人胎盘型谷胱甘肽S转移酶在胃癌组织中的表达与基因甲基化关系？…

目的：研究胃癌组织中ＧＳＴ－π的表达与其基因甲基化状态的关系。方法：应用Ｓ－Ｐ法检测１１６例胃癌和５３例胃癌前病变的ＧＳＴ－π表达和限制性内切酶及ＰＣＲ、Ｓｏｕｔｈｅｒｎ印迹方法检测１４例胃癌及其相应正常胃粘膜ＧＳＴ－πＤＮＡ５端调我ＣＣＧＧ特定位点的甲基化水平。结果：ＧＳＴ－π阳性率,正常胃癌膜１０％,胃癌７７％,肠上皮化生７６％,不典型增生８９％。ＧＳＴ－π在胃癌及癌前病变中的表达较正常胃粘膜     

胃癌中谷胱甘肽S—转移酶的表达

研究胃癌组织中谷胱甘肽Ｓ－转移酶（ｇｌｕｔａｔｈｉｏｎｅＳ-ｔｒａｎｓｆｅｒａｓｅｓ,ＧＳＴｓ）的表达。方法应用免疫组织化学方法研究ＧＳＴ－π在９８例胃癌标本中的表达。结果在９８例胃癌标本中,ＧＳＴ－π阳性率在不同的病理组织类型中为２５．０％～８８．９％之间。ＧＳＴ－π在低分化胃癌高于高分化胃癌。但与患者的年龄、性别、肿瘤的大小、部位、淋巴结转移情况等无明显关系。结论ＧＳＴ－π对胃癌的诊断、预后及治疗有重要价值。     

人胃癌组织中C—myc癌基因甲基化及其表达的研究

目的：研究胃癌及正常胃粘膜ｃ－ｍｙｃ基因外显子ⅢＣＣＧＧ位点的甲基化状态及其ｍＲＮＡ表达及ＳＡＢＣ法检测胃癌４５例、胃溃疡１４例、不典型增生１０例ｐ６７表达。结果：胃癌ｃ－ｍｙｃ基因外显子ⅢＣＣＧＧ位点比其相应正常胃粘膜呈高度去甲基化，ｍＲＮＡ表达升高（ｐ〈０．００１），ＤＮＡ去甲基化与ｍＲＮＡ表达存在相关性（ｐ〈０．０２５），ｐ６７在胃癌、不典型增生、胃溃疡表达率分别为８４．４４％（３８／４５）     

PTEN抑癌基因在人胃癌组织中的表达及意义

目的探讨抑癌基因PTEN在胃癌组织中的表达水平及与其生物学行为的关系。方法应用免疫组织化学EnVision法检测116例胃癌组织,35例癌旁异型组织,56例癌旁正常黏膜组织中的PTEN蛋白表达情况。结果胃癌组织中PTEN阳性率为53.4 %(62/116),显著低于异型增生、癌旁正常黏膜组织中的阳性率71.4 %(25/35)、100 %(56/56)(P<0.001)。在各组织学类型中,高中分化腺癌PTEN蛋白阳性表达率最高(72.7 %,32/44),显著高于印戒细胞癌(29.2 %,7/24),P<0.001。PTEN蛋白的表达与淋巴结转移、侵袭程度密切相关(P<0.05)。结论PTEN基因的异常改变与胃癌的发生发展密切相关,可能是胃癌进一步恶化的标志之一,可作为胃癌生物学行为的参考指标,为胃癌的基因治疗提供参考依据。     

GST-π和P-gp蛋白在胃癌中的表达

目的　研究胃癌组织中GST π和P gp糖蛋白表达的意义。 方法　应用SP免疫组织化学的方法对 98例胃癌进行检测。结果　在 98例胃癌中 ,75 .5 %的GST π表达阳性 ,5 7.1%的P 糖蛋白表达阳性。GST π和P 糖蛋白与胃癌的分化程度有关 (P <0 .0 5 )。而与患者的年龄、性别、肿瘤的大小、部位、浸润程度和淋巴结转移情况无关。结论　联合检测GST π和P 糖蛋白对胃癌的诊断、治疗和预后有重要意义。     

肺癌组织酸性谷胱甘肽S—转移酶基因测定及其临床意义

研究运用ＲＴ－ＰＣＲ方法对５１例手术切除肺癌组织和相应的癌周正常肺组织进行ＧＳＴ－πｍＲＮＡ水平检测。结果表明：ＧＳＴ－πｍＲＮＡ在正常肺组织和肺癌组织中表达阳性率分别为１９．６％（１０／５１）和５１．０％（２６／５１），两者之间具有明显差别。ＧＳＴ－πｍＲＮＡ表达与肿瘤病理类型、组织分化、ＴＮＭ分期等均无明显的相关性（Ｐ＞０．０５）。上述结果表明肺组织细胞在恶变过程中ＧＳＴ－πｍＲＮＡ水平表达明显增加，提示ＧＳＴ－π基因是肺癌肿瘤标志之一，同时在肺癌先天性耐药机制中占有重要的作用     

GSTP1基因在前列腺癌组织中的表达及甲基化状态研究

目的：研究谷胱苷肽S转移酶P1（glutathione S transferase pi,GSTP1）在前列腺癌的表达情况并分析其甲基化状态.方法：采用免疫组织化学方法检测GSTP1在50例前列腺癌石蜡标本中的表达情况,并用甲基化特异性聚合酶链反应（methylation-specific polymerase chain reaction, MSP）方法分析GSTP1基因启动子区5＇端CpG岛非甲基化和甲基化的状态.结果： GSTP1在前列腺癌表达的阳性率为18.0%,明显低于癌旁正常前列腺腺体的阳性率（76.0%）,P=0.000.GSTP1基因启动子区非甲基化发生率为86%（43/50）,50例前列腺癌均未扩增出甲基化特异性产物.结论： GSTP1阴性表达有望成为前列腺癌诊断的一项辅助性新指标.国内前列腺癌GSTP1基因的高频率表达缺失可能与GSTP1基因启动子区甲基化无关,因本组病例数较少,还需进一步积累资料证实.     

Pi类谷胱甘肽S—转移酶与消化道肿瘤

近年来研究发现谷胱甘肽S－转移酶（GST）与癌的发生,发展及抗药性的产生有着密切的关系,其中同工酶Pi类谷胱甘肽S－转移酶（GST－π）在消化道肿瘤中有一定表达,在其相应的癌、癌前病变组织及血中含量增多,可作为消化道肿瘤的一个标志酶,有助于消化道肿瘤的早期诊断的疗效观察。     

Expression of Sulfated Carbohydrate Chains Detected by Monoclonal Antibody 91.9H in Human Gastric Cancer Tissues

Expression of sulfated carbohydrate chains in intestinal metaplasia (IM) and gastric cancer tissues was immunohistochemically evaluated by using a monoclonal antibody (MAb) 91.9H. While normal gastric tissues were negative for MAb 91.9H, IM and cancer tissues were positively stained with high frequency. The incidence was 67% for IM with chronic gastritis (n = 12), 95% for IM with gastric cancer (n=21), 77% for well and moderately differentiated adenocarcinomas (n = 13), 48% for poorly differentiated adenocarcinoma (n = 23), 89% for signet-ring cell carcinoma (n = 9) and 100% for mucinous adenocarcinoma (n = 7). When poorly differentiated adenocarcinoma cases were divided into two groups, solid (n = 13) and non-solid types (n = 10), the incidences were 8% and 100%, respectively. These data suggest that MAb 91.9H could be of practical use as a new marker for IM and gastric cancer, and may be valuable for subgrouping poorly differentiated adenocarcinomas. Analyses of the core proteins for 91.9H epitope were then carried out. Comparison of immunostaining of ten poorly differentiated adenocarcinoma cases by MAb MUSE11 against MUC1 gene product with that by MAb 91.9H suggested that 91.9H epitope is not expressed on MUC1. Northern blot analysis of 10 pairs of gastric cancer and adjacent normal tissues with a MUC2 cDNA probe showed that the expression level of MUC2 mRNA was below the limit of detection. Thus, 91.9H epitope may be expressed on other proteins than MUC1 or MUC2 core proteins in gastric cancer.     

Methylation and Expression of Human Pepsinogen Genes in Normal Tissues and Their Alteration in Stomach Cancer

In normal human tissues, pepsinogen A mRNA was expressed only in the fundic mucosa of the stomach, whereas pepsinogen C mRNA was expressed in all regions of the stomach mucosa and also in the proximal duodenal mucosa. The distributions of these mRNAs were consistent with those of pepsinogens A and C in the gastroduodenal mucosa. Methylation analysis of DNAs from normal tissues with methylation-sensitive restriction enzymes, Hpa II and Hha I, revealed that pepsinogen A and C genes are hypomethylated in tissues producing pepsinogens A and C, suggesting a role of DNA methylation in the regulation of the differential expression of the genes for the two human pepsinogens during normal differentiation. In stomach cancer tissues and cancer cell lines, the expressions of the pepsinogen genes were decreased or lost, in good accordance with their pepsinogen productions. No gross structural changes of the pepsinogen genes were observed in these cancers, but the methylation patterns of the pepsinogen genes were found to be altered in different ways in different cancers. The functional significance of the altered methylation is unknown; however, these results suggest that considerable heterogeneity of the methylation patterns occurs in human stomach cancers.     

人胃癌组织KiSS—1基因表达水平的检测

为了研究胃癌组织中ＫｉＳＳ１的表达水平及其与胃癌临床病理特征的相关性,探讨ＫｉＳＳ１表达用于预测胃癌淋巴结转移的可能性。采用逆转录聚合酶链式扩增反应检测了３０例胃癌组织ＫｉＳＳ１的表达水平。结果３０例胃癌组织中９例（３０．０％）有阳性ＫｉＳＳ１表达,ＫｉＳＳ１表达与胃癌的淋巴结转移、肿瘤的浸润深度呈负相关,与胃癌的Ｂｏｒｒｍａｎｎ分型、肿瘤的分化程度无关。ＫｉＳＳ１表达可能在胃癌的转移过程中起一定作用。     

胃癌相关的人源单链抗体基因文库的构建

目的　构建胃癌相关的人源单链抗体基因文库。方法　应用DNA重组技术 ,从胃癌患者的转移淋巴结及其脾细胞mRNA中构建出组合单链抗体 (ScFv)cDNA文库。将cDNA文库克隆到噬菌粒载体 pCANTAB5 ,并转化大肠杆菌TG 1。随机对所得细菌克隆进行序列分析。结果　cDNA文库克隆到噬菌粒载体pCANTAB5后转化大肠杆菌TG 1,得到 2 .5× 10 7个氨苄青霉素菌落。ScFv序列分析结果证实VH 属于人源抗体VHⅢ亚群 ,VL 属于VκⅢ亚群。结论　胃癌相关的人源单链抗体基因文库的构建为进一步构建噬菌体单链抗体库奠定了基础     

Pepsinogens I and II in Gastric Cancer: An Immunohistochemical Study Using Monoclonal Antibodies

Monoclonal antibodies were used to examine the immunohistochemical expression of pepsinogens I and II in 31 early and 76 advanced gastric cancers. Of the 107 carcinomas studied, 19 contained pepsinogen II and only 3, found exclusively in pepsinogen II-positive cases, contained pepsinogen I. Gastric cancer produces pepsinogen II more frequently than pepsinogen I, and production of the latter is significantly associated with the former. Historically, there were 54 intestinal-type and 53 diffuse-type cancers. The former produced pepsinogen II more frequently than the latter. In the diffuse type, the four pepsinogen II-positive cases were found exclusively in females. Although the pepsinogen expression was independent of the macroscopic features in advanced gastric cancer, it was found that the protruded-type early gastric cancer produced pepsinogen II more frequently than the depressed type. Incidences of pepsinogen positivity were not different between early and advanced gastric cancers or between cancers with or without lymph node metastasis, suggesting that production of pepsinogen is independent of tumor growth.     

MG7免疫组化法早期诊断胃癌的意义

对273例胃粘膜活检标本行单克隆抗体(McAb)M_(G7)PAP法免疫组化染色,部分标本同时行免疫电镜及粘液组化染色。结果:76例胃癌中69例M_(G7)阳性(90.8％),96例良性病伴不典型增生和/或肠化中17例M_(G7)阳性(17.7％),良性病中Ⅱb型肠化的M_(G7)阳性率(53.3％)高于Ⅱa型肠化(15.4％)。免疫电镜显示M_(G7)阳性物质分布于癌细胞膜、胞浆及个别良性病细胞微绒毛上。1年内随访7例良性病M_(G7)阳性者,2例发展为高度疑癌。本文结果提示:对胃癌癌前病变行McAb M_(G7)免疫组化染色,有助于筛选高危人群。最终可能发现早期胃癌。     

抗乳腺癌单克隆抗体的制备及其应用

目的：制备抗人乳腺癌单克隆抗体（McAb），用于临床肿瘤的免疫病理诊断与分型研究，为药物导向诊断和治疗奠定基础。方法：以乳腺癌细胞系ZR75免疫Balb/c小鼠，取其脾细胞与小鼠骨髓瘤细胞SP2／0进行融合，经选择培养、筛选及克隆化，获得恒定分泌抗ZR76单克隆抗体的杂交瘤细胞系BM109。结果：单抗BM109的Ig亚类为IgG1，BM109抗原主要表达在靶细胞膜上，为分子量37KD的蛋白质抗原。在乳腺癌的免疫反应率为86．7％（26／30），与胃癌、食管癌、卵巢癌等6种肿瘤的免疫反应率为9．5％（4／42），而对乳腺纤维瘤、囊性增生症和周围正常乳腺组织未见阳性反应。结论：所获一株抗乳腺癌单克隆抗体，具有较好的特异性，可望进一步用于乳腺癌的诊断、治疗及其发生发展的研究。     

胃癌CD44v6表达及其临床意义的研究

探讨胃癌ＣＤ４４ｖ６的表达,评价其表达对胃癌进展程度和转移的诊断意义,以及对手术预后的影响。方法收集胃癌手术标本８５例,相应的癌转移淋巴结标本４７例,应用Ｓ－Ｐ免疫组化染色技术检测ＣＤ４４ｖ６蛋白在上述组织中的表达。结论：对胃癌组织进行ＣＤ４４ｖ６蛋白的检测,有助于预测胃癌进展程度和淋巴结转移的诊断,以及评估胃癌患者的预后。     

Expression of CD44 Variant Exons 8-10 in Gastric Cancer

The expression of CD44 variant containing variant exons 8-10 product (CD44v8-10) was studied by western blot analysis and immunohistochemistry in gastric cancers using a monoclonal antibody, 44-IV. On western blots, a single band of 130 kD was recognized in stomach cancer cell lines. CD44v8-10 expression, with reactivity localized in the cell membrane, was found in 65 (33.5%) of the 194 advanced gastric cancers. There was no correlation between CD44v8-10 immunoreactivity and serosal, lymphatic, or lymph node invasion. However, there was significant correlation with CD44v8-10 immunoreactivity and venous invasion. CD44v8-10-positive cancers were more frequently associated with hematogenous metastasis than those which were immunonegative. There was an inverse association between CD44v8-10 immunoreactivity and peritoneal dissemination, especially in diffuse type adenocarcinomas. These observations indicate that CD44v8-10 may play a role in the metastasis of gastric cancer.     

胃癌ER基因表达与CpG岛甲基化的意义

目的:研究雌激素受体(ER)基因表达及其CpG岛甲基化状况与胃癌生物学行为和预后的关系.方法:分别应用免疫组化SP法和限制性内切酶PCR法分析91例胃癌ER的表达和30例胃癌ER基因CpG岛的甲基化状况.结果:胃癌ER阳性率为38%(35/91),其中高分化腺癌10%(3/30),分别与低分化腺癌43%(13/30),未分化癌53%(8/15),粘液癌63%(5/8)及印戒细胞癌75%(6/8)比较,皆具有显著性差异(P＜0.05).BorrmannⅢ Ⅳ型56%(24/43)明显高于Ⅰ型22%(4/18)与Ⅱ型23%(7/30)(P＜0.05).淋巴结转移组为55%(28/51),未转移组23%(9/40),两组间有显著性差异(P＜0.05).正常胃粘膜ER基因CpG岛未见甲基化,而40%(12/30)胃癌呈甲基化(P＜0.05),且与其蛋白表达有负相关性(P＜0.05).结论:ER在胃癌中的表达与分化、侵袭、转移有关,并以分化较差的胃癌表达高,提示ER阳性胃癌生物学行为差而预后不佳.ER基因CpG岛甲基化可能是胃癌中ER表达缺失的分子机制.