诱导型NOS与p53、Bax蛋白在胆囊良恶性病变中的表达及相关意义

目的　探讨诱导型一氧化氮合酶 (iNOS)与凋亡诱导基因p5 3、Bax蛋白在胆囊良恶性病变中的表达及相关意义。　方法　采用免疫组织化学SP法检测iNOS及p5 3、Bax蛋白在 16例慢性胆囊炎、11例慢性胆囊炎伴胆囊腺肌瘤增生及 2 4例胆囊腺癌中的表达。　结果　 1 在胆囊良恶性病变的胆囊壁中均有iNOS、Bax表达 ,在胆囊腺癌中iNOS、Bax表达下降 (P <0 0 5 ) ,p5 3仅在部分胆囊腺癌细胞核中有较强阳性表达 ;2 在胆囊良恶性病变中 ,iNOS与Bax表达呈正相关 (P <0 0 1) ,p5 3与Bax表达呈负相关 (P <0 0 1)。　结论　慢性胆囊炎尤其是慢性胆囊炎伴腺肌瘤增生是胆囊腺癌重要的危险因素 ,NO是重要中介分子 ,NO诱导胆囊良性病变恶变的作用与细胞凋亡有密切关系。     

胆囊良恶性病变Skp2和E2F1表达及其临床病理意义

目的研究胆囊腺癌、癌旁组织和慢性胆囊炎组织中Skp2和E2F1表达及其临床病理意义。方法108例胆囊腺癌、46例癌旁组织和35例慢性胆囊炎组织常规制作石蜡包埋切片,Skp2和E2F1染色采用免疫组化SP法。结果胆囊腺癌Skp2和E2F1表达阳性率及其评分明显高于癌旁组织和慢性胆囊炎（P〈0.01）;腺瘤癌变或高分化腺癌、肿块最大径〈2 cm、无淋巴结转移及未侵犯周围组织的病例Skp2和E2F1表达阳性率及其评分明显低于低分化腺癌或黏液腺癌、肿块最大径≥2 cm、淋巴结转移及侵犯周围组织病例（P〈0.05或P〈0.01）;胆囊癌中Skp2和E2F1表达呈密切正相关（r=0.56,P〈0.01）。结论Skp2和E2F1表达可能是反映胆囊癌发生、进展、生物学行为和预后的重要生物学标记物。     

胆囊良恶性病变组织中HMGA2和CD9表达及意义

目的 研究胆囊良恶性病变组织中HMGA2和CD9表达水平及其临床病理意义.方法 108例胆囊腺癌、46例癌旁组织、15例腺瘤和35例慢性胆囊炎手术切除标本常规制作石蜡包埋切片, HMGA2和CD9染色方法为EnVision免疫组化法.结果 胆囊腺癌HMGA2表达阳性率明显高于癌旁组织(χ2=16.13,P<0.01)、腺瘤性息肉(χ2=8.19,P<0.01)和慢性胆囊炎(χ2=21.41,P<0.01); 胆囊腺癌CD9表达阳性率明显低于癌旁组织(χ2=8.76,P<0.01)、腺瘤性息肉(χ2=3.96,P<0.05)和慢性胆囊炎(χ2=14.31,P<0.01);HMGA2表达阳性和(或)CD9表达阴性的良性病例的胆囊上皮均呈中至重度不典型增生.高分化、肿块最大径<2 cm、无淋巴结转移、未侵犯周围组织的病例HMGA2表达阳性率明显低于低分化、肿块最大径≥2 cm、淋巴结转移和侵犯周围组织的病例(P<0.05或P<0.01);高分化腺癌、肿块最大径<2 cm、无淋巴结转移和未侵犯周围组织的病例CD9表达阳性率明显高于低分化腺癌、肿块最大径≥2 cm、淋巴结转移和侵犯周围组织病例(P<0.05或P<0.01).经Kaplan-Meier生存分析发现HMGA2表达阳性病例术后生存期明显低于阴性表达病例(P=0.020),但CD9表达阳性病例术后生存期明显高于阴性表达病例(P=0.019);Cox多变量回归分析显示HMGA2阳性表达和(或)CD9表达阴性是反映胆囊腺癌预后不良的一个重要指标.结论 HMGA2和CD9表达与胆囊腺癌发生、临床生物学行为及预后有密切关系.     

胆囊腺上皮异型增生的层粘蛋白和表皮生长因？…

目的：探讨慢性胆囊炎所致腺上皮增生，异型，癌变过程中层粘层粘蛋白和表皮生长因子的表达及其关系，方法；对９９例慢性胆囊炎壁肥厚和１３例慢性胆囊炎肥厚型腺癌，采用化学Ｓ－Ｐ法，观察层粘蛋白和表皮生长的表达。结果：生胆囊炎引起胆囊壁结节状增厚，腺体增生，异型，最后导致壁肥厚型腺癌。     

胆囊腺上皮异型增生的层粘蛋白和表皮生长因子表达

目的：探讨慢性胆囊炎所致腺上皮增生、异型、癌变过程中层粘蛋白和表皮生长因子的表达及其关系。方法：对９９例慢性胆囊炎壁肥厚和１３例慢性胆囊炎后壁肥厚型腺癌，采用免疫组织化学ＳＰ法，观察层粘蛋白和表皮生长因子的表达。结果：慢性胆囊炎引起胆囊壁结节状增厚，腺体增生、异型，最后导致壁肥厚型腺癌。胆囊炎后壁腺上皮异型增生中层粘蛋白显示基底膜线性结构缺损，厚薄不匀；表皮生长因子表达腺上皮细胞随异型增生程度的增加而阳性表达递增。结论：慢性胆囊炎引起囊壁弥漫性或结节状增厚，分散其间的腺上皮细胞发生增生、异型，最后导致慢性胆囊炎后壁肥厚型腺癌；层粘蛋白和表皮生长因子有助于癌前病变的诊断和鉴别。     

Survivin和APC蛋白在原发性胆囊癌中的表达及意义

目的：研究survivin和APC蛋白在原发性胆囊癌中的表达情况，并分析其与病理特征可能关系。方法：收集胆囊癌标本50例、胆囊腺瘤20例、慢性胆囊炎20例。应用免疫组织化学方法检测survivin蛋白和APC蛋白的表达情况，同时收集胆囊癌患者的病理资料，并将结果进行分析。结果：（1）survivin蛋白在胆囊癌、胆囊腺瘤及慢性胆囊炎组织中阳性表达率分别为64％（32／50）、20％（4／20）和10％（2／20），差异有显著性（P〈0．05）。胆囊癌组与胆囊腺瘤组及胆囊炎组比较差异均具有显著性（P〈0．05）。Survivin阳性表达与病理资料均无明显相关性（P〉0．05）。（2）APC蛋白在胆囊癌、胆囊腺瘤及慢性胆囊炎组的阳性表达率分别为48％（24／50）、85％（17／20）及90％（18／20），差异有显著性（P〈0．05）。胆囊癌组阳性表达率显著低于胆囊腺瘤组和慢性胆囊炎组（P〈0．05）。APC蛋白的表达水平与患者的病理资料均无明显相关性（P〉0．05）。（3）Survivin阳性表达与APC蛋白表达呈明显负相关（rs=-0．530，P=0．000）。结论：胆囊癌组Survivin蛋白的表达显著高于胆囊腺瘤及胆囊炎组，说明凋亡可能在胆囊癌的发生中起重要作用；survivin和APC蛋白在胆囊癌中的表达呈明显的相关性，可能在胆囊癌的发生中起协同作用。     

胆囊腺癌神经浸润和神经生长因子表达与临床病理特征的关系

目的探讨胆囊腺癌组织中神经浸润及神经生长因子(nerve growth factor,NGF)的表达与胆囊腺癌临床病理特征的关系。方法收集82例胆囊腺癌标本的临床病理资料,采用免疫组化SP法检测82例胆囊腺癌组织中NGF的表达,分析胆囊腺癌的神经浸润与患者性别、组织分化、淋巴结转移及TNM分期的关系。结果 82例胆囊腺癌神经浸润的发生率为69. 51%。神经浸润与组织分化、TNM分期有相关性(P 0. 001);与患者性别、淋巴结转移无相关性(P0. 05)。82例胆囊腺癌NGF阳性率为76. 83%。NGF表达与神经浸润、组织分化程度、TNM分期有相关性(P 0. 05);与患者性别、淋巴结转移无相关性(P 0. 05)。结论胆囊腺癌的神经浸润与组织分化和TNM分期密切相关,提示发生神经浸润的胆囊腺癌可能有更大的转移风险,而NGF阳性可能对其有更好的提示作用。     

MUC5B、Villin、P53在胆囊黏膜幽门腺化生、肠上皮化生及胆囊腺癌中的表达及意义

目的：通过观察MUC5B、Villin、P53蛋白在胆囊黏膜幽门腺化生、肠上皮化生及胆囊腺癌的表达,探讨胆囊黏膜两种化生与胆囊腺癌发生的关系。方法：收集2013年1月至2015年1月兰州市第二人民医院病理科诊断的胆囊黏膜幽门腺化生40例、肠上皮化生40例及胆囊腺癌40例,采用免疫组化方法检测MUC5B、Villin、P53在胆囊黏膜幽门腺化生、肠上皮化生及胆囊腺癌的表达。结果：MUC5B在胆囊黏膜幽门腺化生、肠上皮化生、胆囊腺癌的阳性表达率分别为95.00%(38/40)、75.00%(30/40)、27.50%(11/40);Villin在三组中的阳性表达率分别为0.00%(0/40)、87.50%(35/40)、22.50%(9/40);P53在三组中的阳性表达率分别为2.50%(1/40)、7.50%(3/40)、80.00%(32/40)。各组间比较MUC5B在幽门腺及肠上皮化生的阳性表达率明显高于胆囊腺癌,差异有统计学意义(χ2=42.754,P=0.001);Villin在肠上皮化生的阳性表达率明显高于幽门腺化生及胆囊腺癌,差异有统计学意义(χ2=71.124, P=0.001);P53在胆囊腺癌中的阳性表达率明显高于幽门腺及肠上皮化生,差异有统计学意义(χ2=71.667,P=0.001)。MUC5B、Villin在幽门腺化生、肠上皮化生及胆囊腺癌的阳性表达率递减;P53在幽门腺化生、肠上皮化生及胆囊腺癌的阳性表达率递增。结论：幽门腺及肠上皮化生可能参与了胆囊腺癌的发生。     

大肠腺癌中iNOS和COX-2的表达及意义

目的 探讨大肠腺癌中iNOS和COX-2的表达及其与大肠腺癌生物学行为的关系.方法 采用SP免疫组化方法检测78例大肠腺癌标本及33例癌旁正常肠黏膜组织中的iNOS和COX-2的表达,并对两指标表达与临床病理参数之间进行相关分析.结果 ① iNOS和COX-2在大肠腺癌中阳性率分别显著高于正常大肠黏膜阳性率(P<0.05);② iNOS、COX-2的表达与大肠癌淋巴结转移明显相关(P<0.05);③ iNOS和COX-2在大肠腺癌中的阳性表达与临床分期有关,在TNM分期中Ⅲ+Ⅳ期阳性表达率高于Ⅰ+Ⅱ期(P<0.05);④ 大肠腺癌组织中COX-2表达与iNOS表达之间存在明显相关性(P<0.05).结论 COX-2和iNOS过度表达参与了大肠腺癌的发生发展过程,并与其淋巴结转移和TNM分期有关;大肠腺癌中COX-2和iNOS的表达具有正向协同性.     

胆囊癌组织P62蛋白的表达及其与血管生成关系

目的 :检测胆囊癌组织中P6 2蛋白、碱性成纤维细胞生长因子(bFGF)的表达和微血管密度 (MVD) ,探讨它们的相互关系 ,以及与胆囊癌临床病理特征的关系。方法 :4 1例胆囊癌和 2 2例慢性胆囊炎组织中 ,P6 2蛋白和bFGF的表达用SP免疫组化染色法进行检测。胆囊癌组织中MVD用抗CD34单抗 (mAb)做SP免疫组化染色进行检测。结果 :4 1例胆囊癌组织中 ,P6 2和bFGF表达阳性率分别为 6 3.4 %和 75 .6 % ,均高于 2 2例慢性胆囊炎组织 (P <0 .0 1)。P6 2的表达与胆囊癌淋巴结转移有关 (P <0 .0 5 ) ,与癌组织的分级、临床分期无显著关系。bFGF的表达与胆囊癌临床病理分期及组织学分级有关 (P <0 .0 5 ) ,而与淋巴结转移无显著关系。P6 2表达率与bFGF表达率呈正相关关系 (r=0 .5 2 1;P <0 .0 1)。两者的表达均与患者的年龄、性别、肿瘤的种类、是否伴有胆囊结石无关。胆囊癌组织中MVD明显高于慢性胆囊炎组织 (P <0 .0 1)。P6 2及bFGF表达阳性组织MVD高于表达阴性组织 (分别P <0 .0 5和P <0 .0 1)。MVD与胆囊癌的Nevin分期及淋巴结转移有关 (P<0 .0 1) ,与患者的年龄、性别、肿瘤种类、分化程度以及是否伴有胆囊结石无关。结论 :P6 2蛋白及bFGF的表达与胆囊癌组织中血管的生成相关 ,在胆囊癌的发生和发展过程中可能具有重     

Correlated expression of inducible nitric oxide synthase and P53, bax in benign and malignant diseased gallbladder

Our goal has been to investigate the expression and correlated significance of inducible nitric oxide synthase (iNOS) and P53, Bax in benign and malignant gallbladder diseases. We detected the expression of iNOS, P53 and Bax in the gallbladder wall by SP immunohistochemistry in 16 cases of chronic cholecystitis, 11 cases of chronic cholecystitis with adenomyoma and 24 cases of gallbladder adenocarcinoma. The percentage of positively marked tumor cells was counted under microscope and the intensity of immunoreactivity was graded. SPSS10.0 statistical software was applied for statistical analysis. In this study, we found that: (1) Both benign and malignant diseased gallbladder wall expressed iNOS and Bax. Compared to benign diseased gallbladders, their expression in adenocarcinoma was decreased (p < 0.05), P53 was expressed strongly only in nuclei of adenocarcinoma cells of some cases. (2) In benign and malignant diseased gall-bladders, iNOS expression was related positively to Bax (p < 0.01), the expression of P53 and Bax had a negative relationship (p < 0.01). The results suggested that both chronic cholecystitis and chronic cholecystitis with adenomyoma carry the risk of becoming malignant, especially the latter. NO is an important mediated molecule in cancer, there are intimate relationships between gallbladder cancer and apoptosis.     

Study of nucleolar organizer regions (NORs) in lesions of gallbladder

Tissue sections from 100 specimens of gallbladder including 40 cases of chronic cholecystitis, 30 cases of epithelial hyperplasia and 30 cases of well, moderately and poorly differentiated adenocarcinoma of gallbladder were studied for argyrophillic nucleolar organizer regions (AgNORs). The arithmetic mean +/- S.D. were calculated for each category. This showed NOR counts of chronic cholecystitis--(1.89+/-0.96), epithelial hyperplasia--(3.99+/-1.03), well differentiated adenocarcinoma (WDCA)--(7.04+/-1.34), moderately differentiated adenocarcinoma (MDCA)--(7.52+/-0.97) and poorly differentiated adenocarcinoma (PDCA)--(8.37+/-1.27). The study revealed increasing number of AgNORS from cases of chronic cholecystitis to epithelial hyperplasia to carcinoma gallbladder, but a considerable overlap in AgNOR counts of individual cases was observed suggesting that though AgNOR count cannot act as a specific diagnostic parameter for diagnosis of early carcinoma & dysplasia in isolated cases, they may prove to be a good adjunct to presently available imaging techniques and cytology.     

Clinicopathologic significance of minichromosome maintenance protein 2 and Tat-interacting protein 30 expression in benign and malignant lesions of the gallbladder

Gallbladder cancers are aggressive tumors with a poor prognosis and high mortality rate. To find specific biological markers for early diagnosis and prognosis and to develop possible alternative treatment strategies, we examined minichromosome maintenance protein 2 (MCM2) and Tat-interacting protein 30 (TIP30) expression in 108 gallbladder adenocarcinomas, 15 gallbladder polyps, 35 chronic cholecystitis tissues, and 46 peritumoral tissues using immunohistochemistry. Expression of MCM2 was significantly higher in adenocarcinomas than in peritumoral tissues (χ2 = 8.41; P < .01), adenomatous polyps (χ2 = 6.81; P < .01), and chronic cholecystitis (χ2 = 21.00; P < .01). In contrast, Tat-interacting protein 30 expression was significantly less in adenocarcinomas than in peritumoral tissues (χ2 = 13.26; P < .01), adenomatous polyps (χ2 = 4.76; P < .05), and chronic cholecystitis (χ2 = 18.93; P < .01). The benign lesions in gallbladder epithelium with positive MCM2 or negative Tat-interacting protein 30 expression showed moderate to severe atypical hyperplasia. Expression of MCM2 and absence of Tat-interacting protein 30 were significantly associated with poor differentiation, large tumor mass, lymph node metastasis, and invasion of adenocarcinoma. Univariate Kaplan-Meier analysis showed that either elevated MCM2 (P = .006) or lowered Tat-interacting protein 30 (P = .006) expression was closely associated with shorter overall survival. Multivariate Cox regression analysis revealed that expression of MCM2 (P = .007) or nonexpression of Tat-interacting protein 30 (P = .009) was an independent predictor of a poor prognosis in adenocarcinoma. Our results suggest that overexpression of MCM2 or loss of expression of Tat-interacting protein 30 is closely related to carcinogenesis, progression, biological behavior, and prognosis of gallbladder adenocarcinoma.     

Immunohistochemical detection of 8-hydroxydeoxyguanosine,a marker of oxidative DNA damage,in human chronic cholecystitis

Immunohistochemical detection of 8-hydroxydeoxyguanosine, a marker of oxidative DNA damage, in human chronic cholecystitis
Aims : Recent studies suggest that oxidative DNA damage induced during chronic inflammation may play a role in carcinogenesis in some organs. Although gallbladder carcinomas are frequently observed with a background of chronic cholecystitis, little is known about oxidative DNA damage in chronic cholecystitis. The aims of this study were to investigate the expression of 8-hydroxydeoxyguanosine (8-OHdG), a biomarker of oxidative DNA damage, in normal and chronically inflamed human gallbladder mucosa and compare its expression with clinicopathological findings.
Methods and results : 8-OHdG expression was immunohistochemically examined using a monoclonal antibody against 8-OHdG in human gallbladder specimens. In normal gallbladder ( n =5), no 8-OHdG expression was observed. In contrast, nuclear expression of 8-OHdG was detected in 28 of 31cases (90.3%) in gallbladder epithelial cells with chronic cholecystitis. The positive cells were predominantly observed in the areas of active inflammation with prominent cell infiltration. Quantitative analysis revealed that the number of 8-OHdG+ cells (labelling index) significantly ( r _s=0.671, P  < 0.05) correlated with the degree of the activity of mucosal inflammation, while gender, age, and the presence of gallstones did not influence the index.
Conclusions : Oxidative DNA damage is common in chronic cholecystitis, suggesting a possible link between chronic inflammation and gallbladder carcinogenesis.     

Expression of heat-shock protein gp96 in gallbladder cancer and its prognostic clinical significance

Purpose: To detect the expression and prognostic clinical significance of heat-shock protein gp96 (HSP gp96) in gallbladder cancer. Methods: Immunohistochemistry was used to detect and compare the rate of HSP gp96 expression in 107 samples of gallbladder cancer, 70 of gallbladder adenoma and 67 of chronic cholecystitis. The association of clinicopathological factors and patients’ survival were calculated by univariate and multivariate (Cox proportional hazard regression method) analysis. Results: The expression positive rate of HSP gp96 was 90.7% (97/107) in gallbladder cancer, 71.4% (50/70) in gallbladder adenoma and 47.76% (32/67) in chronic cholecystitis respectively. The positive rate of HSP gp96 in gallbladder cancer tissues was significantly higher than that in gallbladder adenoma and chronic cholecystitis tissues (P < 0.01). Multivariate and Cox regression analysis showed that positive of HSP gp96 (P = 0.026) expression was an independent poor prognostic predictor in gallbladder cancer. Conclusions: HSP gp96-positive expression is closely correlated with poor survival in gallbladder cancer.     

Enhanced cell kinetics, p53 accumulation and high p21WAF1 expression in chronic cholecystitis: comparison with background mucosa of gallbladder carcinomas

AIMS: Since neoplasia resulting from chronic inflammation has recently attracted increasing attention, we have investigated surgically removed gallbladders to examine the relationship between chronic cholecystitis and carcinogenesis. METHODS AND RESULTS: The mucosa of 108 cholecystectomy specimens without gallbladder cancer and 54 surgically resected gallbladder carcinomas were classified into three groups according to the degree of lymphocytic infiltration, and assessed immunohistochemically for Ki67, p53, p21WAF1 and apoptosis. In gallbladder mucosa without carcinoma, all four parameters tended to increase with the inflammation score (IS). Significantly positive correlations were revealed between Ki67 and p53, Ki67 and p21WAF1, and p53 and p21WAF1. However, in gallbladder carcinoma cases, values of p53 and p21WAF1 for background mucosa were elevated as compared to the mucosa of cholecystitis with low IS, but there was no correlation between their expression and IS, except for Ki67. CONCLUSIONS: Severe chronic cholecystitis is associated with acceleration of epithelial cell turnover, damaged cells being eliminated by apoptosis. The background mucosa of gallbladder carcinomas showed similar cell proliferative activity (Ki67) to that in cholecystitis, with no parallel changes of p53 and p21WAF1 expression, suggesting the possibility of unknown cofactors causing genomic damage.     

Microsatellite instability in chronic cholecystitis is indicative of an early stage in gallbladder carcinogenesis

The study of microsatellite instability (MSI) in cases of severe chronic cholecystitis and gallbladder carcinomas, to cast light on its significance for tumorigenesis, revealed MSI in 9 (30%) of 30 cases of cholecystitis and 7 (41%) of 17 carcinomas, respectively. In addition, 5 (33%) of 15 samples of background mucosa of carcinoma were positive. Respective figures for loss of heterozygosity were 3 (10%) of 30 cases of cholecystitis, 6 (35%) of 17 carcinomas, and 1 (7%) of 15 samples of adjacent nonneoplastic mucosa. No correlation was observed among MSI state, immunohistochemical hMLH1 or hMSH2 expression, and any clinicopathologic factors. MSI was observed not only in gallbladder tumors but also in severe chronic cholecystitis and background mucosa, suggesting that it may have an important role in early-stage gallbladder carcinogenesis.