ID-1、Ki-67及Bcl-2在食管鳞状细胞癌中的表达及意义

目的探讨食管鳞状细胞癌中DNA结合抑制蛋白-1（inhibitors of DNA binding1，ID-1）与Ki-67及Bcl-2表达的关系及其临床意义。方法应用免疫组织化学方法检测118例食管鳞状细胞癌手术切除标本及20例癌旁正常组织中ID-1、Ki-67及Bcl-2的表达情况。结果食管鳞状细胞癌中ID-1、Ki-67及Bcl-2表达上调，阳性比例分别为86．44％、81．36％和59．32％；ID-1及Bcl-2表达程度与肿瘤组织分化程度正相关（分别为r=0．289，P=0．002；及r=0．319，P=0．001）；Ki-67表达程度与肿瘤组织分化程度负相关（r=-0．320，P〈0．001）；ID-1与Bcl-2表达正相关而与Ki-67表达无关；ID-1、Ki-67及Bcl-2表达与患者的年龄、性别及是否伴淋巴结转移均无关。结论细胞凋亡抑制可能是ID-1参与食管鳞状细胞癌发生发展的主要机制，ID-1不适用于评价食管鳞状细胞癌的淋巴结转移。     

食管鳞癌组织中MICA基因mRNA、蛋白表达及意义

目的探讨MHCⅠ类链相关蛋白A（MICA）与食管鳞癌分化程度及淋巴结转移的相关性。方法采用RT-PCR法检测43例食管鳞癌组织（鳞癌组）和相应癌旁正常组织（癌旁组）中MICA mRNA表达,免疫组化SP法检测两组MICA蛋白表达,Spearman相关关系检验分析其相关性：结果鳞癌组MICA mRNA的表达水平显著高于癌旁组（t=8．35,P〈0．05）。鳞癌组MICA蛋白表达与癌组织分化程度及淋巴结转移有相关性,与肿瘤大小、性别、年龄无相关性（P〉0．05）;MICA蛋白表达水平与MICA mRNA表达水平呈显著正相关（rs=0．905,P〈0．01）。结论MICA高表达与食管鳞癌组织分化程度密切相关,可能在食管鳞癌的发生发展、浸润转移过程中起重要作用。     

Survivin和Ki67在食管鳞癌组织中的表达及意义

目的探讨Survivin和Ki67蛋白表达在食管鳞癌发生、发展中的作用。方法应用免疫组化SP法检测40例食管鳞癌患者癌组织中Survivin和Ki67蛋白表达情况,并分析其与临床特征的关系以及二者的相关性。结果食管鳞癌患者癌组织中Survivin和Ki67蛋白的阳性表达率分别为72．5％和75．0％。两种蛋白表达与肿瘤组织分化程度、浸润程度相关,而与性别、年龄、淋巴结转移情况无关。二者表达呈显著正相关（r=0．420,P〈0．05）。结论Survivin和Ki67蛋白与食管鳞癌组织的分化和恶性进展有关,Survivin在抑制细胞凋亡的同时可能促进了细胞增殖。     

Moesin、Radixin及Ki-67在子宫内膜样腺癌组织芯片中的表达及其临床意义

目的探讨Moesin、Radixin及Ki-67在子宫内膜样腺癌中的表达及其I临床意义。方法运用组织芯片技术及免疫组织化学sP法检测子宫内膜样腺癌组织中Moesin、Radixin和Ki-67的表达情况,并运用统计学方法分析染色结果与临床病理分期之间的关系。结果子宫内膜样腺癌组织中Moesin、Radixin及Ki-67的阳性表达率与正常子宫内膜组织相比差异均有统计学意义（P〈0．01）;Moesin和Radixin的表达与肿瘤浸润深度、淋巴结转移及远处转移显著相关（P〈0．05）,Ki-67的表达与肿瘤分化程度显著相关（P〈0．01）。Moesin和Radixin的表达具有相关性（P〈0．01）,二者的表达与Ki-67均无相关性（P〉0．05）。结论Moesin、Radixin和Ki-67的表达与子宫内膜样腺癌发生发展及侵袭转移有关,三者均可作为评价子宫内膜样癌的生物学行为的指标。     

口腔鳞癌组织中Cyclin D1、Ki-67基因的表达及意义

目的探讨口腔鳞癌组织中Cyclin D1、Ki-67基因蛋白表达与口腔鳞癌发生、发展的关系。方法取68份口腔鳞癌组织，用免疫组化法检测癌组织中Cyclin D1、Ki-67基因蛋白的表达，用积分PCR方法检测Cyclin D1的基因扩增。结果口腔鳞癌组织中Cyclin D1基因蛋白表达阳性36例(52．9％)，Ki-67基因蛋白表达阳性48例(70．5％)，Cyclin D1与Ki-67基因蛋白表达呈正相关(r=0．7861)，33份(48．5％)口腔鳞癌组织中Cyclin D1基因存在扩增，Cyclin D1基因扩增与Cyclin D1基因蛋白表达无相关性。Cyclin D1基因蛋白的表达与淋巴结转移有关，Ki-67基因蛋白的表达与肿瘤分化程度有关。结论Cyclin D1基因扩增及Ki-67基因蛋白表达水平可作为判断口腔鳞癌预后及其恶性程度的指标。     

CXCR4在食管鳞癌组织中的表达及临床意义

目的探讨趋化因子受体CXCR4在食管鳞癌组织中的表达及其与食管鳞癌发生、发展的关系。方法采用RT-PCR及Western blot法检测35例食管鳞癌及其癌旁对照组织中CXCR4的表达水平。结果CXCR4 mR-NA及蛋白在食管鳞癌组织中的表达水平显著高于癌旁对照组（P〈0．05）,癌组织中淋巴结转移组的CXCR4 mR-NA及蛋白表达显著高于无淋巴结转移组（P〈0．05）;癌组织中CXCR4蛋白在侵及外膜组高于未侵及外膜组（P〈0．05）,在Ⅲ期的表达高于Ⅱ期（P〈0．05）。结论趋化因子受体CXCR4与食管鳞癌的发生、侵袭和转移密切相关,可能成为抑制食管鳞癌侵袭转移的新靶点。     

食管鳞癌组织中NF-κB p65、VEGF的表达及其与血管密度测定的意义

采用免疫组化S-P法检测61例食管鳞癌患者组织中核转录因子-κB（NF-κB）p65和血管内皮生长因子（VEGF）的表达及CD105标记的肿瘤内徽血管密度（MVD）。结果NF-κB p65表达与食管鳞癌的组织学分级、浸润深度和淋巴结转移均呈正相关（P均〈0．01）,VEGF蛋白表达及MVD与食管鳞癌组织学分级和浸润深度均无关,但与淋巴结转移呈正相关（P〈0．05）;NF-κB p65与VEGF表达呈正相关（P〈0．01）;二者均与MVD呈正相关（P均〈0．05）。提示NF-κB p65参与食管鳞癌的发生、发展,其机制可能与VEGF的表达和肿瘤血管生成有关。     

5-脂氧合酶mRNA在食管鳞癌中的表达及与血管生成的关系

目的探讨5-脂氧合酶（5-LOX）和微血管密度（MVD）在食管鳞状上皮细胞癌（ESCC）组织中的表达及相关关系，及5-LOX与肿瘤浸润和转移的关系。方法用半定量RT-PCR法测定35例ESCC患者癌组织（T）和相邻癌旁组织（N）中5-LOX mRNA的表达，求得T／N值。同时用免疫组织化学法测定癌组织和癌旁正常组织中微血管密度（MVD）的T／N值。结果在35例ESCC患者中，33例5-LOX mRNA的T／N值〉1．0，占94．3％，34例MVD的T／N值〉1．0，占97．1％。5-LOX mRNA和MVD与食管鳞癌的肿瘤大小、浸润深度和淋巴结转移、肿瘤分化程度有关。5-LOX mRNA的T／N值与MVD的T／N值呈正相关。结论5-LOX与食管鳞癌血管生成和肿瘤浸润转移密切相关，其高表达可能促进肿瘤血管生成，从而促进ESCC的浸润和转移。     

血管紧张素Ⅱ及其受体1在食管鳞癌血管生成中的作用

目的 探讨血管紧张素Ⅱ（AngⅡ）及其受体1（AT1R）在食管鳞癌组织中的表达及其在血管生成中的作用。方法 应用免疫组化SP法检测30例食管鳞癌患者癌组织及其食管断端正常鳞状上皮中AngⅡ、AT1R和血管内皮生长因子（VEGF）蛋白的表达。结果 AngⅡ、AT1R和VEGF蛋白表达主要定位于鳞癌细胞iAngⅡ、AT1R在鳞癌组织中的表达均显著高于正常鳞状上皮（P均〈0．01）;癌组织中AngⅡ的表达程度与VEGF的表达程度呈正相关（ra′=0．4249,P〈0．05）,AT1R的表达程度与VEGF无相关性（ra′=0．1298,P〉0．05）。结论 食管鳞癌癌组织中高水平表达AngⅡ及AT1R,二者可能通过诱导VEGF的产生促进肿瘤新生血管的形成。     

胰岛素样生长因子结合蛋白-4在食管鳞癌中的表达及临床意义

目的探讨胰岛素样生长因子结合蛋白-4（IGFBP-4）在食管鳞癌中的表达及其临床意义。方法应用RT—PCR及免疫组化方法检测23例食管鳞癌及正常组织IGFBP-4、基质金属蛋白酶-7（MMP-7）的表达情况。结果食管鳞癌IGFBP-4 mRNA表达高于正常组织（P〈0．01），但在肿瘤侵及外膜以上及有淋巴结转移者表达降低（P〈0．01），在不同年龄、性别及肿瘤大小间表达差异无统计学意义（P〉0．05）。癌组织IGFBP-4蛋白表达阳性率为60．9％，低于正常组织（91．3％）（P〈0．05）。MMP-7 mRNA在食管鳞癌组织的表达为100．0％，正常组织未见表达（P〈0．01），肿瘤侵及外膜以上及有淋巴结转移组表达上调更明显（P〈0．05）。IGFBP-4与MMP-7 mRNA表达呈负相关（P〈0．01）。结论IGFBP-4表达下调与食管鳞癌的浸润及转移有密切关系，并与MMP-7高表达负相关，IGFBP-4 mRNA检测对判断食管癌进展及预后有重要意义。     

抑癌基因PTEN在人食管鳞癌中的表达及其意义

目的 研究食管鳞癌中PTEN的表达，探讨其与临床病理参数的关系及对患者预后的判断价值。方法 应用免疫组织化学S-P法检测62例食管鳞癌组织标本中PTEN的表达水平。结果 （1）PTEN的表达与食管壁浸润程度、淋巴结转移情况、远处转移情况、pTNM分期、肿瘤分化程度呈负相关关系（P〈0．05）。（2）高表达组患者的术后3年生存率明显低于低表达组，组间比较有统计学意义（P〈0．01）。结论 PTEN表达与反映食管鳞癌恶性度的临床病理参数呈负相关关系，是独立的预后评价因素之一。     

p53 negativity, CDC25B positivity, and metallothionein negativity are predictors of a response of esophageal squamous cell carcinoma to chemoradiotherapy

AIM: Esophageal squamous cell carcinoma is generally sensitive to chemoradiotherapy (CRT), but some cases are not. Using a retrospective analysis, we aimed to identify the predictors of the response by esophageal squamous cell carcinoma to definitive CRT. METHODS: The intensities of expression of,p53, Ki67, Bcl-2, Bax, cyclin D1, VEGF, CDC25B, and metalbthionein (MT) were evaluated immunohistochemically in the bfopsy specimens obtained before CRT, and the intensities of their expression were tested for correlations with the clinical effects of CRT. RESULTS: The esophageal squamous cell carcinomas with negative p53, positive CDC25B, and negative MT expression were found to be significantly more sensitive to CRT. In addition, p53 positivity and CDC25B positivity respomd well to CRT. CONCLUSION: Esophageal squamous cell carcinomas with negative p53,positive CDC25B, and negative MT expressions respond well to CRT. Even with p53 positivity, if with CDC25B positivity, CRT can be expected.     

The clinicopathological significance of p21 and p53 expression in esophageal squamous cell carcinoma: an analysis of 153 patients

OBJECTIVE: The p21 gene is thought to play a central role in tumor suppression. The aim of this study was to examine the clinicopathological role of p21 and p53 in esophageal squamous cell carcinomas. METHODS: The expression of p21 and p53 proteins in 153 Chinese patients (131 men, 22 women) with resected esophageal squamous cell carcinomas was investigated by the immunohistochemical method. Correlation between p21 and p53 expression and clinicopathological features was examined. RESULTS: The expression of p21 and p53 was detected in 70% and 64% of the tumors, respectively. The staining of p21 and p53 was also found in squamous carcinoma in situ, dysplasia, and nontumor epithelium. p21 expression was often weak in the suprabasal cells and found in better differentiated tumors. There was no significant correlation between the expression of p21 and the abnormal accumulation of p53. The prognosis of the patients depended on the size, stage, and p21 expression of the lesion. In stage III lesions with tumor diameter < or = 7.5 cm (n = 93), patients with loss of p21 expression had better survival. The survival rates of patients were worse if they had expression of both p21 and p53. CONCLUSIONS: Thus, p21 and p53 had prognostic value for esophageal squamous cell carcinomas. Loss of p21 expression was shown without p53 alternations, indicating that other mechanisms are also involved in turning off the gene. The pattern of p21 and p53 expression predicts an aggressive clinical course of esophageal squamous cell carcinomas.     

Clathrin heavy chain is a useful immunohistochemical marker for esophageal squamous intraepithelial neoplasia

Background

Recent advances in the endoscopic diagnosis and treatment of esophageal cancer have facilitated the detection and treatment of minute tumors, necessitating the accurate histopathological diagnosis of early esophageal cancer or precancerous lesions. This study evaluated the usefulness of immunohistochemical analysis (IHC) of clathrin heavy chain (CHC) as a marker for early esophageal cancer.

Methods

The immunoreactivity of CHC was analyzed in 409 esophageal specimens using a tissue array. Immunoreactivities of CHC, p53, and Ki67 were then compared in 44 endoscopically resected specimens.

Results

CHC expression was significantly stronger in the cytoplasm of esophageal squamous cell carcinomas compared with non-tumor specimens in the tissue array. CHC expression in endoscopic specimens was significantly stronger in the cytoplasm of high-grade intraepithelial neoplasias and superficial carcinomas than in benign squamous epithelium and low-grade intraepithelial neoplasias. The sensitivity and specificity of CHC for the diagnosis of esophageal lesions were 75 and 96 %, respectively. These accuracies were comparable with those of p53 (43 and 98 %) and Ki67 (68 and 100 %). In addition, the sensitivity was increased by using a combination of markers as follows: 80 %, CHC + p53; 78 %, CHC + Ki67; 90 %, CHC + p53 + Ki67.

Conclusions

CHC detected by IHC may be a useful marker for the pathological diagnosis of esophageal squamous intraepithelial neoplasia.     

Effect of estrogen on growth and apoptosis in esophageal adenocarcinoma cells

The epidemiology of esophageal adenocarcinoma demonstrates a strong gender bias with a sex ratio of 8–9:1 in favor of males. A potential explanation for this is that estrogen might protect against esophageal adenocarcinoma. Estrogen has previously been shown to stimulate apoptosis in esophageal squamous cancer cells. However, the effect of estrogen on esophageal adenocarcinoma cells has not been determined. We used immunoblotting analysis to determine the expression of estrogen receptors, cell adhesion marker E‐cadherin, and proliferation marker Ki‐67 in cell lines derived from esophageal adenocarcinoma (OE‐19, OE‐33) and Barrett's esophagus (QhTRT, ChTRT, GihTRT). Estrogen and selective estrogen receptor modulator (SERM)‐dependent effects on cell growth were determined by the CellTiter‐96 Aqueous Proliferation Assay. Apoptosis was determined by Annexin V/Propidium Iodide cell labeling and flow cytometry. We detected that physiological and supra‐physiological concentrations of 17β‐estradiol and SERM decreased cell growth in esophageal adenocarcinoma cells. In Barrett's esophagus cells (QhTRT, ChTRT), decreased growth was also detected in response to estrogen/SERM. The level of estrogen receptor expression in the cell lines correlated with the level of anti‐growth effects induced by the receptor agonists. Flow cytometry analysis confirmed estrogen/SERM stimulated apoptosis in esophageal adenocarcinoma cells. Estrogen/SERM treatments were associated with a decrease in the expression of Ki‐67 and an increase in E‐cadherin expression in esophageal adenocarcinoma cells. This study suggests that esophageal adenocarcinoma and Barrett's esophagus cells respond to treatment with selective estrogen receptor ligands, resulting in decreased cell growth and apoptosis. Further research to explore potential therapeutic applications is warranted.     

Carcinoma of the esophagus with mixed basaloid squamous and glandular differentiation: a distinct histological presentation

Esophageal cancer is the third most common gastrointestinal cancer and ranks among the ten commonest cancers worldwide. Histologically, approx 60% of esophageal cancers are adenocarcinomas and 40% are squamous cell carcinomas (SCC). Other rare cancers of the esophagus include small-cell carcinomas, squamous cell carcinomas with sarcomatous features, adenoid cystic carcinomas, and mucoepidermoid carcinomas. Basaloid squamous cell carcinoma (BSCC) or basaloid squamous carcinoma (BSC) is a distinct clinicopathological entity, seen more frequently in elderly males. Stage at presentation is often advanced and regional adenopathy or distant metastases are not uncommon at presentation. We describe an unusual case report of esophageal BSCC with glandular differentiation. The clinical significance of glandular differentiation in this rare type of tumor is not known.     

Determination of proliferative activity using Ki67 and expression of p53 in colorectal cancer

BACKGROUNDS AND PURPOSE: Classically, neoplasia has been considered a disturbance in the regulation of proliferation. In the search of a prognostic discriminant, the distribution of Ki67 as a marker of cell proliferation and the expression of p53 was studied in patients with colorectal cancer, to see whether this marker correlate with clinicopathological aspects. PATIENTS AND METHODS: Formalin-fixed paraffin-embedded tissues from 38 surgically resected adenocarcinomas of colon and rectal (5 Dukes A, 15 B and 18 C) were examined for p53 and Ki67 reactivity by immunohistochemistry (Dako). Nuclear staining of 5% of the cells was the criterion for a positive p53 reaction. For the study of Ki67 10 or more fields were chosen until 1000 nuclei per specimen were counted. The Ki67 labelling indices was determined by the number of positive nuclei/1000 cells. Correlation was done with the Dukes stage, grade of differentiation and percentual of recurrence. RESULTS: The mean Ki67 labelling indices was 40. The p53 was positive in 79% of the patients. The mean of Ki67-Li was higher in the positive p53 patients. However we did not lymphatic find differences between Ki67 or p53 expression and sex, age, Dukes stage, grade of differentiation, neural, venous or lymphatic invasion or recurrence. CONCLUSION: Ki67 and p53 could not be correlated to clinical and pathologic parameters in this study and was of limited use as a prognostic discriminant. The positive p53 cancers have more proliferative activity detected by a higher index of Ki67.     

p62 Expression in primary carcinomas of the digestive system

AIM: To characterize p62 expression and define the relationship between p62 expression and cell proliferation in primary carcinomas of the digestive system. METHODS: p62 expression was characterized in surgically resected tumor specimens from 60 patients with primary carcinomas of the digestive tract (including 22 esophageal carcinomas, 17 gastric carcinomas, and 21 colorectal carcinomas) and 40 patients with hepatocellular carcinoma (HCC) by immunohistochemistry (IHC). The cell proliferation was determined by IHC of Ki-67 in 40 patients with HCC. RESULTS: Twenty-two cases of esophageal carcinoma were histopathologically diagnosed as squamous cell carcinoma. We combined the gastric and colorectal carcinomas based on the equivalent histology. The 38 tumors in the combined groups, consisted of 17 well-differentiated, eight moderately differentiated, nine poorly differentiated carcinomas, and four mucinous adenocarcinomas. According to the criteria of Edmondson and Steiner, 40 patients with HCC were graded (2 grade Ⅰ,17 grade Ⅱ and 21 grade Ⅲ). p62 expression in primary carcinomas of the gastrointestinal tract (60/60,100%) was higher than that (27/40, 67.5%) of HCC (P<0.01, %2 = 19.63). High expression levels of p62 were positively correlated with histological grades in gastric and colorectal carcinomas (P<0.0001) and inversely associated with those in HCC (P = 0.0322). No significant correlations were observed for esophageal carcinomas (P = 0.8246). p62 expression was also detected in the cytoplasm of morphologically normal columnar epithelial cells adjacent to the cancer foci of gastric and colorectal carcinomas. In 40 HCC specimens, the mean Ki-67 labeling index (LI) was (19.6±16.0)%. It was (28.3±18.73)% in 12 cases with high p62 expression (+++), (7.53±14.83)% in 13 cases without p62 expression(-). Patients with a high p62 expression showed a significantly higher level of Ki-67 staining than those without p62 expression (P<0.05, t = 2,069). CONCLUSION: p62 expression is common in carcinomas of the digestive system and higher in carcinomas of the gastrointestinal tract than in primary HCC. p62 is a cellular differentiation-related protein. Cancer cells with a high p62 expression exhibited highgrowth fractions in HCC.     

Clinicopathological significance of cyclooxygenase‐2 and cell cycle‐regulatory proteins expression in patients with esophageal squamous cell carcinoma

The aim of this study was to examine the expression of the molecular markers cyclooxygenase‐2 (COX‐2), Ki‐67, cyclin A, and p27 in patients with esophageal squamous cell carcinoma (ESCC), to ascertain the relationship of these makers with the clinicopathological significance of the patients, and to assess the additional prognostic value of the expression profile of these proteins for ESCC patients. The expression levels of COX‐2, Ki‐67, cyclin A, and p27 proteins of a series of primarily resected ESCC samples were determined by immunohistochemistry method. Clinicopathological and molecular factors affecting survival were analyzed by multivariate analysis. A total of 78 specimens were included in this study. Expression of COX‐2 was observed in 43 (55.1%) cases, and high levels of expression of Ki‐67, p27, and cyclin A were observed in 57 (73.0%), 33 (42.3%), 43 (55.1%) cases, respectively. The results of univariate survival analysis indicated that more advanced tumor stage, lymph node involvement, systemic dissemination, the levels of expression of COX‐2, Ki‐67, cyclin A, and p27 were associated with survival (all P‐value < 0.05). Multifactorial survival analysis revealed that only lymph node involvement, over‐expression of cyclin A, and low p27 expression were associated with the survival of the patients (hazard ratios = 2.83, 4.7, 2.9, respectively; P= 0.025, 0.042, 0.005, respectively). Among the molecular markers assessed, the expression of cell proliferation markers cyclin A and p27 are independent prognostic factors in patients with ESCC, whereas neither COX‐2 nor Ki‐67 is of independent prognostic value.     

Nonmuscle myosin IIA is associated with poor prognosis of esophageal squamous cancer

Nonmuscle myosin IIA (myosin IIA) is a force-producing protein involved in the process of cell migration. Its expression has been considered as a bad prognostic indicator in stage I lung adenocarcinoma. However, the expression and clinical significance of myosin IIA in esophageal cancer has not been explored. In this study, we investigate the expression level of myosin IIA in 50 esophageal squamous cancer and 30 adjacent normal esophageal tissues by immunohistochemical staining and correlated its expression with clinicopathological features. Myosin IIA was expressed in all esophageal squamous cancer tissues (100%) and 8 of 30 adjacent normal tissues (26.7%, P = 0.000). In cancer tissues, elevated myosin IIA expression level was significantly correlated with increasing metastatic lymph nodes, poorer cancer differentiation, and advanced tumor stage. Further univariate analysis suggested that strong myosin IIA expression was associated with a significantly shorter overall survival (P = 0.021). In addition, MYH9 SiRNA was transfected into esophageal squamous cancer cell line (KYSE-510) to study the role of myosin IIA in cell migration. SiRNA-mediated depletion of myosin IIA in KYSE-510 cells significantly increased cell-matrix adhesion and attenuated cell migration ability (P = 0.000). In conclusion, these findings indicate that overexpression of myosin IIA may contribute to the progression and poor prognosis of esophageal squamous cancer, and this effect may be associated with increased cancer cell migration.