胃窦癌组织中MMP-26 mRNA及蛋白的表达

目的探讨胃窦癌及癌旁正常胃黏膜组织中MMP-26 mRNA及蛋白的表达意义。方法应用RT-PCR检测40对胃窦癌及癌旁正常胃黏膜组织中MMP-26基因的表达,Western blot方法检测其中10对标本的MMP-26蛋白表达,同时采用免疫组化MaxVision法检测102例胃窦癌组织和58例癌旁胃黏膜组织MMP-26蛋白的表达,分析MMP-26的表达与临床病理变量之间的关系。结果 MMP-26 mRNA在27例胃窦癌中的表达明显高于正常胃黏膜(67.5%)。Western blot结果显示7例胃癌组织中MMP-26蛋白表达明显高于对应的正常胃黏膜组织。免疫组化染色结果显示102例胃癌组织中有57例(55.9%)对MMP-26呈阳性反应,癌旁胃黏膜组织58例中有8例阳性(13.8%)。且MMP-26蛋白表达与胃癌浸润深度、淋巴结转移及TNM分期呈正相关。结论 MMP-26蛋白表达与胃癌浸润转移密切相关,可能作为胃癌早期诊断及预测预后的分子标记物。     

胃癌组织IL-17RB的表达及意义

分析胃癌组织IL-17RB和IL-17B表达水平,探讨其与胃癌发生发展的关系及其可能机制。留取25例胃癌组织及其配对的癌旁组织,应用普通PCR和实时荧光定量PCR法检测IL-17RB和IL-17B的mRNA,采用Pearson法进行相关性分析;western blot和免疫荧光法检测胃癌组织及其配对的癌旁组织IL-17RB蛋白水平表达差异。结果显示,胃癌组织IL-17RB和IL-17B mRNA表达水平均明显高于癌旁组织,两者呈明显正相关;胃癌组织IL-17RB蛋白水平明显高于癌旁组织。在胃癌组织,IL-17B/IL-17RB信号对胃癌的发生发展具有一定作用。     

IL-8在胃癌组织上皮间质转化过程中表达增强

目的研究胃癌上皮间质转化(EMT)过程中趋化因子IL-8的调节机制与意义。方法用Western blot、酶联免疫吸附试验(ELISA)等技术,检测胃癌组织与正常组织中IL-8以及EMT相关基因(E-cadherin, vimentin, twist等)的表达,胃癌细胞系MGC-803对IL-8与胃癌细胞EMT的相关性进行分析和探讨。结果肿瘤组织中IL-8的表达水平(710.5±89.9)显著高于癌旁组织(510.5±85.5)(P0.001)。与正常组织相比,胃癌组织中IL-6、twist及vimentin表达水平明显高于正常胃黏膜组织(P0.05),而E-cadherin表达水平低于正常组织(P0.05)。IL-8因子的分泌与twist蛋白(P0.001)和vimentin蛋白(r=0.454,P0.001)表达水平呈显著正相关性,与E-cadherin蛋白表达水平呈显著负相关性(P0.001)。结论胃癌组织中趋化因子IL-8的表达量增强, IL-8与胃癌侵袭转移相关,同时IL-8的表达量也可作为胃癌侵袭转移患者预后判断的参考标准之一。     

青海高原藏族胃癌患者自然杀伤细胞活化性受体D及其配体MICA表达的研究

目的： 研究高原藏族胃癌患者外周血NK细胞表面活化性受体D(NKG2D)的表达及局部肿瘤组织和癌旁正常组织中相应配体MICA 的表达,探讨宿主NKG2D 在抗胃癌中的作用及其与肿瘤免疫逃逸的关系。方法: 对33例高原藏族胃癌患者及20 例健康人,采用流式细胞术检测外周血NKG2D 的表达状况,RT-PCR 和免疫组织化学技术检测在局部肿瘤组织和癌旁正常组织标本中MICA 的表达。结果: 高原藏族胃癌患者外周血NKG2D 的表达为(13.47 ±5.26)%，明显低于正常组（32.62±10.08)%，2组之间差异显著(P<0．05); 藏族胃癌组织MICA mRNA的表达水平显著高于癌旁正常组织（P<0.05）。胃癌组织MICA蛋白表达在癌旁正常组织中为(21.21%,7/33),胃癌组织阳性率为(78.79%，26/33),高分化组(60.00%,3/5),中分化组(72.73%,8/11),低分化组(88.24%,15/17),组间差异显著(P<0.05),而与肿瘤的大小、性别、年龄、淋巴结转移无关（P>0.05）。 Spearman相关分析显示，MICA表达水平与mRNA的表达水平显著正相关（r=0.903,P<0.01）。结论: 高原藏族胃癌的免疫逃逸可能与NKG2D 表达下调及其配体MICA 的表达升高有关，高原藏族胃癌患者外周血NK细胞活性降低,其NKG2D 表达的下降是NK细胞活性下降的原因之一。NKG2D配体MICA 的基因表达可能与高原胃癌患者的恶性转化有一定的相关性。     

抗凋亡基因bcl—2在胃癌和癌旁组织中的表达及其临床意义

通过抗凋亡蛋白bcl-2在胃癌、癌旁组织中的表达与分布，探讨bcl-2基因在胃癌恶性转化过程中的作用机制和临床意义。应用S－P免疫组化法检测33例胃癌、17例癌旁和13例正常胃粘膜中bcl-2蛋白的表达。正常胃粘膜中bcl-2仅一例弱表达。在胃癌和癌旁bcl-2表达均高于正常胃粘膜（P＜0．05－0．01），但胃癌与癌旁间无差异。bcl-2表达与胃癌的分化程度，病理分期，淋巴结转移等病理特征无统计学差异。结果表明bcl-2基因参与了胃癌的发生。bcl-2蛋白在胃良性病变过度表达，一旦癌变，细胞的阳性表达不再升高，这可能与bcl-2抑制细胞凋亡的强度不再增加有关。     

Wntless在胃腺癌组织中表达及其意义

目的观察Wnt蛋白分泌相关的基因Wntless在胃癌腺癌细胞和癌旁正常黏膜细胞的表达,分析Wntless表达与胃腺癌病理指标之间相关性和临床意义。方法用免疫组化法检测胃腺癌组织和胃癌旁正常黏膜中Wntless的蛋白表达。结果在104例胃癌标本中,Wntless在胃腺癌组织中表达为:7.7%(8/104)例为0或1+、36.5%(38/104)例为2+、55.8%(58/104)例为3+;在癌旁正常黏膜表达为:71.2%(74/104)例为0或1+,19.2%(20/104)例为2+,9.6%(10/104)例为3+。Wntless在胃腺癌组织中的表达显著高于胃癌旁正常黏膜表达(P0.001)。Wntless蛋白表达与肿瘤分化呈正相关(P0.05),与淋巴结转移呈正相关(P0.05)。结论 Wntless蛋白在胃腺癌中高表达,可能对肿瘤发生和进展等发挥了促进作用,并可能为新的靶向治疗提供分子靶标。     

IL-17通过NF-κB信号通路参与胃癌发生发展的免疫调控

目的探讨白介素-17(IL-17)通过NF-κB信号通路参与胃癌发生的分子机制。方法选取甘肃省武威肿瘤医院胃癌患者30例及同一时间在医院体检的健康人30名;酶联免疫吸附法检测胃癌患者及健康人血清中IL-17、IL-6和IL-8的表达;用HE染色法观察胃癌患者胃组织病理学变化;免疫组织化学法检测胃癌患者的胃癌组织、癌旁组织及远离癌组织的胃组织中IL-17、NF-κB p65、IKK-β和IL-6的表达。Western blot检测胃癌患者的胃癌组织、癌旁组织及远离癌组织的胃组织IL-17、NF-κBp65、pNF-κB p65、IKK-β和IL-6的表达。结果与对照组相比,IL-17、IL-6和IL-8与在胃癌患者血清中表达上调(P0.05);IL-17、NF-κB p65、IKK-β和IL-6在胃癌组织中的表达明显高于正常及癌旁组织(P0.05)。与正常胃组织相比,胃癌组织中IL-17、NF-κB p65、pNF-κB p65、IKK-β和IL-6表达增加(P0.05);癌旁组织中pNF-κB p65、IKK-β和IL-6表达增加(P0.05)。与癌旁组织相比,胃癌组织中IL-17、NF-κB p65、pNF-κB p65、IKK-β和IL-6表达增加(P0.05)。结论 IL-17可能通过NF-κB信号通路刺激IL-6参与胃癌的免疫调控以及促进胃癌的发生发展,为胃癌发病机制提供新的见解。IL-17可能作为胃癌的临床治疗潜在靶点。     

E-cadherin基因在肺癌发生过程中的作用分析

目的检测E-cadherin在肺癌组织、癌旁组织及正常肺组织中的表达,并探讨其在肺癌发生发展中的作用。方法采用免疫组化SP法对22例肺癌组织、相应的癌旁组织和9例肺部良性病变旁正常肺组织中E-cadherin的表达状况进行检测。结果22例肺癌组织中E-cadherin蛋白表达减弱或缺失率59.1%(13/22)。明显高于相应癌旁组织中蛋白表达减弱或缺失率27.3%(6/22);9例正常肺组织中E-cadherin均正常表达。结论E-cadherin表达减弱或缺失是肺癌发生中的常见分子事件,可能参与了肺癌的发生发展过程。     

胃癌中YAP与β-catenin蛋白表达的临床意义及相关性

目的探讨YAP及β-catenin在胃癌组织、癌旁正常组织的表达与临床病理特征的关系及两者表达相关性。方法采用荧光定量PCR、免疫组化和Western blot测定YAP和β-catenin在50例胃癌组织及癌旁正常组织中mRNA及蛋白表达,并分析其与临床病理参数关系及两者表达的相关性。结果胃癌组织中YAP和β-catenin蛋白阳性表达率分别为72%(36/50)和64%(32/50),癌旁正常组织中阳性表达率分别为14%(7/50)和20%(10/50),P0.05;YAP及β-catenin mRNA和蛋白在胃癌组织中表达较正常组织高(P0.05),且两者表达水平与胃癌分化程度、淋巴结转移和临床分期有显著关系(P0.05);YAP与β-catenin在胃组织中共表达呈明显正相关(r=0.72,P0.05)。结论 YAP和β-catenin蛋白在胃癌组织中存在高表达共存现象,可能与胃癌细胞恶性程度及细胞分化有关,参与胃癌的发生、发展过程。     

Expression of RORα in gastric cancer and clinical pathological significance

目的 观察维甲酸相关孤核受体α(Retinoid acid receptor related Orphan Receptor α,RORα)蛋白在胃癌中的表达,探讨其与胃癌发生、发展的关系,为寻找胃癌肿瘤标记物提供一定的实验依据.方法 应用组织芯片技术及免疫组化SP法检测90例胃癌组织、48例癌旁胃黏膜组织与22例正常胃黏膜组织中RORα的表达,研究RORα与胃癌发生、发展的关系.结果 RORα蛋白在胃癌组织中表达下调,正常胃黏膜、癌旁胃黏膜和胃癌组织中RORα蛋白阳性率分别为83.36%(19/22),56.25%(27/48)和24.44%(22/90),3者相比差异有显著性(P＜0.05).高分化腺癌、中分化腺癌、低分化腺癌、黏液腺癌和印戒细胞癌中,RORα阳性率分别为45.00%(9/20)、27.59%(8/29)、14.29%(3/21)、11.11%(1/9)和9.05%(1/11).高分化腺癌RORα阳性率高于低分化腺癌(P＜0.05).结论 RORα蛋白下调与胃癌的发生、发展相关,且可能与胃癌的分化程度有关.     

Analysis of the correlation between P27 expression and Helicobacter pylori infection in gastric cancer

We aimed to explore the correlation between P27 expression and Helicobacter pylori (H. pylori) infection in gastric cancer, so as to provide evidence for understanding the pathogenesis of gastric cancer caused by H. pylori infection. A total of 82 samples of gastric cancer tissues and 56 samples of tumor-adjacent normal tissues collected from the gastrectomy were enrolled in this study. Then, 14C-urease breathing test was carried out to evaluate the infection of H. pylori in gastric cancer tissues, the expression of P27 in the tissue samples was detected by the immunohistochemistry staining, and the correlation between the H. pylori infection and P27 expression in gastric cancer was analyzed. Of 82 gastric cancer patients, there were 53 patients with H. pylori infection (64.63%). Among the patients with highly or moderately differentiated gastric cancer, the expression of P27 was much higher than that of patients with poorly differentiated gastric cancer (p < 0.01). Besides, comparison of the P27 expression between males and females, among different age groups, tumor sizes, TNM stages, tumor infiltration degrees, or lymph node metastasis, showed no significant differences (p > 0.05). Analysis of the correlation revealed that P27 expression was negatively correlated with the infection of H. pylori (p < 0.01). Multifactorial logistics regression analysis indicated that tumor differentiation was a risk factor of P27-positive expression in gastric cancer tissues (p < 0.01). In addition, P27 expression in the gastric cancer tissues was lower than that in the tumor-adjacent normal tissues (p < 0.01). In gastric cancer patients, expression of P27 is correlated with H. pylori infection which, via downregulating P27, can cause the cancerization of gastric mucosa, and P27, for its role in the development and progression of gastric cancer, is a potential auxiliary indicator for clinical diagnosis whether gastric cancer is complicated with H. pylori infection. So, P27 is a key indicator for diagnosis, treatment, and prognostic evaluation of disease in the advanced stage.     

胃癌组织中microRNA的差异表达

目的 检测胃癌组织和癌旁组织中microRNA (miRNA)的差异表达.方法 收集经病理确诊为胃癌进行外科手术切除的25例患者的胃癌组织及距离胃癌病灶5 cm以上的癌旁组织,选取其中7例标本提取总RNA,应用Illumina miRNA芯片检测胃癌和癌旁组织中差异表达的miRNA.应用定量实时PCR技术验证25例胃癌和癌旁组织标本中差异表达的miRNA,并分析25例患者的临床病理与胃癌组织中miRNA差异表达的关联.结果 Illumina miRNA芯片检测显示,HS-138,HS-153,HS-157在胃癌组织中表达下调,miR-181a,miR-21,miR-21*,miR-27a,miR-584,miR-93在胃癌组织中表达上调.定量实时PCR显示,与癌旁组织比较,胃癌组织中miR-181a表达上调(56.848±135.551比3.950±12.101,P＜0.05),而miR-584在胃癌和癌旁组织中的表达差异无统计学意义(P＞0.05).胃癌组织miR-181a表达与患者胃癌淋巴结转移及年龄有关联(rp=0.462、0.414,P=0.009、0.023),与性别和病理分型无关联(P=0.220、0.106).结论 应用miRNA芯片技术和荧光定量PCR技术发现了胃癌组织中差异表达的miRNA.     

Expression of interleukin-8 correlates with vascularity in human gastric carcinomas.

Interleukin (IL)-8 is a multifunctional cytokine that can stimulate the division of endothelial cells. We examined the expression of IL-8 mRNA using Northern blot analysis and in situ mRNA hybridization (ISH) and protein production using enzyme-linked immunosorbent assay and immunohistochemistry in 8 human gastric carcinoma cell lines and 39 gastric carcinomas and corresponding normal mucosa (34 surgical specimens and 5 biopsy specimens). Of the 8 human gastric carcinoma cell lines, 6 expressed 1.8-kb IL-8 mRNA and secreted various levels of IL-8 protein. The expression of IL-8 by TMK-1 cells was induced by exposure to IL-1 alpha, epidermal growth factor, and transforming growth factor-alpha, shown previously to be autocrine growth stimulators for human gastric carcinoma cells. In tumor tissues, most of the tumors (28 of 34 surgical specimens and 4 of 5 biopsy specimens) expressed IL-8 at higher levels than the corresponding normal mucosa. ISH and immunohistochemical analyses revealed that IL-8 mRNA and protein were localized in the cytoplasm of tumor cells. The number of blood vessels in the gastric carcinomas was determined by using antibodies against CD34. The level of IL-8 mRNA in the neoplasms strongly correlated with vascularization (Spearman correlation, r = 0.812; P = 0.001). The data suggest that IL-8 produced by tumor cells may regulate neovascularization and, hence, the growth and spread of human gastric carcinoma.     

原位检测新基因SNC66在胃癌组织与正常胃粘膜中的表达

目的: 观察新基因SNC66在胃癌组织中的表达水平,分析SNC66表达水平与胃癌发生的相关性。 方法:以β-actin为对照,对20例胃癌组织及其配对正常胃粘膜的石蜡切片,进行原位cRNA/mRNA分子杂交和原位RT-PCR扩增。 结果:两种检测方法表明,SNC66在正常胃粘膜组织中都呈强阳性表达。而在胃癌组织中,原位分子杂交检测表明,20例标本SNC66不表达和弱阳性表达各8例,强阳性表达4例。原位RT-PCR检测表明,当循环数为10时,此20例胃癌标本中不表达、弱阳性和呈强阳性表达数分别为6,9和5;而当循环数为25时,则4例不表达,16例呈强阳性表达。 结论:SNC66基因在胃癌组织中存在明显的表达降低和表达缺陷。SNC66可以作为一个胃癌负相关基因加以进一步研究。     

Overexpression of tissue inhibitors of metalloproteinase-1 and -2 in the stroma of gastric cancer.

The fundamental event of cancer invasion and metastasis is the complicated interaction of cancer cells with host cells, in which event, a number of proteases and their inhibitors are involved. Matrix metalloproteinases are the potent proteases in degrading the basement membrane and extra cellular matrix and are inhibited by specific endogeneous inhibitors, tissue inhibitors of metalloproteinases-1(TIMP-1) and TIMP-2. The expression of mRNA for TIMP-1 and -2 was investigated by Northern blot analysis in specimens taken from 27 patients with primary gastric adenocarcinoma; 25 samples from the primary site, six from the metastatic lymph nodes and two from the peritoneal fluids. The expression for TIMP-1 and -2 was compared in primary gastric cancer tissues, metastatic lymph nodes and normal gastric mucosae. TIMP-1 mRNA was overexpressed in 24 (96%) out of 25 primary cancer tissues compared with the paired normal mucosae, while TIMP-2 was in 10 (40%). In six specimens of metastatic lymph nodes, TIMP-1 and -2 were overexpressed in 6 (100%) and 4 (67%) specimens, respectively. Of two specimens prepared from the peritoneal fluids, all specimens overexpressed TIMP-1 compared with the those of primary cancer tissues, while one (50%) specimen overexpressed TIMP-2. Immunohistochemical staining was done to investigate the localization of TIMP-1 and -2, demonstrating that the immunoreactivity for TIMP-1 and -2 was clearly detected in the cytoplasm of the stromal cells. These results suggest that both TIMP-1 and -2 are overexpressed by stromal cells in most of primary and some metastatic gastric cancer tissues and that TIMP-1 and TIMP-2, produced by stromal cells, may play an important role in inhibiting the proteolytic activity of matrix metalloproteinases originated from cancer cells, in gastric cancer.     

人类胃组织复合表达两种不同剪接的胆囊收缩素-B/胃泌素受体

为了探讨正常和异位剪接的胆囊收缩素 - B/胃泌素受体基因是否在人类胃上皮细胞中表达及其表达量与胃癌分化和转移的关系。检测了 30例胃癌和癌周正常胃组织上皮细胞、10例胃炎和 2例胃正常组织上皮细胞及胃癌细胞株 SGC- 790 1中这两种受体基因的表达水平。结果显示 ,正常、炎症和恶性胃上皮细胞同时表达正常和异位剪接的胆囊收缩素 - B/胃泌素受体 ,两种受体的表达量与胃癌的分化与转移无关。推测异位剪接的胆囊收缩素 -B/胃泌素受体在胃上皮细胞中以目前尚不清楚的功能发挥作用     

Serologic detection of infection with cagA+ Helicobacter pylori strains.

Approximately 60% of Helicobacter pylori isolates possess the cagA gene and express its 120- to 140-kDa product (CagA). In this study, the cagA gene was detected in H. pylori isolates from 26 (81.3%) of 32 patients with duodenal ulcers (DU), 17 (68.0%) of 25 patients with gastric ulcers, and 23 (59.0%) of 39 patients with nonulcer dyspepsia (NUD). By Western blotting (immunoblotting) with antiserum to CagA, in vitro CagA expression was demonstrated for 95.5% of cagA+ strains compared with 0% of strains lacking cagA. Sera from patients infected with cagA+ strains (n = 66) reacted with recombinant CagA in an enzyme-linked immunosorbent assay to a significantly greater extent than either sera from patients infected with strains lacking cagA (n = 30) or sera from uninfected persons (n = 25) (P < 0.001). A strain lacking cagA was isolated from eight patients who had serum immunoglobulin G antibodies to CagA, which suggests that these patients were infected with multiple strains. Serum immunoglobulin G antibodies to CagA were present in 87.5, 76.0, and 56.4% of patients with DU, gastric ulcers, and NUD, respectively (odds ratio, 5.41; 95% confidence interval, 1.44 to 24.72; P = 0.004 [DU versus NUD]). These data demonstrate an association between infection with cagA+ H. pylori and the presence of duodenal ulceration and indicate that serologic testing is a sensitive method for detecting infection with cagA+ strains.     

Induction of interleukin-8 secretion from gastric epithelial cells by a cagA negative isogenic mutant of Helicobacter pylori.

The ability of Helicobacter pylori strains to induce interleukin-8 (IL-8) gene expression and protein secretion from gastric epithelial cell lines in vitro is variable. This cellular response is associated with bacterial expression of the CagA protein present in type I H pylori strains. To determine the role of CagA in this host cell response, an isogenic cagA negative mutant, N6.XA3, was constructed. The cagA negative isogenic mutant and the wild-type parental cagA positive strain, N6, were cocultured with AGS, ST-42 and KATO-3 gastric epithelial cell lines and secreted interleukin-8 assayed by enzyme linked immunosorbent assay. In all three cell lines there was no significant difference in the IL-8 secretion induced by the cagA negative isogenic mutant, N6.XA3, and the wild-type parent strain, N6. These studies show that CagA is not the inducer of IL-8 secretion from gastric epithelial cells. As all wild-type CagA positive strains studied to date induce IL-8, the bacterial factor(s) inducing this inflammatory response is closely associated with the expression of CagA.