Nuclear β-catenin expression as a prognostic factor in advanced colorectal carcinoma

AIM： To investigate the changing pattern of β-catenin expression and its prognostic value in advanced colorectal cancer （CRC）. METHODS： Archival tumor samples were analyzed for β-catenin using immunohistochemistry （IHC） in 95 patients with advanced CRC. RESULTS： Membranous β-catenin expression was found in the normal colorectal epithelium. Almost 100% of CRC cases showed membranous and cytoplasmic expression, and 55 （58%） cases showed nuclear expression. In univariate （Kaplan-Meier） survival analysis, only the nuclear index （NI） was a significant predictor of disease free survival （DFS） （P = 0.023; n = 35）, with a NI above the median associated with longer DFS （34.2 too） than those with a NI below the median （15.5 too） （P = 0.045, ANOVA）. The other indices were not significant predictors of DFS, and none of the three tested indices （for membranous, cytoplasmic, or nuclear expression） predicted diseasespecific survival （DSS）. However, when dichotomized as positive or negative nuclear expression, the former was a significant predictor of more favorable DFS （P = 0.041） and DSS （P = 0.046）. CONCLUSION： Nuclear β-catenin expression provides additional information in predicting patient outcome in advanced CRC.     

Expression and clinical significance of high risk human papillomavirus and invasive gene in cervical carcinoma

Objective:To study the expression of E6 and E7 mRNA in high-risk human papillomavirus(HPV) HPV-18 and the relationship between the expression of invasive gene and cervical carcinoma.Methods:A total of 119 patients with cervical cancer,cervical erosion and cervical HPV infection who were diagnosed in our hospital were selected and randomly divided into two groups:cervical cancer group(n= 58) and non-cancerous group(n= 61).Another 60 patients with uterine leiomyoma were selected as normal control group.Detection of HPV18 E6,E7 mRNA expression and invasion,migration,proliferation inhibition genes,epithelial mesenchymal transition genes and proliferation related protein content.Results:The relative expression of E6 and E7 HPV-18 in cervical cancer group was significant higher than that in non-cancerous group and control group(mRNA)(P0.05).The content of TRAF6 and c-FLIP in invasive cervical cancer group was significantly higher than that in non-cancerous group and control group(P0.05).The mR NA content of CD44v6 and MMP-9 in cervical cancer group was significantly higher than that in non-cancerous group and control group(P0.05).The content of DEC-1,IKK16,MBP-1 in cervical cancer group was significant lower than that in non-cancerous group and control group(P0.05).The mR NA content of beta-catenin and Vimentin in cervical cancer group was significantly lower than that in non cancerous group and control group(P0.05).The proliferation related protein E2F1 of cervical cancer group was significantly lower than that of non-cancerous group and control group,Bmi-1 content was significantly higher than non-cancerous group and control group(P0.05).Conclusions:The expression of the detection of cervical cancer in high-risk human papilloma virus HPV-18 E6 and E7 mRNA,and the invasion,migration,proliferation inhibition gene,epithelial mesenchymal transition and proliferation related gene protein content,HPV expression rate of mR NA increased with the development of cervical cancer,the expression is also enhanced.The expression has a certain correlation between the level and development of cervical cancer.Through the above indicators,the development of cervical cancer monitoring and treatment to provide important clinical guidance.     

胃癌组织Maspin,uPA,MMP-7表达的意义

目的:观察胃癌及正常胃黏膜Maspin,uPA, MMP-7表达的意义．方法:应用免疫组化SP法检测胃管状腺癌30 例,胃印戒细胞癌30例,正常胃黏膜组织20例中Maspin,uPA,MMP-7的表达情况．结果:在胃管状腺癌中Maspin,uPA,MMP-7 阳性表达率分别为50％,70％和80％;胃印戒细胞癌中阳性表达率分别为46．7％,76．7％和 90％;正常胃黏膜组织中阳性表达率分别为 90％,35％和30％．Maspin的表达与浸润深度、淋巴结转移相关,而与肿块的大小和TNM分期无关．uPA和MMP-7的表达与浸润深度、淋巴结转移、TNM分期相关,而与肿块的大小无关．Maspin的表达与uPA和MMP-7的表达呈负相关(P=0．012,r=-0．322;P=0．008,r= -0．341);uPA的表达与MMP-7的表达呈正相关 (P=0．034,r=0．274)．结论:Maspin在胃癌中表达下调,uPA和 MMP-7在胃癌中过表达,他们在胃癌的浸润转移中起重要作用,可作为反应胃癌病理生物学行为的有效指标．     

血管生成素-2与基质金属蛋白酶-7在大肠癌中的表达及意义

目的:研究血管生成素-2(Ang-2)及基质金属蛋白酶-7(MMP-7)蛋白在人大肠癌组织中的表达及其与大肠癌临床病理特征的关系．方法:应用免疫组化法检测40例大肠癌及其癌旁正常组织中Ang-2及MMP-7蛋白表达水平．结果:大肠癌组织中Ang-2蛋白表达阳性率为77．5％(31/40),明显高于癌旁正常组织 40％(16/40)(P=0．001);Ang-2表达水平与患者性别、年龄及癌组织的部位、大小、分化程度、淋巴结转移无关(P>0．05);与浸润深度, 远处转移及Dukes’分期相关(分别为P=0．007, P=0．023,P=0．008);大肠癌组织RMMP-7蛋白表达阳性率为85％(34/40),明显高于癌旁正常组织35％(14/40)(P=0．000)．MMP-7蛋白表达阳性率与Ang-2的表达相关(P=0．016)．结论:Ang-2蛋白可能通过促进MMP-7的表达从而促进了大肠癌的生长及转移．     

The metastasis-associated gene S100A4 is a novel target of beta-catenin/T-cell factor signaling in colon cancer

BACKGROUND & AIMS: Activation of the Wnt/beta-catenin pathway is frequently observed in colorectal cancers. Our aim was to elucidate the impact of gain-of-function beta-catenin on the metastasis-associated gene S100A4 in human colon cancer cell lines and tumors. METHODS: We analyzed cell lines heterozygous for gain-of-function and wild-type beta-catenin, and variants homozygous for gain- or loss-of-function mutation in beta-catenin, for S100A4 expression, cell motility, and in vivo metastasis. beta-catenin-mediated S100A4 promoter activation was tested by reporter assays. For human colon carcinomas, S100A4 expression, beta-catenin genotype, and metachronous metastasis were correlated. RESULTS: We identified S100A4 as the most regulated gene by gain-of-function beta-catenin using a 10K microarray. Cell lines with gain-of-function beta-catenin expressed up to 60-fold elevated S100A4 levels, displayed strongly increased migration and invasion in vitro, and induced metastasis in mice. S100A4 small interfering RNA, beta-catenin small interfering RNA, or dominant negative T-cell factor (TCF) knocked down S100A4 and blocked biological effects. S100A4 complementary DNA transfection increased migration and invasion. We identified a TCF binding site within the S100A4 promoter and demonstrated the direct binding of heterodimeric beta-catenin/TCF complexes. Reporter assays confirmed the beta-catenin-induced S100A4 promoter activity. Furthermore, S100A4 mRNA expression was increased in primary colon cancers, which later developed distant metastases, compared to non-metastasizing tumors. Colon tumors heterozygous for gain-of-function beta-catenin showed concomitant nuclear beta-catenin localization, high S100A4 expression, and metastases. CONCLUSIONS: S100A4 is a direct beta-catenin/TCF target, induces migration and invasion in vitro and metastasis in vivo, and has value for prognosis of metastasis formation in colon cancer patients.     

结肠癌组织中MMP-7、MMP-9的表达及意义

目的观察结肠癌组织中基质金属蛋白酶（MMP）-7、MMP-9的表达，探讨其在结肠癌浸润、转移中的作用。方法采用免疫组化SP法检测50例结肠癌及其切缘组织中MMP-7、MMP-9的表达。结果肿瘤组织中MMP-7、MMP-9阳性表达率分别为68．00％和82．00％，标本切缘组织中分别为0和24．00％，两者相比，P均〈0．05，MMP-7、MMP-9在有淋巴结转移者中的阳性表达率为86．96％和91．30％，显著高于无淋巴结转移者的51．85％和74．07％，P均〈0．05；MMP-7、MMP-9在结肠癌组织中的表达呈正相关，r=0．84，P〈0．05。结论MMP-7、MMP-9高表达可能与结肠癌的浸润、转移有关。     

Gene-expression profiling predicts recurrence in Dukes' C colorectal cancer

BACKGROUND & AIMS: Although approximately 50% of Dukes' C colorectal cancer patients are surgically cured, it is currently not possible to distinguish these patients from those at high risk of recurrence. The recent advent of routine adjuvant chemotherapy for these patients has greatly complicated the identification of new markers predicting the response to surgery, which is now reliant on archived materials. Microarray analysis allows fine tumor classification but cannot be used with paraffin-embedded archival samples. METHODS: We used microarray analysis of a unique set of fresh-frozen tumor samples from Dukes' C patients who had surgery as the only form of treatment to identify molecular signatures that characterize tumors from patients with good and bad prognosis. RESULTS: Unsupervised hierarchical clustering and a K-nearest neighbors-based classifier identified groups of patients with significantly different survival (P = .019 and P = .0001). Expression profiling outperformed previously reported genetic markers of prognosis such as TP53 and K-RAS mutational status and allelic imbalance in chromosome 18q, which were of limited prognostic power in this study. Functional categories significantly enriched in gene-expression differences included protein transport and folding. The prognostic potential of the RAS homologue RHOA, one of the most differentially expressed genes, was further investigated using immunohistochemistry and a tissue microarray containing 137 independent Dukes' C tumor samples. Reduced RHOA expression was associated with significantly shorter survival (P = .01). CONCLUSIONS: This study shows that gene-expression profiling of surgical tumor samples can predict recurrence in Dukes' C patients. Therefore, this approach could be used to guide decisions concerning the clinical management of these patients.     

Up-regulation of α-catenin is associated with increased lymph node involvement in colorectal cancer

AIM： To investigate the changing pattern of α-catenin expression and its relationship to clinical and pathological features of colorectal cancer （CRC） patients. METHODS： Archival tumor samples were analyzed using immunohistochemistry （IHC） for α-catenin in 91 patients with advanced CRC. RESULTS： The values of α-catenin membrane index （MI） and cytoplasmic index （CI） were significantly related to the depth of tumor invasion （P = 0.027, P = 0.020, respectively）, high indices being associated with increased depth of the primary tumor invasion （T3 and T4）. Similarly, patients with high α-catenin expression had a significantly increased risk of lymph node metastasis （32/39 vs 37/52 for MI and 37/45 vs 32/46 for CI） （P = 0.001, P = 0.0001, respectively, for LNN status）. An altered expression （i.e., cytoplasmic pattern） was also related （P = 0.047） to the response to chemotherapy; patients with low CI were more responsive （CR： 7/46） than patients with high CI values （CR： 0/45）. There was a marginal effect on survival in patients time with metastases （SWM） （P = 0.087）; patients with low CI showing slightly longer SWM, but no such effect on disease free survival （DFS） or disease specific survival （DSS）. As to co-expression with another member of the adhesion complex （β-catenin）, high α-catenin/β-catenin MI index was of marginal significance in predicting longer DSS （P = 0.063, log-rank）. CONCLUSION： The results implicate that high α-catenin expression is intimately involved in the key regulatory mechanisms leading to invasive phenotype, lymph node metastases, and progressive disease in CRC.     

Expressions of inducible nitric oxide synthase and matrix metalloproteinase-9 and their effects on angiogenesis and progression of hepatocellular carcinoma

AIM: To determine the expressions of inducible nitric oxide synthase (iNOS) and matrix metalloproteinase-9 (MMP-9) in hepatocellular carcinoma (HCC) and to investigate the relationship between iNOS and MMP-9 expression and their effects on angiogenesis and progression of HCC. METHODS: In this study, we examined iNOS, MMP-9, and CD34 expression in specimens surgically removed from 32 HCC patients and 7 normal liver tissues by immunohistochemical staining. Meanwhile, microvessel density (MVD) was determined as a marker of angiogenesis by counting CD34-positive cells. RESULTS: The positive rates of iNOS and MMP-9 expression were 71.88% (23/32) and 78.13% (25/32) in HCC. MMP-9 expression was significantly correlated with tumor size, capsule status, TNM stage, and risk of HCC recurrence (P= 0.032, P= 0.033, P= 0.007, and P= 0.001, respectively). There was also a significant relationship between iNOS expression and capsule status and risk of HCC recurrence (P= 0.049 and P= 0.004, respectively), but no correlation between iNOS expression and tumor size and TNM stage. There was a positive association between MVD and TNM stage and risk of HCC recurrence (P= 0.037 and P= 0.000, respectively). The count of MVD was significantly different in different iNOS and MMP-9 immunoreactivity groups (F= 17.713 and 17.097, P= 0.000 and P= 0.000, respectively). The examination of Spearman's rank correlation coefficient showed that there was a significant positive correlation between MVD and iNOS, MMP-9 immunoreactivity (r= 0.754 and 0.751, P= 0.000 and A=0.000, respectively). There was also a significant association between MMP-9 and iNOS expression in HCC (P= 0.010). CONCLUSION: Nitric oxide (NO) produced by iNOS could modulate MMP-9 production and therefore contribute to tumor cell angiogenesis and invasion and metastasis in HCC. The strong expression of iNOS and MMP-9 in HCC may be helpful in evaluating the recurrence of HCC, predicting poor prognosis. For patients with strong expression of MMP-9 and iNOS, the optimal treatment scheme needs to be selected.     

Prognostic significance of computed tomography-detected extramural vascular invasion in colon cancer

AIM: To compare disease-free survival(DFS) between extramural vascular invasion(EMVI)-positive and-negative colon cancer patients evaluated by computed tomography(CT).METHODS: Colon cancer patients(n = 194) undergoing curative surgery between January 2009 and December 2013 were included. Each patient's demographics, cancer characteristics, EMVI status, pathological status and survival outcomes were recorded. All included patients had been routinely monitored until December 2015. EMVI was defined as tumor tissue within adjacent vessels beyond the colon wall as seen on enhanced CT. Disease recurrence was defined as metachronous metastases, local recurrence, or death due to colon cancer. Kaplan-Meier analyses were used to compare DFS between the EMVI-positive and-negative groups. Cox's proportional hazards models were used to measure the impact of confounding variables on survival rates.RESULTS: EMVI was observed on CT(ct EMVI) in 60 patients(30.9%, 60/194). One year after surgery, there was no statistically significant difference regarding the rates of progressive events between EMVI-positive and-negative patients [11.7%(7/60) and 6.7%(9/134), respectively; P = 0.266]. At the study endpoint, the EMVI-positive patients had significantly more progressive events than the EMVI-negative patients [43.3%(26/60) and 14.9%(20/134), respectively; oddsratio = 4.4, P 0.001]. Based on the Kaplan-Meier method, the cumulative 1-year DFS rates were 86.7%(95%CI: 82.3-91.1) and 92.4%(95%CI: 90.1-94.7) for EMVI-positive and EMVI-negative patients, respectively. The cumulative 3-year DFS rates were 49.5%(95%CI: 42.1-56.9) and 85.8%(95%CI: 82.6-89.0), respectively. Cox proportional hazards regression analysis revealed that ctE MVI was an independent predictor of DFS with a hazard ratio of 2.15(95%CI: 1.12-4.14, P = 0.023). CONCLUSION: ctE MVI may be helpful when evaluating disease progression in colon cancer patients.     

β-catenin up-regulates the expression of cyclinD1, c-myc and MMP-7 in human pancreatic cancer： Relationships with carcinogenesis and metastasis

AIM: To investigate whether abnormal expression of β-catenin in conjunction with overexpression of cyclin D1, c-myc and matrix metalloproteinase-7 (MMP-7) correlated with the carcinogenesis, metastasis and prognosis of pancreatic cancer, and to analyze the relationship of β-catenin expression with cyclinD1, c-myc and MMP-7 expression.METHODS: Using immunohistochemistry, we examined the expression of β-catenin, cyclinD1, c-myc and MMP-7 in 47 pancreatic adenocarcinoma tissues, 12 pancreatic intraepithelial neoplasia (PanIN) and 10 normal pancreases,respectively. Proliferation cell nuclear antigen was also tested as the index of proliferative activity of pancreatic cancer cells.RESULTS: In 10 cases of normal pancreatic tissues,epithelial cells showed equally strong membranous expression of βcatenin protein at the cell-cell boundaries,but the expression of cyclin D1, c-myc and MMP-7 was negative. The expression of βcatenin, cyclinD1, c-myc and MMP-7 in PanIN and pancreatic adenocarcinoma tissues had no significant difference [6/12 and 32/47 (68.1%),6/12 and 35/47 (74.5%), 5/12 and 33/47 (70.2%), 7/12 and 30/47 (63.8%), respectively]. The abnormal expression of β-catenin was significantly correlated to metastasis and one-year survival rate of pancreatic cancer, but had no relation with size, differentiation and cell proliferation.The expression of cyclinD1 was correlated with cell proliferation and extent of differentiation, but not with size,metastasis and one-year survival rate of the pancreatic ancer. The expression of c-myc was not correlated with ize, extent of differentiation, metastasis and 1-year urvival rate, but closely with cell proliferation of pancreatic ancer. The overexpression of MMP-7 was significantly ssociated with metastasis and 1-year survival rate of ancreatic cancer, but not with size, extent of differentiation and cell proliferation. There was a highly significant positive association between abnormal expression of β-catenin and overexpression of cyclinD1, c-myc and MMP-7 not only in PanIN (r = 1.000, 0.845, 0.845), but also in pancreatic cancer (r = 0.437, 0.452, 0.435).CONCLUSION: The abnormal expression of β-catenin plays a key role in the carcinogenesis and progression of human pancreatic carcinoma by up-regulating the expression of cyclinD1, c-myc and MMP-7, resulting in the degradation of extracellular matrix and uncontrolled cell proliferation and differentiation, β-catenin abnormal expression and MMP-7 overexpression may be considered as two useful markers for determining metastasis and prognosis of human pancreatic cancer.     

异常隐窝病灶与大肠肿瘤关系及发生途径的初步研究

目的 实体镜下连续观察Wistar鼠大肠癌模型中异常隐窝病灶(ACF)的发生情况,探讨ACF与大肠肿瘤的相关性及其发生途径.方法 60只Wismr鼠给予二甲肼皮下注射,每周1次,连续18周,分组处死.将美蓝染色后的大肠组织在实体镜下观察.结果 发现2种不同镜下表现的ACF,即cACF和dACF.dACF与大肠肿瘤形成早期有相似的镜下形态及病理特点,而cACF则无类似特征.cACF与dACF在β-catenin中的表达异常率分别为4.8%和100.0%(P=0.000),在MMP-7中的阳性表达率分别为7.9%和81.8%(P=0.000),dACF与肿瘤在β-catenin的表达异常率以及在MMP-7的阳性表达率上的差异均无统计学意义(P>0.05).结论 cACF与肿瘤发生无明显相关,dACF与肿瘤发生关系密切,其发生遵循Wnt途径.     

大肠癌血管内皮生长因子的表达及其意义

目的：探讨血管内皮生长因子（VEGF）mRNA在大肠癌和癌周组织中的表达及其与临床特征之间的关系,为在分子水平干预肿瘤血管形成,预防大肠癌复发和转移打下基础。方法：采用逆转录－聚合酶链反应（RT－PCR）检测方法,对68例大肠癌手术标本癌和癌周组织中VEGF mRNA的表达水平进行了相对定量研究。结果：肿瘤组织中VEGF mRNA的表达率为67．6％（46／68）,癌周组织表达率为32．4％（22／68）;肿瘤组织中VEGF mRNA表达水平在浆膜浸润组,淋巴结转移,远处转移和Dukes D期组分别显著高于未侵及浆膜组,无淋巴结转移组,无远处转移和Dukes A,B,C期组;肿瘤组织中VEGF mRNA表达水平在肿瘤直径大的大肠癌组（＞5cm)与大小肠癌组（≤5cm)以及不同组织学分组组之间相比差异无显著性。结论：VEGF在大肠癌浸润和转移过程中发挥重要作用,肿瘤血管形成与大肠癌浸润和转移关系密切。     

Prognostic significance of BMP and activin membrane-bound inhibitor in colorectal cancer

AIM： To investigate the clinical significance of BMP and activin membrane-bound inhibitor （BAMBI） which is a pseudoreceptor of transforming growth factorbeta （TGF-β） type 1 receptors and acts as a negative regulator of TGF-β signaling and expression aberrantly elevated in colorectal cancers （CRCs）. We studied BAMBI expression in CRCs. METHODS： We studied BAMBI expression in 183 surgically resected CRCs by immunochemical and immunoblotting analyses using a generated monoclonal anti-BAMBI antibody. Commercially available anti-β- catenin and anti-p53 antibodies were also applied for immunochemical analyses as a comparison control.RESULTS： Immunohistochemical analysis revealed that BAMBI expression was observed in 148 （80.8%）, and strong BAMBI expression was observed in 46% of the CRCs. Strong BAMBI expression was positively correlated with histological type, depth of invasion, lymph node metastases, and tumor node metastasis （TNM） stage （P 〈 0.05）. Clear associations were found between BAMBI and β-catenin （P = 0.035） and p53 （P =0.049） expression. In curatively resected CRC, 5-year recurrence-free survival was 51.9% （P = 0.037） for strong BAMBI expression compared to 79.8% for weak BAMBI expression. In the Cox＇s multivariate analysis, lymph node metastases （relative risk 6.685; P 〈 0.001） and depth of invasion （RR 14.0; P = 0.013） were significant indicators for recurrence, and strong BAMBI expression （RR 2.26; P = 0.057） tended to be significant. CONCLUSION： BAMBI was linked to a potentially aggressive tumor phenotype and predicted tumor recurrence and cancer-related death in CRC. BAMBI expression might be applicable in the routine clinical setting of CRC.