High Pin1 expression is associated with tumor progression in colorectal cancer

BACKGROUND AND OBJECTIVES: Peptidyl prolyl cis-trans isomerase (Pin1) isomerizes only phosphorylated serine or threonine residues preceding proline in certain proteins and affects the protein function. Pin1 interacts with many signaling pathways, including Wnt signaling pathway that is crucial for colorectal tumorigenesis. Pin1 promotes cyclin D1 over-expression directly or through the stabilization of beta-catenin. Pin1 is over-expressed in some cancers such as prostate and breast cancers. This study aimed to determine whether Pin1 plays a role in colorectal tumorigenesis through the upregulation of beta-catenin and cyclin D1. METHODS: Immunohistochemical analyses were performed on 105 colorectal cancer tissue samples using anti-Pin1, anti-beta-catenin, and anti-cyclin D1 antibodies. We examined the relationships between Pin1 expression and clinicopathological factors, prognosis, and beta-catenin/cyclin D1 expression. RESULTS: High Pin1 expression was observed in 40 cases (38%) and positively correlated with histological type (P=0.0240), depth of invasion (P=0.0051), and staging (P=0.0027) of colorectal tumors. High Pin1 expression was also correlated with the over-expressions of both beta-catenin (P=0.0225) and cyclin D1 (P=0.0137). CONCLUSIONS: These results suggest that Pin1 plays an important role in colorectal tumorigenesis, presumably by increasing beta-catenin and cyclin D1 expressions.     

Significance of E-cadherin/beta-catenin complex and cyclin D1 in breast cancer

E-cadherin and beta-catenin are important epithelial adhesion molecules in normal epithelium. Loss of E-cadherin - beta-catenin adhesion is an important step in the progression of many epithelial malignancies. beta-catenin plays also a role in intracellular signaling and can function as an oncogene when binds to the T-cell factor 4 (Tcf4)-binding site in the promotor region of cyclin D1 and transactivates genes after translocation to the nucleus. We evaluated the immunohistochemical expression pattern of E-cadherin, beta-catenin in relationship with cyclin D1 overexpression, tumor stage, clinicopathologic parameters and patient survival in 128 mammary infiltrating duct carcinomas. The expression of E-cadherin/beta-catenin complex and beta-catenin/cyclin D1 double staining with confocal scanning laser microscope was evaluated. There were aberrant expressions in 78% of E-cadherin, 79% of beta-catenin, and 66% of cyclin D1 in breast cancer. There was correlation of aberrant expression of E-cadherin or beta-catenin with lymph node metastasis, survival rate, and survival length. However, there was no correlation of cyclin D1 overexpression with aberrant expression of E-cadherin or beta-catenin. No death was found in normal expression of beta-catenin, however lowest survival (50%) was found in nuclear beta-catenin expression. There was correlation of overexpression of cyclin D1 with survival rate and survival length. The highest survival rate and survival length were found in membranous normal beta-catenin expression group, however significant decrement of survival length was found in the groups of aberrant expression one or both of E-cadherin or/and beta-catenin. These results suggest that aberrant expression of E-cadherin, beta-catenin, and cyclin D1 may be involved in tumor metastasis, and analysis of the degree or the pattern of E-cadherin, beta-catenin, cyclin D1, and E-cadherin/beta-catenin complex may be good prognostic markers of mammary infiltrating duct carcinoma.     

Pin1、β-连接素和细胞周期素D1在老年肺癌组织中的表达及临床意义

Objective:To investigate the expressions and correlations of Pin1,β-catenin and cyclin D1 in elderly lung carcinomas.Methods:The expressions of Pin1,β-catenin and cyclin D1 were examined in the specimens of 92 elderly lung carcinomas and 10 normal lung tissues by immunohistochemistry and explored the relationship between the expression levels and clinicopathological factors.Results:(1) The overexpression of Pin1 and cyclin D1 in lung carcinomas was 46 (50%)cases and 60 (65.22%) cases respectively and 56 (60.82%) cases showed positive immunoreactivity for β-catenin in the nuclear and (or) cytoplasmic fraction in tumor tissues,In normal tissue,the expressions of Pin1 and cyclin D1 were negative,the expression of β-catenin was lied in cell membrane.(2) In lung carcinomas the expressions of Pin1,β-catenin and cyclin D1 correlated with tumor differentiation (P＜0.05).The pesitive expression rate and intensity of Pin1 correlated with tumor stage (P=0.032) and lymph node positive disease (P=0.041).The expression of β-catenin correlated with lymph node positive disease (P=0.012).(3) High expression levels of Pin1 correlated with aberrant β-catenin expression (P=0.000) but did not show a correlation with cyclin D1 (P=0.157).Conclusion:In elderly lung carcinomas,the positive expression of Pin1 causes abnormal accumulation of β-catenin and actives its target gene,however,this target gene was not cyclin DI.The detection of Pin1 expression had some clinical significance in estimating prognosis of elderly patient with lung carcinomas.     

乳腺癌组织中E-cadherin、β-catenin及cyclin D1表达的相关性研究

背景与目的上皮性钙粘素(E-cadherin)通过连接素(catenins)与细胞骨架相连介导细胞同质粘附反应,β-catenin除与E-cadherin结合介导细胞粘附反应外,还作为Wnt信号转导通路的重要成分与肿瘤发生密切相关。本研究通过检测乳腺癌组织中E-cadherin、β-catenin及cyclinD1的表达,探讨E-cadherin、β-catenin在乳腺癌发生、发展中的意义。方法采用免疫组织化学SP法检测60例乳腺癌组织中E-cadherin、β-catenin、cyclinD1的表达。结果乳腺癌组织中有29例(48.3%)E-cadherin、18例(30.0%)β-catenin正常表达,28例(46.7%)cyclinD1过度表达。E-cadherin正常表达病例中,31.0%(9/29)的病例呈现cyclinD1过度表达,而E-cadherin异常表达病例中,61.3%(19/31)的病例呈现cyclinD1过度表达,E-cadherin异常表达与cyclinD1的过度表达有显著的正相关性(rs=0.303,P<0.05)。有42例癌组织表现出β-catenin的异常表达,其中57.1%(24/42)的病例出现cyclinD1的过度表达,而β-catenin正常膜表达病例中,22.2%(4/18)的病例呈现cyclinD1的过度表达。β-catenin的异常表达与cyclinD1的过度表达有显著的正相关性(rs=0.321,P<0.05)。结论E-cadherin和β-catenin的异常表达可能通过促使或激活cyclinD1的过度表达导致乳腺癌的发生和发展。     

Overexpression of Pin1 in non-small cell lung cancer (NSCLC) and its correlation with lymph node metastases

BACKGROUND: Pin1 isomerizes the bonds of molecules important for numerous oncogenic and cell-signaling pathways, including Bcl-2, p53, c-Jun, beta-catenin, NF-kappaB, cyclin D1, c-Myc and Raf-1. This can cause a change in conformation leading to alterations in catalytic activity, protein-protein interactions, subcellular localization and protein stability. These alterations have been shown to be associated with cell transformation and cancer progression. Pin1 is overexpressed in several different human cancers. This is the first report of Pin1 overexpression in clinical samples of non-small cell lung cancer (NSCLC). METHODS: Protein expression levels of Pin1 in tumor and normal lung specimens were analyzed for expression of Pin1, cyclin D1, p53 and MDM2 using immunohistochemistry and compared to several clinicopathological characteristics. The mRNA expression of Pin1 was also analyzed using quantitative real-time RT-PCR and compared to clinicopathological characteristics. RESULTS: Pin1 protein was shown to be overexpressed in NSCLC tumor samples, and correlated with lymph node positive disease and tumor stage. High expression of MDM2 also correlated with lymph node positive disease and with poorly differentiated tumors. High expression of MDM2 also correlated with lymph node positive disease and with poorly differentiated tumors. High expression levels of Pin1 correlated with high levels of p53 or MDM2 protein, but did not show a correlation with cyclin D1. However, high levels of MDM2 correlated with cyclin D1 overexpression. Pin1 mRNA was expressed significantly more often in the tumors of smokers than of non-smokers. The relationship between the expression of protein and mRNA of Pin1 has obviously showed that protein expression isn't significantly associated with mRNA expression. CONCLUSIONS: Pin1 is overexpressed in many different cancers, including NSCLC, and may possibly be used as a tumor marker or as a target for cancer therapy.     

宫颈癌细胞株和宫颈上皮组织中Pin1和Cyclin D1的表达及临床意义

背景与目的：肽基脯氨酰顺反异构酶Pin1在人类多种肿瘤中过表达,Pin1过表达可增强Cyclin D1表达,在肿瘤发生发展中起重要作用。本研究拟探讨宫颈癌细胞株及宫颈上皮组织中Pin1、Cyclin D1的表达及其临床意义。方法：应用RT-PCR、Western blot法检测HeLa、SiHa、C33a、Caski细胞中Pin1和Cyclin D1基因蛋白表达,免疫组化法检测10例正常宫颈、21例宫颈上皮内瘤样病变、57例浸润癌组织中Pin1和Cyclin D1的表达状况,评价其临床意义。结果：HeLa、SiHa、C33a、Caski细胞存在不同程度Pin1 mRNA及蛋白过表达（P〈0．05）。正常宫颈、宫颈上皮内瘤样病变、浸润癌组织中Pin1蛋白表达逐渐增强,分别为0％、47．62％、64．91％（P〈0．05）。Pin1蛋白过表达与宫颈癌临床分期、病理分级、淋巴结转移无关（P〉0．05）;但宫颈腺癌组织中Pin1表达明显高于鳞癌（P〈0．05）。Pin1过表达与Cyclin D1表达呈显著正相关（P〈0．05）。结论：Pin1在宫颈癌细胞株及宫颈癌组织中存在mRNA及蛋白水平上的过表达。Pin1过表达与Cyclin D1表达密切相关．Pin1和Cyclin D1异常表达可能参与宫颈癌的发生发展。     

Beta-catenin and cyclin D1 expression in human hepatocellular carcinoma

To understand the nature and roles of mutated beta-catenin in human hepatocellular carcinomas (HCCs), 57 cases of surgically resected HCCs were studied. DNAs extracted from each tumor were examined for somatic mutations of exon 3, and the protein expressions of beta-catenin, cyclin D1, and Ki-67 were observed by immunohistochemical staining. beta-catenin mutations in exon 3 were detected in 10 (17.5%) out of 57 HCCs, including nine missense mutations and one deletion mutation. All of the cases with gene alterations had the anti-HCV antibody, and tested negative for the HBs antigen in the sera. All of the mutations occurred at the serine/threonine phosphorylation sites of glycogen synthase kinase-3beta (GSK-3beta) or their neighboring residues. Significant correlation with intracellular expression (p=0.00055) was shown in the HCCs harboring beta-catenin mutations. The intracellular accumulation of beta-catenin showed significant correlation with the cyclin D1 expression (p=0.00858), and with a higher proliferation index (p=0.00072). In addition, the beta-catenin mutations showed significant association with the cyclin D1 expression (p=0.0424). These results suggest that accumulated beta-catenin proteins may bind to the lymphocyte enhancer binding factor-1 (LEF-1), form the beta-catenin/LEF-1 complex, and stimulate such promoters regulating the cell cycle as the cyclin D1 gene. This is the first report to demonstrate a significant correlation between beta-catenin and the cyclin D1 expression in human HCCs.     

非小细胞肺癌cyclin D1基因表达与预后关系的初步探讨

目的 探讨非小细胞肺癌ｃｙｃｌｉｎ Ｄ１基因表达与预后的关系。方法 采用免疫组化Ｅｎｖｉｓｉｏｎ二步法对４６例非小细胞肺癌手术标本进行ｃｙｃｌｉｎ Ｄ１检测,并结合临床随方资料进行分析。结果 ２５例ｃｌｃｉｎ Ｄ１表达阳性,表达率为５４．３％;其中细胞核阳性表达６例,细胞质阳性表达１９例。临床随访资料显示,在ｃｙｃｌｉｎ Ｄ１基因表达阳性的２５例患者中有１０例术后发生远外转移,而ｃｙｃｌｉｎ Ｄ１     

β-连环蛋白在食管癌组织中的表达及意义

背景与目的:许多研究已经证实β-连环蛋白(beta-catenin)在多种人类肿瘤中有异常表达。为确定β-连环蛋白是否参与食管癌的发生发展,本实验检测β-连环蛋白在正常食管上皮及食管癌组织中的表达状态,研究其表达、定位与肿瘤分化程度及淋巴结转移的关系。方法:采用免疫组化SP法,检测52例食管癌及22例正常食管组织切片中β-连环蛋白的表达。结果:22例正常食管组织中,β-连环蛋白表达均显著位于细胞膜,而胞浆表达很少见;52例食管癌组织中,β-连环蛋白均呈现不同程度的胞浆表达,同时伴随细胞膜表达量减少甚至缺失,其中14例在细胞核内还可检出β-连环蛋白。另外,在有淋巴结转移的食管癌病例中,β-连环蛋白胞浆表达的阳性率明显高于无淋巴结转移的食管癌病例(P<0.05)。结论:β-连环蛋白在食管癌组织中表达异常较为普遍,主要表现在蛋白从细胞膜到细胞浆、到细胞核的转位,而且胞浆累积明显;这种改变与淋巴结转移率显著相关。     

The Positive Nuclear Staining Observed with Monoclonal Antibody against PRAD1/Cyclin D1 Correlates with mRNA Expression in Mantle Cell Lymphoma

Recently, we produced a monoclonal antibody, 5D4, against the PRAD1/cyclin D1 product and suggested positive nuclear staining to be associated with mantle cell lymphoma (MCL). Now we have further characterized the specificity of this antibody and studied the relation of immunohistochemical detection to PRAD1/cyclin D1 mRNA expression and DNA rearrangement. Immunofluorescence and immunoblotting studies demonstrated the 5D4 antibody to be crossreactive with cyclin D2, but not cyclin D3. On immunostaining, 15 of 19 MCL cases (79%) presented the nuclear staining pattern and PRAD1/cyclin D1 mRNA expression was detected by Northern blot analysis in 12 of 15 MCL cases studied (80%); all cases with the mRNA expression showed the nuclear staining pattern. Southern blot analysis with 11q13 BCL-1 probes detected DNA rearrangements in 8 of 19 MCL cases (42%), all 8 exhibiting PRAD1/cyclin D1 mRNA expression. In 21 lymphoma cases of types other than MCL, neither the mRNA expression nor the nuclear staining were observed, although cytoplasmic staining was often apparent. These results indicated that positive nuclear staining of lymphoma cells by 5D4 antibody reflects PRAD1/cyclin D1 mRNA expression, and showed that this monoclonal antibody has diagnostic value for differentiating MCL from other types of lymphomas.     

cyclin D1在喉癌组织中的表达研究

目的　探讨cyclinD 1在喉癌组织中的表达及其临床意义。方法　采用免疫组织化学S P法检测 41例喉癌组织中cyclinD 1的表达 ,并以 12例癌旁正常组织作对照 ,进行比较分析。 结果　 41例喉癌组织中cyclinD 1阳性表达率为 5 1.2 % ,12例癌旁正常组织阳性表达率为 16.7% ,两者有显著性差异 (P <0 .0 5 )。cyclinD1的阳性表达率与喉癌患者的性别、年龄、病程、吸烟、饮酒、肿瘤部位和TNM分期无关 (P >0 .0 5 ) ,但与淋巴结有无转移以及喉癌组织的病理分级有显著相关性 (P <0 .0 1)。结论　cy clinD1在喉癌的发生发展过程中起着一定的作用 ,可作为判断喉癌恶性程度和预后的 1个重要参考指标。     

Cyclin D1 overexpression related to retinoblastoma protein expression as a prognostic marker in human oesophageal squamous cell carcinoma.

The relationship between aberrant expression of cyclin D1 and retinoblastoma (RB) protein and clinicopathological factors was investigated in 80 patients with oesophageal SCC using immunohistochemical analyses. Heterogeneous staining of cancer cell nuclei with antibody to cyclin D1 was found in 31.3% of patients (25 out of 80 patients). Nuclear staining of cancer cells with anti-RB antibody was homogeneous in 10.0% (8 out of 80 patients) and heterogeneous in 58.8% (47 out of 80 patients). Among cases with homogeneous staining for RB protein, 75% (six out of eight patients) exhibited simultaneous positivity for cyclin D1 (P < 0.05). No significant relationship was found between cyclin D1 or RB protein expression and various clinicopathological parameters. The prognosis of patients with cyclin D1-positive tumours was significantly poorer than that of the other patients (P < 0.01). In addition, when patients with cyclin D1-positive and -negative tumours were stratified according to presence or absence of lymph node metastasis and RB status, the cumulative survival rates in the cyclin D1-positive groups were significantly lower for patients without lymph node metastasis (P < 0.01) and for patients whose tumours were positive for RB (P< 0.0001). These findings suggest the possibility that cyclin D1 positivity is a useful prognostic marker related to lymph node metastasis and RB protein expression in human oesophageal SCC, in addition to clinicopathological factors.     

Expression and subcellular localization of cyclin D1 protein in epithelial ovarian tumour cells

The expression of cyclin D1 protein in tumour sections from 81 patients with epithelial ovarian cancer was analysed using immunohistochemistry. The tumours that overexpressed cyclin D1 in more than 10% of neoplastic cells were considered positive. Thus overexpression of cyclin D1 was observed in 72/81 (89%) of the cases examined. Protein was detected in both the nucleus and the cytoplasm in 24/81 (30%) and localized exclusively in the cytoplasm in 48/81 (59%) of the tumours. Cyclin D1 was overexpressed in both borderline and invasive tumours. There was no association between protein overexpression and tumour stage and differentiation. Furthermore, no correlation between cyclin D1 expression and clinical outcome was observed. However, in tumours overexpressing cyclin D1 (n = 72), the proportion displaying exclusively cytoplasmic localization of protein was higher in those with serous compared with non-serous histology (P = 0.004, odds ratio 4.8, 95% confidence interval 1.4-19.1). Western analysis using a monoclonal antibody to cyclin D1 identified a 36 kDa protein in homogenates from seven tumours displaying cytoplasmic only and one tumour demonstrating both nuclear and cytoplasmic immunostaining. Using restriction fragment length polymorphism polymerase chain reaction and PCR-multiplex analysis, amplification of the cyclin D1 gene (CCND1 was detected in 1/29 of the tumours demonstrating overexpression of cyclin D1 protein. We conclude that deregulation of CCND1 expression leading to both cytoplasmic and nuclear protein localization is a frequent event in ovarian cancer and occurs mainly in the absence of gene amplification.     

乳腺癌中RUNX3、cyclin D1和p27kip1蛋白的表达及其意义

[目的]探讨乳腺癌组织中RUNX3、cyclinD1和p27^kip1基因的表达及其临床意义。[方法]采用免疫组织化学SP法检测88例乳腺癌、40例乳腺纤维腺瘤、40例乳腺增生病中RUNX3、cyclinD1和p27^kip1的表达。[结果]（1）RUNX3和p27^kip1在乳腺癌中的阳性表达率分别为35．23％和51．14％，明显低于在乳腺纤维腺瘤（85％和82．5％）及乳腺增生病（87．5％和85％）中的表达率（P〈0．05）。乳腺癌中cyclinD1的表达率（63．63％）明显高于良性病变中表达。（2）RUNX3和p27^kip1蛋白表达与乳腺癌的临床分期、淋巴结转移呈负相关。p27^kip1蛋白表达与病理分级呈负相关。cyclinD1蛋白表达与病理分级呈正相关。（3）乳腺癌中RUNX3与p27^kip1蛋白的表达呈正相关性（P〈0．05）．与cyclinD1的表达呈负相关性（P〉0．05）。[结论]RUNX3、cyclinD1和p27^kip1在乳腺癌的发生发展中起重要作用。检测乳腺癌组织中RUNX3、cyclinD1和p27^kip1基因的表达对于评价患者的预后有一定价值。