胃癌死亡相关蛋白激酶基因甲基化对其信使核糖核酸和蛋白表达的影响

目的 研究胃癌组织死亡相关蛋白激酶(DAPK)基因启动子区甲基化对原发性胃癌组织中DAPK mRNA及蛋白表达的影响.方法 采用逆转录(RT)-PCR法检测62例原发性胃癌及癌旁组织DAPK mRNA表达,甲基化特异性PCR(MSP)法检测DAPK启动子区CpG岛甲基化状态,对其中34例胃癌组织甲基化阳性者的DAPK蛋白表达进行Western印迹法检测.结果 胃癌组织中DAPKmRNA和蛋白表达水平明显低于癌旁组织,分别为0.2863±0.2027比0.5736±0.1968、0.2616±0.0913比0.6529±0.1808,差异均有统计学意义(P值均＜0.01).DAPK在胃癌组织和癌旁组织中的甲基化频率分别为54.8%和17.7%,差异有统计学意义(P＜0.01).在胃癌组织中,甲基化组DAPKmRNA表达明显低于非甲基化组(0.1399±0.0835比0.4640±0.1569,P＜0.01).DAPK基因甲基化与胃癌TNM分期显著相关(P=0.04).结论 原发性胃癌组织DAPK mRNA和蛋白表达缺失或低下与其启动子甲基化程度增高显著相关.     

死亡相关蛋白激酶在肝细胞癌中的表达及其临床意义

目的研究死亡相关蛋白激酶(death associated protein kinase,DAPK)在原发性肝细胞癌(HCC)组织中的表达及其与HCC临床病理特征的关系。方法应用免疫组织化学SP法检测DAPK在50例HCC组织及其癌旁组织和5例正常肝脏组织中的表达。结果 DAPK在HCC组织中的阳性率为36%,明显低于癌旁组织62%及正常肝组织100%(P<0.05);DAPK低表达与HCC组织分化程度、淋巴结转移有关(P<0.05)。结论 DAPK表达下调在HCC的发生发展中可能起重要作用。     

死亡相关蛋白激酶基因高甲基化在胃癌形成过程中作用的初步研究

背景：DNA启动子区甲基化可导致肿瘤抑制基因表达沉默，在胃癌的发生、发展中发挥重要作用。目的：观察正常胃黏膜、慢性萎缩性胃炎伴肠化生、异型增生和早期胃癌组织中死亡相关蛋白激酶（DAPK）基因启动子区甲基化状态，探讨其与胃癌发生、发展的关系。方法：以甲基化特异性聚合酶链反应（MSP）检测20例正常胃黏膜、14例慢性萎缩性胃炎伴肠化生、27例异型增生和16例早期胃癌组织中DAPK基因启动子区甲基化状态．并分析其与患者临床病理特征的关系。结果：早期胃癌组织中DAPK基因启动子区甲基化率显著高于正常胃黏膜、慢性萎缩性胃炎伴肠化生和异型增生组织（43.8％对0％、7．1％和11．1％，P〈0．05），而后三者之间DAPK基因启动子区甲基化率无明显差异。DAPK基因启动子区甲基化与患者性别、年龄和病变部位均无关，与幽门螺杆菌（H.pylori）感染和血清癌胚抗原（CEA）水平显著相关（P〈0．05）。结论：DAPK基因启动子区高甲基化是胃癌发生的早期分子事件，在由慢性萎缩性胃炎伴肠化生和异型增生进展至早期胃癌的过程中起重要作用。     

抑癌基因PTEN在胃癌中的表达及其临床意义

应用免疫组织化学SP法检测68例胃癌组织及33例癌旁正常组织中张力蛋白同源物基因（PTEN）蛋白的表达水平。结果癌旁正常组织中第10号染色体缺失的磷酸酶和PTEN蛋白阳性表达率为100％，胃癌中PTEN表达阳性率为61．8％；PTEN表达与胃癌的分化程度高低、浸润深度、是否有淋巴结转移和临床分期关系密切（P〈0．01，〈0．05），与患者年龄、肿瘤部位无关（P〉0．05）。PTEN异常表达可为胃癌诊断和判断其生物学特性提供可靠的指标。     

抑癌基因P15在胃癌中的表达及其临床意义

采用ＳＰ免疫组化法测定了８０例胃癌及２６例胃良性病变患者胃组织石蜡标本的Ｐ＾１５基因蛋白表达情况,结合临床病理指标（肿瘤大小、生长方式、组织分化、浸润深度、转移及ＰＴＮＭ分期）进行分析。结果在胃癌组织中Ｐ＾１５阳性表达率为４３．８％,胃良性病变为６９．２％;Ｐ＾１５表达与肿瘤大小、生长方式、组织分化、浸润深度,转移及ＰＴＮＭ分期无关。认为Ｐ＾１５在胃癌发生中起重要调控作用,与临床病理因素无关。     

ras相关结构域家族1A基因在胃癌组织中表达的意义

目的研究ras相关结构域家族(RASSF)1A基因在原发性胃癌组织中的表达及其与临床病理特征的关系,探讨在胃癌发生发展中的可能机制。方法收集54例原发性胃癌及癌旁正常组织,应用逆转录(RT)-PCR检测RASSF1A mRNA表达,Western印迹及免疫组化法检测RASSF1A蛋白表达。结果RT-PCR及Western印迹显示RASSF1A mRNA和蛋白表达水平,在胃癌组织中明显低于癌旁正常组织(A值分别为0.2589±0.2407比0.5448±0.2971;0.1874±0.0737比0.6654±0.2201, P值均<0.01)。免疫组化分析显示,RASSF1A在所有癌旁正常组织中表达均阳性,积分范围为1～12,均值7.94±3.75;而在胃癌组织中表达则明显减弱或缺失,积分范围为0～6,均值1.07±1.61。RASSF1A在癌旁组织中的表达水平显著高于胃癌组织(P<0.01)。此外,RASSF1A mRNA及蛋白表达水平与胃癌分期显著相关。结论在原发性胃癌组织中,RASSF1A mRNA和蛋白表达明显缺失或低下,且与胃癌临床分期显著相关。     

细胞凋亡调控基因Bcl—XL,Bcl—Xs在胃癌组织中的表达及临床意义

细胞凋亡调控基因与肿瘤发生的关系已成为当前肿瘤研究的热点。为此,我们应用免疫组化技术对６０例胃癌、癌旁组织及６１例癌前病变胃镜活检组织进行了细胞凋亡相关调控基因Ｂｃｌ－ＸＬ、Ｂｃｌ－ＸＳ蛋白的检测,结合临床病理学指标,探讨它们在胃癌、癌旁组织及癌前病变中的表达及临床意义。１．材料与方法１．１　一般资料　６０例胃癌及癌旁（距肿瘤边缘２ｃｍ）组织,４０例萎缩性胃炎伴肠上皮化生,２１例萎缩性胃炎伴不典型增生组织,３０例浅表性胃炎（对照）组织均为我院胃镜室１９９５年５月～１９９８年１月胃镜活检标本。所有…     

蛋白激酶B和15-脂氧合酶-1在胃癌中的表达及其临床意义

目的检测蛋白激酶B（Akt）和15-脂氧合酶-1（15-lipoxygenase-1,15-LOX-1）在胃癌及癌旁正常组织中的表达,以探讨二者在胃癌形成中的临床意义。方法采用RT-PCR法检测新鲜胃癌及癌旁正常组织标本中Akt-1、Akt-2、Akt-3和15-LOX-1 mRNA的表达;Western blot印迹法检测磷酸化Akt（p-Akt）和15-LOX-1蛋白的表达。结果Akt-1、Akt-2、Akt-3 mRNA在胃癌及癌旁正常组织中的表达水平差异均无统计学意义（P〉0.05）。p-Akt蛋白在胃癌组织中的表达显著高于癌旁正常组织（P〈0.05）。15-LOX-1蛋白及mRNA在胃癌组织的表达均低于癌旁正常组织（P〈0.05）。p-Akt蛋白表达与胃癌的肿瘤大小、淋巴结转移和TNM分期呈正相关（P〈0.05）。相关性分析表明：胃癌组织中15-LOX-1蛋白表达和p-Akt水平间无明显相关性（r=-0.252,P〉0.05）。结论Akt在胃癌的发生发展中具有促进作用。15-LOX-1蛋白可能对胃癌的发生发展有一定抑制作用,但与Akt无直接关系。     

Bmi-1在胃癌中的表达及其意义

目的探讨Bmi-1在胃癌和癌旁组织中的表达水平及其与临床病理特征的关系，初步评价Bmi-1在胃癌发生、发展中的作用及其意义。方法收集20例胃癌患者的癌组织和癌旁组织，采用Western印迹法和RT-PCR检测Bmi-1表达。同时收集2002-2004年146例有3年以上完整随访资料的胃癌术后患者，采用免疫组化法对手术标本进行染色，检测Bmi-1蛋白表达。结果Western印迹和RT-PCR结果均显示，胃癌组织中Bmi-1表达水平明显高于癌旁组织。免疫组化分析也表明，Bmi-1蛋白在胃癌中阳性表达率为67．8％（99／146）。Bmi-1的表达与胃癌大小、临床分期、淋巴结转移和浸润深度密切相关（P〈0．05），而与患者性别、年龄、肿瘤分化程度等无关（P〉0．05）。结论Bmi-1在胃癌组织的表达状态与胃癌生长和浸润转移关系密切，可作为反映胃癌生物学行为的有效指标。     

胃癌中HER-2/neu基因扩增和蛋白表达的多中心研究

目的 观察中国人胃癌HER-2／neu基因扩增和蛋白表达状况，探讨两者间关系及与患者临床病理参数间的关系。方法 前瞻性研究中挑选胃腺癌蜡块270例，回顾性研究中挑选蜡块277例，分别用显色原位杂交（CISH）和免疫组化法（IHC）检测HER-2／neu基因扩增和蛋白表达状况。结果 胃癌前瞻性病例中HER-2／neu基因扩增率为14．8％，蛋白过表达率为11．9％。肠型胃癌HER-2／neu基因扩增率和蛋白过表达率分别为25．4％和18．8％，明显高于弥漫型胃癌的4．7％和5．5％（P值均〈0．01）。高、中度分化胃癌HER-2／neu基因扩增率和蛋白过表达率均明显高于低分化胃癌（P值均〈0．01）。回顾性、前瞻性研究及汇总后的总结果中HER-2／neu蛋白CISH和IHC检测结果符合率分别为77．0％、89．2％、83．2％，两者显著相关（P值均〈0．01）。结论 HER-2／neu基因扩增、蛋白过表达可能与中国人胃癌发生有关，基因扩增可能是其蛋白过表达的主要分子机制，HER-2／neu基因扩增和蛋白过表达与肠型胃癌和高中度分化胃癌相关。     

Clinicopathological significance of FHIT protein expression in gastric adenocarcinoma patients

AIM: To investigate the expression of fragile histidine triad(FHIT) protein, and the possible relationship between FHIT expression and clinicopathological indices in gastric carcinoma. METHODS: FHIT protein expression was examined in 76 cases of gastric carcinoma, 58 cases of intraepithelial neoplasia, and 76 cases of corresponding normal mucosae by immunohistochemical method to analyze its relationship to histological grade, clinical stage, metastatic status and prognosis. RESULTS: The FHIT protein expression was positive in 28/76(36.8%) cases of adenocarcinoma tissue, 22/58(37.9%) cases of adjacent dysplastic tissue and 76/76(100%) cases of distal normal gastric mucosa. There was a significant difference in the expression of FHIT protein between cancer or adjacent intraepithelial neoplasia and normal gastric mucosa(P=0.000). FHIT protein expression was found in 64.3%(18/28) of grades Ⅰ and Ⅱ cancers, and 20.8%(10/48) of grade Ⅲ cancers(P=0.000), in 56.3%(18/32) of stages Ⅰ and Ⅱ cancers and 22.7%(10/44) of stages Ⅲ and Ⅳ cancers(P=0.004), and in 63.6%(14/22) of cancers without metastasis but only 25.9%(14/54) of those with metastasis(P=0.003). The significant difference in the expression of FHIT was found between histological grade, clinical stage and metastatic status of cancer. Follow-up data showed that there was a significant difference in median survival time between cancer patients with expression of FHIT(71 mo) and those without (33 mo, log rank=20.78, P=0.000). CONCLUSION: FHIT protein is an important tumor suppressor protein. Loss of FHIT protein expression may be associated with carcinogenesis, invasion, metastasis and prognosis of gastric adenocarcinoma.     

用基因表达谱芯片研究人正常胃和胃癌中与发育相关基因的价值

用8464条人cDNA制成表达谱芯片,利用胃和胃癌组织的mRNA,通过逆转录方法将Cy3和Cy5两种荧光分别标记到两组cDNA上,利用这种cDNA探针与表达谱芯片进行杂交后扫描,通过计算机分析判定基因是否在上述组织中存在差异表达,结果示在197条与生长发育相关的基因中,存在差异表达的共10条,上调的8条（0．095％）,下调的2条（0．024％）,应用这种方法识别出的基因对胃癌的诊断和治疗具有重要的潜在价值。     

Gene expression profile differences in gastric cancer, pencancerous epithelium and normal gastric mucosa by gene chip

AIM: To study the difference of gene expression in gastric cancer (T), pericancerous epithelium (P) and normal tissue of gastric mucosa (C), and to screen an associated novel gene in early gastric carcinogenesis by oligonudeotide microarray. METHODS: U133A (Affymetrix, Santa Clara, CA) gene chip was used to detect the gene expression profile difference in T, P and C, respectively. Bioinformatics was used to analyze the detected results. RESULTS: When gastric cancer was compared with normal gastric mucosa, 766 genes were found, with a difference of more than four times in expression levels. Of the 766 genes, 530 were up-regulated (Signal Log Ratio [SLR]>2), and 236 were down-regulated (SLR<-2). When pericancerous epithelium was compared with normal gastric mucosa, 64 genes were found, with a difference of more than four times in expression levels. Of the 64 genes, 50 were up-regulated (SLR>2), and 14 were down-regulated (SLR<-2). Compared with normal gastric mucosa, a total of 143 genes with a difference in expression levels (more than four times, either in cancer or in pericancerous epithelium) were found in gastric cancer (T) and pericancerous epithelium (P). Of the 143 genes, 108 were up-regulated (SLR>2), and 35 were down-regulated (SLR<-2). CONCLUSION: To apply a gene chip could find 143 genes associated with the genes of gastric cancer in pericancerous epithelium, although there were no pathological changes in the tissue slices. More interesting, six genes of pericancerous epithelium were up-regulated in comparison with genes of gastric cancer and three genes were down-regulated in comparison with genes of gastric cancer. It is suggested that these genes may be related to the carcinogenesis and development of early gastric cancer.     

Gene expression profile differences in gastric cancer, pericancerous epithelium and normal gastric mucosa by gene chip

AIM: To study the difference of gene expression in gastric cancer (T), pericancerous epithelium (P) and normal tissue of gastric mucosa (C), and to screen an associated novel gene in early gastric carcinogenesis by oligonucleotide microarray. METHODS: U133A (Affymetrix, Santa Clara, CA) gene chip was used to detect the gene expression profile difference in T, P and C, respectively. Bioinformatics was used to analyze the detected results. RESULTS: When gastric cancer was compared with normal gastric mucosa, 766 genes were found, with a difference of more than four times in expression levels. Of the 766 genes, 530 were up-regulated (Signal Log Ratio (SLR) >2), and 236 were down-regulated (SLR<-2). When pericancerous epithelium was compared with normal gastric mucosa, 64 genes were found, with a difference of more than four times in expression levels. Of the 64 genes, 50 were up-regulated (SLR>2), and 14 were down-regulated (SLR<-2). Compared with normal gastric mucosa, a total of 143 genes with a difference in expression levels (more than four times, either in cancer or in pericancerous epithelium) were found in gastric cancer (T) and pericancerous epithelium (P). Of the 143 genes, 108 were up-regulated (SLR>2), and 35 were down-regulated (SLR<-2). CONCLUSION: To apply a gene chip could find 143 genes associated with the genes of gastric cancer in pericancerous epithelium, although there were no pathological changes in the tissue slices. More interesting, six genes of pericancerous epithelium were up-regulated in comparison with genes of gastric cancer and three genes were down-regulated in comparison with genes of gastric cancer. It is suggested that these genes may be related to the carcinogenesis and development of early gastric cancer.     

Clinical implication of 14-3-3 epsilon expression in gastric cancer

AIM: To evaluate for the first time the protein and mRNA expression of 14-3-3ε in gastric carcinogenesis.METHODS: 14-3-3ε protein expression was determined by western blotting, and mRNA expression was examined by real-time quantitative RT-PCR in gastric tumors and their matched non-neoplastic gastric tissue samples.RESULTS: Authors observed a significant reduction of 14-3-3ε protein expression in gastric cancer (GC) samples compared to their matched non-neoplastic tissue. Reduced levels of 14-3-3ε were also associated with diffuse-type GC and early-onset of this pathology. Our data suggest that reduced 14-3-3ε may have a role in gastric carcinogenesis process.CONCLUSION: Our results reveal that the reduced 14-3-3ε expression in GC and investigation of 14-3-3ε interaction partners may help to elucidate the carcinogenesis process.     

DNA甲基化和胃液固有荧光光谱联合检测在胃癌诊断中的意义

目的探索在胃癌患者胃液、外周血清中检测基因甲基化的可行性，并结合胃液稀释固有荧光光谱评价二者在胃癌诊断中的作用。方法采用甲基化特异性PCR方法，检测50例胃癌患者的原发肿瘤组织、外周血清和胃液脱落细胞的死亡相关蛋白（DAP）激酶、p16基因启动子区域甲基化状态，并以胃良性溃疡和慢性浅表性胃炎各20例、慢性萎缩性胃炎30例作为对照，同时检测胃癌患者和对照者的胃液稀释固有荧光光谱。结果50例胃癌患者的肿瘤组织、外周血清和胃液脱落细胞中p16和DAP激酶基因甲基化阳性率分别为74．4％和68．1％、52．0％和58．0％、58．6％和76．0％，20例慢性浅表性胃炎患者中均未检测到基因异常甲基化；20例胃溃疡患者溃疡周边组织、胃液脱落细胞中p16和DAP激酶甲基化阳性率为10．0％和20．0％、5．0％和15．0％，在外周血清中未检测到异常甲基化；30例慢性萎缩性胃炎患者胃黏膜组织、外周血清、胃液脱落细胞p16和DAP激酶基因甲基化阳性率分别为10．0％和23．3％、3．3％和3．3％、3．3％和20．0％。胃癌患者胃液固有荧光光谱强度较对照者明显增强（P〈0．05）；以P1FI≥111．8为分界点分析胃液固有荧光光谱诊断胃癌的敏感性和特异性分别为76．1％和78．6％。p16和DAP激酶基因甲基化和胃液固有荧光光谱结合后诊断胃癌的敏感性提高到95．6％和97．8％。结论胃癌患者外周血清及胃液脱落细胞中可检测到与原发肿瘤组织一致的基因异常甲基化，胃液固有荧光光谱和DNA甲基化联合对检测胃癌有良好的临床应用价值。     

人巨噬细胞金属弹性蛋白酶在胃癌细胞株及胃癌组织中的表达及意义

目的测定人巨噬细胞金属弹性蛋白酶（HME）在胃癌细胞及胃癌组织中的表达,评价其在胃癌进展中的作用.方法2003年4月至8月胃肠外科和肿瘤外科胃癌住院手术病人共58例,所有病人均取肿瘤组织、癌旁组织、远处正常组织标本,采用R-PCR、实时荧光定量PCR测定胃癌细胞株（MGC-803、SGC-7901、AGS）及胃癌组织中HME mRNA水平,采用Western印迹和免疫组化法测定HME蛋白水平.结果3株胃癌细胞株中均有HME mRNA和蛋白表达;胃癌组织中的HMEmRNA和蛋白表达高于远处正常组织,差异有统计学意义（P＜0.05）;而胃癌组织与癌旁组织中的HME mRNA和蛋白表达差异无统计学意义（P＞0.05）.结论胃癌患者中HME mRNA和蛋白呈一定比例的阳性表达,胃癌组织中最高,其次为癌旁组织和远处正常组织,说明HME是一项有潜在价值的肿瘤相关标志物.