancer, SSTR2 expression was obviously decreased (P < 0.05); the correlation did not reach a statistical significance between the five SSTR subtypes expression and Dukes'stages (P> 0.05), but the frequency of SSTR1 expression increased with Dukes' stage, while SSTR3 and SSTR5 expression decreased with Dukes' stage. Moreover, there was no correlation between expression of the five SSTR subtypes and other clinicopathological factors such as age, sex, tumor site, tumor depth, distant metastasis. The proliferative indexes in colorectal cancer cells with negative expression of SSTR2 and SSTR3 were significantly higher than that with positive expression (P<0.05). The Bcl-2 expression in colorectal cancer cells with positive expression of SSTRl, 2, 3, 5 was significantly lower than that with negative expression (P<0.05). There was no correlation between five SSTR subtypes and p53 expression. CONCLUSION: The most predominant SSTR subtype is SSTR1, and the second is SSTR2 or SSTR5. Five SSTR subtypes play different roles in the development of colorectal cancer. SSTR2 and SSTR3 can inhibit the proliferation and promote apoptosis of tumor cells.     

KAI1 gene expression in colonic carcinoma and its clinical significances

AIM:To investigate KAI1 gene expression in the progressionof human colonic carcinoma and its clinical significances.METHODS:KAI1 expression was detected by in situhybridization and immunohistochemistry in the 4 establishedcell lines of colorectal carcinoma with different metastaticpotentials,and in 80 specimens of colonic carcinoma,21colonic carcinoma specimens with lymphatic metastasisand 20 controls of normal colonic mucosa.RESULTS:The expressions of KAI1 in HT29 and SW480cell lines were higher than those in LoVo and SW620.Theexpression of KAI1 gene was significantly higher in colorectalcarcinoma compared with normal colonic mucosa andlymphatic metastasis(χ~2=46.838,P<0.01).The expressionof KAI1 gene had no relationship with histological grade.The KAI1 expressions in Dukes A and B carcinoma werehigher at both mRNA and protein levels compared to DukesC carcinoma(χ~2=16.061,P<0.05).The expression of KAI1in colonic carcinoma specimens with lymphatic metastasiswas almost lost.The results of in situ hybridization werein concordance with immunohistochemistry.CONCLUSION:KAI1 is highly related to the metastasis ofcolonic carcinoma and may be a useful indicator of metastasisin colonic carcinoma.     

黏糖蛋白-2和组织蛋白酶-D与大肠癌恶性生物学行为的关系

目的了解黏糖蛋白-2(MUC-2)和组织蛋白酶-D(Cath-D)在大肠癌中的表达。方法免疫组织化学S-P法,检测60例大肠癌中MUC 2和Cath-D的表达情况。结果 MUC-2在大肠癌中的阳性率为70．00％,它的低表达与大肠癌的分期、分化程度和肠外转移呈正相关而与患者的性别、年龄、肿瘤的大体类型和淋巴转移与否等无关。Cath-D在大肠癌中的阳性率为66．67％,它的高表达与大肠癌的淋巴转移和肠外转移呈正相关而与患者的性别、年龄、肿瘤的大体类型、分期和分化程度等无关。结论 MUC-2和Cath-D的检测可成为大肠痛的恶性程度和肠外转移与否的重要参考指标。     

APPL1表达与结直肠癌发生发展的关系及意义

目的:研究APPL1蛋白在人结直肠癌组织中的表达情况与临床病理参数的关系.方法:收集35例新鲜结直肠癌及27例正常直肠黏膜组织,采用免疫组织化学SP法和RT-PCR法检测APPL1在结直肠癌组织及正常直肠黏膜组织中的表达.采用半定量积分分级对该蛋白的表达强弱进行评分.结果:免疫组织化学和RT-PCR结果显示,APPL1蛋白在结直肠癌组织以及正常直肠黏膜组织中普遍表达,但该蛋白和相应的mRNA在癌组织中的表达高于对照组(P<0.05).在35例结直肠癌组织中,APPL1表达与分化程度、淋巴结转移、TNM分期相关(P<0.05),与性别、年龄、肿瘤大小、组织学类型无明显相关(P>0.05).结论:APPL1蛋白在结直肠癌组织中的表达上调,且该蛋白表达与患者肿瘤的分化程度、淋巴结转移情况以及TNM分期有关.APPL1有可能成为结直肠癌治疗的一个新靶点.     

E-钙粘素、基质金属蛋白酶-2及其抑制剂与大肠癌浸润转移的关系

目的:探讨E-钙粘素(E-Cad)、基质金属蛋白酶-2(MMP-2)及其抑制剂(TIMP-2)表达与大肠癌浸润转移的关系。方法:采用S-P免疫组织化学染色技术,检测30例大肠腺癌,60例大肠癌组织E-Cad、MMP-2和TIMP-2的表达情况。结果:E-Cad的表达率在大肠腺瘤中为87.10%,显著高于大肠癌中的55.10%(P<0.05);其表达与大肠癌的大体类型有关,且随着大肠癌分化程度的降低而减少,与淋巴结转移呈负相关,E-Cad表达率越高患者的预后越好(P均<0.05)。MMP-2的表达率在大肠腺瘤中为26.67%,显著低于大肠癌中的86.67％(P<0.05);其表达与大肠癌的Dukes分期、分化程度、淋巴结转移和生存期均密切相关(P均<0.05)。TIMP-2的表达在大肠腺癌和大肠癌组织中没有显著性差异(P均>0.05),但其表达与大肠癌的Dukes分期、淋巴结转移、远隔脏器转移及生存期均有关(P均<0.05)。结论:E-Cad、MMP-2和TIMP-2的检测可以成为临床判断大肠癌的恶性程度、转移及预后的重要参考指标。     

Ezrin、FAK及E-cadherin在大肠癌组织中的表达及其临床意义

目的:探讨大肠癌组织中埃兹蛋白(Ezrin),黏着斑激酶(FAK)及上皮细胞钙黏蛋白(E-cadherin)的表达及其与肿瘤侵袭和转移的关系.方法:大肠癌组织标本50例,其中高分化腺癌13例,中低分化腺癌37例;无淋巴结转移30例,淋巴结转移20例.采用免疫组织化学方法检测50例大肠癌组织中Ezrin,FAK及E-cadherin的蛋白表达,并运用统计学方法分析Ezrin,FAK及E-cadherin的表达与大肠癌各种临床病理特征的关系及三者的相关性.结果:Ezrin在中低分化、伴有淋巴结转移及Dukes C D期大肠癌的阳性表达率显著高于高分化、无淋巴结转移及Dukes A B期大肠癌(83.78% vs 46.15%,P<0.01;95.00%vs 60.00%,P<0.01;95.00% vs 60.00%,P<0.01),而E-cadherin在以上组织中的表达正好相反(24.32% vs 69.23%,P<0.01;10.00% vs 53.33%,P<0.01;10.00% vs 53.33%,P<0.01),其均与患者性别及年龄大小均无关(P>0.05).FAK在Dukes C D期、伴有淋巴结转移大肠癌组织中的阳性表达显著高于Dukes A B、无淋巴转移组织(100.00% vs 63.33%,P<0.01;100.00% vs 63-33%,P<0.01),而与肿瘤的分化程度、患者性别及年龄大小均无关(P>0.05).经Spearman相关分析,Ezrin与FAK在大肠癌组织中的表达呈正相关(r=0.346,P<0.05),E-cadherin与Ezrin、FAK在大肠癌组织中的表达均呈明显负相关(r=-0.410,P<0.01;r=-0.406,P<0.01).结论:Ezrin,FAK及E-cadherin在大肠癌组织中的异常表达与肿瘤组织的浸润和转移密切相关,联合检测可以作为一组有效的大肠癌肿瘤标记和预后指标.     

YB-1和P53蛋白在结直肠肿瘤中的表达及意义

目的:探讨YB-1和P53蛋白在结直肠肿瘤不同发展阶段表达水平的差异及其与结直肠癌临床病理特征的关系.方法:采用免疫组织化学方法检测结直肠20例正常组织、30例低级别上皮内瘤变组织、30例高级别上皮内瘤变组织和50例癌组织中YB-1和P53蛋白的表达.结果:YB-1蛋白在结直肠正常组织(NCM)、低级别上皮内瘤变组织(LGIN)、高级别上皮内瘤变组织(HGIN)和癌组织(CRC)中的强阳性率分别为0%、76.7%、80.0%、80.0%,低级别、高级别上皮内瘤变组织和癌组织中的强阳性表达率均与正常组织比较差异有统计学意义(P<0.05),且在癌组织中其强阳性表达与淋巴结转移和TNM分期有关(P<0.05);P53蛋白在结直肠正常组织、低级别上皮内瘤变组织、高级别上皮内瘤变组织、癌组织中表达分别为0%、6.7%、40.0%、60.0%,高级别上皮内瘤变组织和癌组织中其表达率均与正常组织和低级别上皮内瘤变组织比较差异有统计学意义(P<0.05),其在癌组织中的表达与分化程度、淋巴结转移和TNM分期有关(P<0.05).在癌组织中YB-1和P53的表达呈正相关性(r=0.306,P<0.05).结论:YB-1和P53蛋白在结直肠癌的发生发展、转移中起重要作用,YB-1和P53蛋白的联合检测可能为判断结直肠癌恶性程度和转移提供重要参考.     

人结直肠癌中缺氧诱导因子-1α、血管内皮生长因子、生存素表达及意义

目的 探讨缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)、生存素表达在结直肠癌发生发展中的作用以及相互关系.方法 应用免疫组化法检测69例结直肠癌及20例正常肠黏膜中HIF-1α、VEGF、生存素的表达情况,并分析其与结直肠癌临床病理特征之间的关系.结果 HIF-1α、VEGF、生存素在结直肠癌中的表达率分别为56.52%、66.67%、46.38%,而在正常肠黏膜则表达不明显或不表达,两者差异有统计学意义(P＜0.01).HIF-1α、VEGF及生存素的表达与结直肠癌分化程度、肿瘤浸润深度、淋巴结转移和临床分期均密切相关(P＜0.05).HIF-1α蛋白的表达与VEGF及生存素蛋白的表达均呈显著正相关,VEGF与生存素的表达亦呈显著正相关.结论 HIF-1α通过上调VEGF及生存素表达促进结直肠癌的发生发展;VEGF与生存素间存在协同作用.     

KAI1 gene expression in colonic carcinoma and its clinical significances

AIM: To investigate KAI1 gene expression in the progression of human colonic carcinoma and its clinical significances. METHODS: KAI1 expression was detected by in situ hybridization and immunohistochemistry in the 4 established cell lines of colorectal carcinoma with different metastatic potentials, and in 80 specimens of colonic carcinoma, 21 colonic carcinoma specimens with lymphatic metastasis and 20 controls of normal colonic mucosa. RESULTS: The expressions of KAI1 in HT29 and SW480 cell lines were higher than those in LoVo and SW620. The expression of KAI1 gene was significantly higher in colorectal carcinoma compared with normal colonic mucosa and lymphatic metastasis (chi(2)=46.838, P<0.01). The expression of KAI1 gene had no relationship with histological grade. The KAI1 expressions in Dukes A and B carcinoma were higher at both mRNA and protein levels compared to Dukes C carcinoma (chi(2)=16.061, P<0.05). The expression of KAI1 in colonic carcinoma specimens with lymphatic metastasis was almost lost. The results of in situ hybridization were in concordance with immunohistochemistry. CONCLUSION: KAI1 is highly related to the metastasis of colonic carcinoma and may be a useful indicator of metastasis in colonic carcinoma.     

UQCRC1在结直肠癌及转移淋巴结中的表达

目的:分析结直肠癌(colorectal cancer,CRC)与正常黏膜间的蛋白质表达差异,筛选新的肿瘤标志物;并对兴趣蛋白进行验证,分析其与CRC的发生、发展及淋巴结转移的关系.方法:对6对新鲜的CRC与正常黏膜组织进行以二维差异凝胶电泳(2D differential gel electrophoresis,2D DIGE)及基质辅助激光解吸飞行时间质谱(matrix-assisted laserde sorption/ionization-time of flight masss pectrometry,MALDI-TOF-MS)分析.以免疫组织化学法验证兴趣蛋白泛醌细胞色素c还原酶核心蛋白1(ubiquinol cytochrome-creductase core protein 1,UQCRC1)在78例CRC与正常黏膜组织,和24个转移淋巴结中的表达.对染色的强弱评分为阴性:0,弱阳性:1,中阳性:2,强阳性:3.结果:2DDIGE分析显示在CRC中一个蛋白点丰度平均显著增高2.14倍(P<0.001).MALDI-TOF-MS分析证实该蛋白为UQCRC1.免疫组织化学法分析显示UQCRC1在CRC与正常黏膜组织中表达分别为2.28±0.95和1.81±0.88,有显著差异(P<0.001).UQCRC1表达的强弱与分化、分期及部位均无关(P>0.05).UQCRC1在转移淋巴结与配对的原发灶中表达分别为2.79±0.51和2.33±0.96,有显著差异(P<0.05).结论:UQCRC1在结直肠癌变和淋巴结转移过程中发挥一定的作用.     

Expression and significance of CD44s, CD44v6, and nm23 mRNA in human cancer

AIM: To investigate the relationship between the expression levels of nm23 mRNA, CD44s, and CD44v6,and oncogenesis, development and metastasis of human gastric adenocarcinoma, colorectal adenocarcinoma,intraductal carcinoma of breast, and lung cancer.METHODS: Using tissue microarray by immuhistochemical (IHC) staining and in situ hybri-dization (ISH), we examined the expression levels of nm23mRNA, CD44s, and CD44v6 in 62 specimens of human gastric adenocarcinoma and 62 specimens of colorectal adenocarcinoma; the expression of CD44s and CD44v6in 120 specimens of intraductal carcinoma of breast and 20 specimens of normal breast tissue; the expression of nm23 mRNA in 72 specimens of human lung cancer and 23 specimens of normal tissue adjacent to cancer.RESULTS: The expression of nm23 mRNA in the tissues of gastric and colorectal adenocarcinoma was not significantly different from that in the normal tissues adjacent to cancer (P＞0.05), and was not associated with the invasion of tumor and the pathology grade of adenocarcinoma (P＞0.05). However, the expression of nm23 mRNA was correlated negatively to the lymph node metastasis of gastric and colorectal adenocarcinoma (r = -0.49, P＜0.01; r = -4.93, P＜0.01). The expression of CD44s in the tissues of gastric and colorectal adenocarcinoma was significantly different from that in the normal tissues adjacent to cancer (P＜0.05;P＜0.01). CD44v6 was expressed in the tissues of gastric and colorectal adenocarcinoma only, the expression of CD44v6 was significantly associated with the lymph node metastasis, invasion and pathological grade of the tumor (r = 0.47, P＜0.01; r = 5.04, P＜0.01). CD44sand CD44v6 were expressed in intraductal carcinoma of breast, the expression of CD44s and CD44v6 was significantly associated with lymph node metastases and invasion (P＜0.01). However, neither of them was expressed in the normal breast tissue. In addition, the expression of CD44v6 was closely related to the degree of cell differentiation of intraductal carcinoma of breast (x2= 5.68, P＜0.05). The expressional level of nm23mRNA was closely related to the degree of cell differentiation (P＜0.05) and lymph node metastasis (P＜0.01), but the expression of nm23 gene was not related to sex, age, and type of histological classification (P＞0.05).CONCLUSION: Patients with overexpression of CD44s and CD44v6 and low expression of nm23 mRNA have a higher lymph node metastatic rate and invasion. In addition, overexpression of CD44v6 is closely related to the degree of cell differentiation. Detection of the three genes is able to provide a reliable index to evaluate the invasion and metastasis of tumor cells.