Clinicopathologic significance of cyclooxygenase-2 overexpression in esophageal squamous cell carcinoma

BACKGROUND: Esophageal cancer is one of the most aggressive malignancies in the world, and whether multiple therapeutic modalities could improve long-term survival remains controversial. Recent studies have shown an increase of cyclooxygenase-2 (COX-2) expression in various malignancies, but its clinicopathologic role in esophageal squamous cell carcinoma (ESCC) remains unclear. METHODS: From 1993 to 1997, tissue samples from 96 patients with ESCC who underwent esophagectomy at our institution were collected for analysis. Cyclooxygenase-2 expression was examined by immunohistochemical staining, and further confirmed by Western blot analysis on six frozen tissues. Clinicopathologic data were analyzed to verify the significance. RESULTS: Cyclooxygenase-2 immunoreactivity was detected in 59 of 96 ESCC specimens (61%), and COX-2 overexpression (COX-2 high) was observed in 49% (47 of 96) of ESCCs. Statistical differences between COX-2 high and COX-2 low were found with respect to the status of distant metastasis (M factor) (p = 0.035) and tumor stage (p = 0.04). The survival was not significantly different between patients with and without COX-2 overexpression (p = 0.43). Using the Cox regression analysis, only the N factor (p = 0.0034) and M factor (p = 0.0325) were independent prognostic factors. CONCLUSIONS: Our results showed that in patients with ESCC, COX-2 overexpression was significantly correlated with fewer metastases and less advanced stage, but had no impact on survival. The status of local or distant lymph node metastasis was the most important prognostic factor. The biological role and pathophysiologic regulation of COX-2 overexpression in ESCC deserve further investigation.     

Expression profile of an androgen regulated prostate specific homeobox gene NKX3.1 in primary prostate cancer

PURPOSE: NKX3.1, a member of the family of homeobox genes, exhibits prostate tissue specific expression and appears to play a role in mouse prostate development. Rapid induction of NKX3.1 gene expression in response to androgens has also been described. On the basis of the established role of androgens in prostatic growth and differentiation and studies showing an association of aberrant homeobox gene expression with the neoplastic process, we hypothesize that alterations of NKX3.1 gene expression play a role in prostate tumorigenesis. MATERIALS AND METHODS: NKX3.1 expression was analyzed in matched, microdissected normal and tumor tissues from 52 primary prostate cancer specimens from radical prostatectomy by semiquantitative RT-PCR and in situ hybridization and correlated with the clinicopathologic features. NKX3.1 expression was quantified as differential expression between matched tumor and normal tissues and was grouped as overexpression in tumor tissue, reduced expression in tumor tissue and no change between tumor and normal tissues. Androgen regulation of NKX3.1 expression was also studied in LNCaP cells. Androgen receptor (AR) expression in prostate tumor and normal tissue was correlated with NKX3.1 expression. RESULTS: Comparison of NKX3.1 expression between normal and tumor tissues revealed overexpression in 31% tumor specimens (16 of 52), decreased expression in 21% tumor specimens (11 of 52) and no change in 48% specimens (25 of 52). When these expression patterns were stratified by organ confined and non-organ-confined tumor, a higher percentage of patients exhibited NKX3.1 overexpression in non-organ confined tumor (40%) versus organ confined tumor (22%). Elevated NKX3.1 expression significantly correlated with tumor volume and serum prostate specific antigen (PSA) level in the NKX3.1 overexpression group (p<0.05). Metastatic prostate cancer cell lines did not exhibit mutations in the protein coding sequence of NKX3.1. Additionally, the NKX3.1 expression correlated with AR expression (p<0.01) in vivo in human prostate tissues. Comparison of PSA and NKX3.1 expression in response to androgen revealed a rapid androgen mediated induction of NKX3.1 expression in LNCaP cells. In situ hybridization analysis of representative specimens confirmed RT-PCR observations. CONCLUSIONS: These results suggest an association of NKX3.1 with a more aggressive phenotype of carcinoma of the prostate. Correlation of AR expression with NKX3.1 in human prostate tissues underscores the androgen regulation of NKX3.1 in the physiologic context of human prostate tissues.     

PTEN和p53在食管鳞癌中的表达及意义

目的探讨抑癌基因PTEN、p53在食管鳞癌中的表达及意义。方法应用免疫组化S-P法检测食管鳞癌组织、癌旁正常食管组织中PTEN和p53的蛋白表达情况,分析其在食管鳞癌发生发展过程中的作用以及两者之间的作用关系。结果PTEN、p53在食管鳞癌组织中的阳性表达率分别是53.33%(32/60)、61.67%(37/60),对照组阳性表达率分别是91.67%(55/60)、3.33%(2/60)。PTEN阳性表达与食管鳞癌的分化程度、淋巴结转移、浸润深度、TNM分期有关(P<0.05),与年龄、性别无关(P>0.05)。p53阳性表达与食管鳞癌的分化程度、淋巴结转移、浸润深度有关(P<0.05),与TNM分期、年龄、性别无关(P>0.05)。PTEN与p53在食管鳞癌中的表达呈负相关关系(P<0.05)。结论PTEN低表达p、53高表达在食管鳞癌的发生发展中起重要的作用,二者相互作用,诱导食管鳞癌的发生发展。联合检测PTEN、p53对食管鳞癌的诊断、治疗有指导意义。     

食管鳞癌与细胞因子信号转导负调控因子3及其DNA甲基化的关系

目的 检测食管鳞癌(ESCC)组织中细胞因子信号转导负调控因子3(SOCS3)的DNA甲基化、mRNA及蛋白表达水平,探讨其在食管鳞癌发生、发展、浸润和转移中的作用.方法 采用甲基化特异性聚合酶链反应(MSP)、Real-Time聚合酶链反应(PCR)和Western blot法分别检测43例食管鳞癌组织中SOCS3的DNA甲基化、mRNA和蛋白表达水平,并与相应的癌旁正常食管组织进行对照研究,分析其与临床病理参数的关系.结果 (1)食管鳞癌组织SOCS3 DNA甲基化的阳性率(79.1%)明显高于癌旁组织(14.0%,P＜0.01);(2)食管鳞癌组织SOCS3 mRNA相对表达强度比值(0.53±0.30)明显低于癌旁组织(1.15±0.44,P＜0.01),食管鳞癌组织中甲基化组的SOCS3 mRNA表达(0.45±0.24)显著低于非甲基化组(0.86±0.29,P＜0.05);(3)食管鳞癌组织SOCS3蛋白表达(1.66±0.22)显著低于癌旁组织(1.83±0.15,P＜0.01),食管鳞癌组织中甲基化组SOCS3蛋白表达(1.61±0.21)显著低于非甲基化组(1.87±0.15,P＜0.01);(4)在TNM分期中Ⅲ期组表达均低于Ⅰ～Ⅱ期组(P＜0.05),伴有淋巴结转移组表达也都低于无淋巴结转移组(P＜0.05),未发现其在性别、年龄、家族史、吸烟史中有明显差异(P＞0.05);(5)食管鳞癌组织中SOCS3mRNA表达及其蛋白表达水平与肿瘤分化级别呈正相关(0.301＜r＜1,P＜0.05),与TNM分期、淋巴结转移呈负相关(-1＜r＜-0.301,P＜0.05).结论 食管鳞癌组织中SOCS3 DNA甲基化阳性率高,导致SOCS3基因表达下调,与食管鳞癌的分化、浸润和转移密切相关.     

STAT3及其磷酸化在食管鳞癌组织中的表达及其意义

目的 检测食管鳞癌组织中信号转导子和转录激活子3( STAT3)在mRNA、蛋白质和蛋白质磷酸化3种水平的表达,探讨其在食管鳞癌发生、发展、浸润、转移中的作用。方法 检测43例食管鳞癌组织中的STAT3 mRNA、STAT3和磷酸化STAT3( pSTAT3)的表达,并与相应癌旁正常食管组织作对照研究,分析STAT3 mRNA、STAT3和pSTAT3的表达与临床病理参数的关系。结果 43例实验样本中(1)食管鳞癌组织中STAT3 mRNA相对表达强度比值(1.43±0.59)较癌旁组织的比值(0.98±0.47)明显增高(P＜0.05);(2)食管鳞癌组织中STAT3、pSTAT3表达(2.16±0.39、1.40±0.15)也都显著高于癌旁组织(1.87±0.29、1.25±0.13,P＜0.05);(3)食管鳞癌组织中STAT3mRNA、STAT3和pSTAT3在肿瘤不同分化级别中表达差异有统计学意义,分化级别越低,表达水平越高(P＜0.05),并与肿瘤分化级别呈负相关(-1 ＜r＜-0.301,P＜0.05);它们在TNM分期中Ⅲ期组的表达均高于Ⅰ～Ⅱ期组(P＜0.05),伴有淋巴结转移组表达也都高于无淋巴结转移组(P＜0.05),并与两者呈正相关(两者均为0.301 ＜r＜1,P＜0.05);但未发现它们在性别、年龄、家族史、吸烟史中差异有统计学意义(P＞0.05)。结论 食管鳞癌组织中STAT3磷酸化异常激活后,导致STAT3mRNA、STAT3和pSTAT3的高表达,与食管鳞癌的分化、浸润、转移相关。     

Macrophage migration inhibitory factor stimulates angiogenic factor expression and correlates with differentiation and lymph node status in patients with esophageal squamous cell carcinoma

OBJECTIVE: The objectives of this study were: 1) to examine the expression of macrophage migration inhibitory factor (MIF) in esophageal squamous cell carcinoma (ESCC); 2) to see if a relationship exists between MIF expression, clinicopathologic features, and long-term prognosis; and 3) to ascertain the possible biologic function of MIF in angiogenesis. SUMMARY BACKGROUND DATA: MIF has been linked to fundamental processes such as those controlling cell proliferation, cell survival, angiogenesis, and tumor progression. Its role in ESCC, and the correlation of MIF expression and tumor pathologic features in patients, has not been elucidated. METHODS: The expression of MIF in tumor and nontumor tissues was examined by immunohistochemical staining. Concentrations of MIF, vascular endothelial growth factor (VEGF), and interleukin-8 (IL-8) in patients' sera and in the supernatant of tumor cells culture were examined by ELISA. Correlations with clinicopathologic factors were made. RESULTS: In 72 patients with ESCC, intracellular MIF was overexpressed in esophagectomy specimens. The expression of MIF correlated with both tumor differentiation and lymph node status. The median survival in the low-MIF expression group (<50% positively stained cancer cells on immunohistochemistry) and high expression group (>/=50% positively stained cancer cells) was 28.3 months and 15.8 months, respectively (P = 0.03). The 3-year survival rates for the 2 groups were 37.7% and 12.1%, respectively. MIF expression was related to microvessel density; increased MIF serum levels also correlated with higher serum levels of VEGF. In addition, in vitro MIF stimulation of esophageal cancer cell lines induced a dose-dependent increase in VEGF and IL-8 secretion. CONCLUSIONS: These results demonstrate, for the first time, that human esophageal carcinomas express and secrete large amounts of MIF. Through its effects on VEGF and IL-8, MIF may serve as an autocrine factor in angiogenesis and thus play an important role in the pathogenesis of ESCC.     

血管内皮生长因子D在食管鳞癌组织中的表达及其临床意义

目的探讨血管内皮生长因子D（VEGF．D）在食管鳞状细胞癌中的表达及意义。方法收集中山大学附属第一医院2009年3月至2010年2月间收治的39例食管鳞癌及癌旁组织标本。用原位杂交法检测肿瘤组织及癌旁组织中VEGF．DmRNA的表达情况;用免疫组织化学法检测D2．40表达,计算D2—40标记的微淋巴管密度（MLVD）。分析VEGF．DmRNA表达与食管鳞癌临床病理资料及MLVD的关系。结果肿瘤组织中VEGF．DmRNA阳性表达率为56.4％（22／39）．明显高于癌旁组织[2．6％（1／39）,P〈0．05]。有淋巴结转移者VEGF．DmRNA阳性表达率明显高于无淋巴结转移者E92．9％（13／14）比36．0％（9／25）,P〈0．05]。VEGF．DmRNA阳性表达者MLVD明显高于阴性表达者（8．20±1．22比5.31±0．97,P〈0．01）。结论VEGF—D可能通过诱导淋巴管生成促进食管鳞癌淋巴转移。     

Glypican-3 expression is markedly decreased in human gastric cancer but not in esophageal cancer

BACKGROUND: Deregulation of the expression of glypican-3, a heparan sulfate proteoglycan, has been demonstrated in several human cancers. METHODS: In the present study, glypican-3 mRNA expression was analyzed by Northern blotting and in situ hybridization in 20 normal and 41 cancerous esophageal specimens as well as in 15 normal and 32 cancerous gastric tissues. RESULTS: Glypican-3 mRNA was expressed in both normal and esophageal cancer tissues without a significant difference between normal and cancerous tissues, and without a correlation with histological type, tumor stage, tumor grade, or patient survival. Moderate to strong glypican-3 mRNA signals were found in the cytoplasm of squamous epithelial cells of the normal esophagus. In both squamous and adenocarcinomas of the esophagus glypican-3 mRNA signals were also moderately to strongly present in the cytoplasm of the cancer cells. In gastric tissues, glypican-3 mRNA was present in 53% of normal gastric tissue samples, but was below the detection level in all examined gastric cancer samples. Glypican-3 mRNA signals were moderately to strongly present in the cytoplasm of gastric mucosal epithelial cells, but were only very faintly present in some cancer cells. CONCLUSIONS: Glypican-3 may be involved in the growth control of normal esophageal and gastric epithelial cells. Furthermore, our results suggest that glypican-3 may play a tumor suppressor role in gastric but not in esophageal cancer.     

促肝细胞再生磷酸酶-3基因在结直肠癌组织中的表达及其临床意义

目的检测促肝细胞再生磷酸酶-3(PRL-3)mRNA在结直肠癌(CRC)中的表达,探讨其与CRC发生发展以及侵袭转移的关系。方法半定量PCR测定46例CRC组织及其对应正常黏膜、6例结直肠腺瘤(CRA)及其对应正常黏膜以及18例淋巴结和肝转移灶中PRL-3mRNA相对表达水平,并分析其表达水平与临床病理学指标的关系。单链多态性分析法(PCR-SSCP)检测基因突变情况。结果46例CRC中PRL-3基因表达量较相应正常黏膜组织高,差异有统计学意义(1.6±0.7比0.4±0.1,P<0.01);6例结直肠腺瘤组织与对应的正常黏膜中PRL-3mRNA表达量差异无统计学意义(0.6±0.3比0.5±0.2,P>0.05);12例淋巴结转移灶和6例肝转移灶PRL-3基因表达量分别为2.1±0.8和3.3±1.0,显著高于原发癌肿、正常黏膜、阴性淋巴结组织中的表达(P<0.01)。PRL-3基因表达水平与CRCDukes分期、浸润深度、淋巴结和远处转移有关(P<0.05),与性别、肿瘤大小、分化程度无关(P>0.05)。PCR-SSCP分析发现,6例肝转移灶中有1例出现异常泳动条带。结论PRL-3基因在CRC的发生发展和侵袭转移中起重要作用,其作用可能是通过基因表达上调和基因突变共同实现的。     

AURORA-A在肝恶性肿瘤中的表达及意义

目的　研究Aurora AmRNA在肝癌组织中的表达 ,探讨Aurora A基因与肝癌发生的关系。方法　采用半定量逆转录聚合酶链反应 (RT PCR)检测 30例肝癌组织及 30例正常肝组织中的Aurora AmRNA。结果　Aurora AmRNA在肝癌组织中的表达明显高于正常肝组织 (P <0 .0 5 )。结论　Aurora AmRNA在肝癌组织中的过度表达提示Aurora A基因与肝癌的发病及发展有关。     

RhoE与EGFR在食管鳞癌中表达的研究

目的：探讨RhoE与EGFR在食管鳞状细胞癌中的表达及其关联性,并分析其临床意义。方法运用快捷免疫组化方法对我院2011年1月至2012年1月收集的食管鳞状细胞癌癌组织及相应的癌旁组织59例进行检测,测定RhoE与EGFR的表达情况,运用统计学方法结合患者其他指标进行分析。结果①食管鳞癌癌旁正常组织中RhoE蛋白的阳性表达灰度值是52．131±3．682,食管鳞癌组织中为37．115±4．314,二者差异有统计学意义（P＜0．001）。②食管鳞癌癌旁正常组织中EGFR蛋白的阳性表达灰度值是33．956±3．716,食管鳞癌组织中为48．168±2．945,二者差异有统计学意义（P＜0．05）。③二者表达水平情况与肿瘤的临床分期、分化程度、淋巴转移以及有无浸润都有密切关系（P＜0．001）,与肿瘤的大体分型、大小和部位无关（P＞0．05）。④RhoE与EGFR在食管鳞癌组织中的表达呈负相关（P＜0．01）。结论 EGFR基因的表达水平在食管鳞癌中是增高的,而RhoE则是降低的;并且二者呈负相关,与肿瘤的分化程度、有无浸润、淋巴的转移以及临床分期等密切相关,这提示着EGFR的高表达或许参与了肿瘤的发生与发展,对二者的表达情况进行检测有助于我们了解食管癌的生物学行为以及评估预后。     

Reduced tumor vessel density and high expression of glucose transporter 1 suggest tumor hypoxia of squamous cell carcinoma of the esophagus surviving after radiotherapy

BACKGROUND: Squamous cell carcinoma of the esophagus (ESCC) is radiosensitive; however, surgeons frequently encounter ESCC that survives radiotherapy to grow more rapidly and invasively. This alteration of tumor behavior may result from tumor hypoxia induced by radiotherapy. METHODS: Forty-four patients with advanced (T3 and T4) ESCC, who underwent radiotherapy before operation, either with 40 Gy for preoperative treatment or 60 Gy or more for radical treatment, and 44 patients without preoperative therapy were subjected to retrospective immunohistochemical study. CD34 for tumor vessels, glucose transporter 1 (GLUT1) which was induced by hypoxia, MIB-1 for proliferating activity, and p53 were stained for surgical samples from ESCC patients. Tumor tissue at the invading front was the focus of evaluation. Macroscopic morphologic differences of ESCC were also evaluated. RESULTS: Loss of esophageal wall thickness and deep ulceration were morphologic characteristics of ESCC after radiotherapy. Tumor vessel density was reduced and GLUT1 expression was greater in the ESCC after radiotherapy than in those without treatment. Tumor vessel density was similar for both preoperative and radical radiotherapy samples, while GLUT1 expression tended to be greater in the latter than in the former. The expression of MIB-1 and p53 did not show any significant difference between ESCC with or without radiotherapy. CONCLUSIONS: Reduced vessel density and increased GLUT1 expression suggested tumor hypoxia for ESCC occurred after radiotherapy. Tumor hypoxia would induce ulcerative and invasive growth, which is a great obstacle to clinical treatment of residual or relapse ESCC after radiotherapy.     

骨桥蛋白在大肠癌和大肠癌肝转移灶中的表达及其临床意义

目的：检测骨桥蛋白（osteopontin OPN)mRNA在大肠癌及大肠癌肝转移灶中的表达,并探讨其在大肠癌发生、发展及肝转移中的临床意义。方法：提取44例大肠癌及癌旁正常组织,20例大肠癌肝转移灶及3例正常肝组织RNA,采用RT－PCR半定量法检测其OPN mRNA的表达。并用原位杂交行组织定位检测。结果：44例大肠癌组织OPN表达高于癌旁正常组织（t=4.89,P=0.000)。44例大肠癌组织OPN mRNA表达率为18．2％,而在20例肝转移灶中表达率达80．0％（χ^2=22.42,P=0.000）。原位杂交表明,OPN mRNA定位于大肠癌细胞浆。结论：OPN mRNA在大肠癌组织中表达高于正常组织,而在大肠癌肝转移灶中呈更高表达,提示OPN可能是大肠癌肝转移的预测指标之一。     

Pin1和周期素D1在胰腺癌中的表达及其意义

目的:检测胰腺癌及癌旁组织中Pin1和周期素D1基因的表达,探讨Pin1在胰腺癌发病中所起的作用。方法:收集27例胰腺肿瘤组织及其相应的肿瘤旁组织标本,采用实时荧光定量逆转录聚合酶链反应法（RQ RT-PCR）检测胰腺良恶性肿瘤及肿瘤旁组织中Pin1和周期素D1 mRNA 的表达,运用Fisher精确概率分析两者之间的相关性及其与肿瘤临床分期和病理特征的关系。结果:7例胰腺囊腺瘤中周期素D1和Pin1的表达与肿瘤旁组织之间无显著性差异;而20例胰腺癌中周期素D1和Pin1的表达明显高于肿瘤旁组织［（2.78±1.02）vs.（4.36±1.27）和(5.48±1.69) vs. (9.97±1.86),P<0.05）］。Pin1和周期素D1与肿瘤的临床分期和病理分化程度无明显的相关（组间差异均为P>0.05）,但Pin1与周期素D1的表达有关（P<0.01）。 结论:胰腺癌中Pin1的过表达可促进周期素D1表达,由此诱导了肿瘤的发生;Pin1可能在胰腺癌中起着重要作用。