胃癌组织中CTGF蛋白表达及其临床意义

目的探讨结缔组织生长因子（CTGF）蛋自在胃癌发生、发展中的作用。方法采用免疫组化法检测10份正常胃黏膜组织、50份胃癌组织及其癌旁组织中CTGF蛋白表达情况。结果胃癌组织中CTGF阳性表达高于癌旁组织及正常胃黏膜组织,P〈0．05;癌组织CTGF阳性表达率明显高于正常黏膜组织（P〈0．01）,CTGF的高表达与胃癌分化程度、淋巴结转移密切相关（P均〈0．05）;与肿瘤浸润的深度无明显关系。结论CTGF蛋白可作为胃癌前病变及胃癌早期诊断和预后判断的指标。     

MTAl蛋白在胃癌中的表达及临床意义

应用免疫组化SP法检测67例胃癌、31例癌旁非典型增生及20例正常胃黏膜组织的MTAl蛋白表达，分析其与胃癌发生发展、浸润转移的关系。结果显示，MTAl蛋白在正常胃黏膜、癌旁非典型增生和胃癌组织中的表达率分别为5．00％、22．58％和67．16％，组间比较均有统计学差异（P均〈0．05）；MTAl蛋白表达与胃癌浸润程度、分化程度及淋巴结转移密切相关（P均〈0．05）。提示MTAl参与胃癌的发生发展。     

Cx43和Fas在胃癌组织中的表达

目的研究间隙连接蛋白43（Cx43）和凋亡调节因子Fas在胃癌组织中的表达情况。方法应用免疫组化法检测70例胃癌患者手术切除的癌组织和癌旁正常组织中Cx43和Fas表达水平。结果Cx43在胃癌组织中的表达率为44．29％（31／70）,在癌旁正常组织中的表达率为92．86％（65／70）,P〈0．01;Fas在胃癌组织中的表达率为40．00％（28／70）,在癌旁正常组织中表达率为87．14％（61／70）,P〈0．01。Cx43表达水平与胃癌组织学分化程度、浸润深度、淋巴结转移有关（P〈0．05）,Fas表达水平与胃癌组织学分化程度、浸润深度、TMN分期、淋巴结转移有关（P〈0．05）。结论胃癌组织中Cx43和Fas表达水平的检测有助于判断胃癌的恶性程度。     

SIRT1在胃癌组织、细胞中的表达及意义

目的观察SIRTl在胃癌组织及胃癌细胞中的表达情况,探讨SIRTl同胃癌临床病理特征之间的关系。方法应用Real．timePCR方法检测SIRTlmRNA在21例新鲜胃癌及癌旁组织中的表达情况;应用Westernblot和Real．timePCR方法在蛋白和mRNA水平检测SIRT1在胃癌、胃黏膜上皮细胞中的表达;应用免疫组织化学染色法检测130例胃癌组织及癌旁正常组织中SIRT1蛋白的表达,分析SIRTl蛋白表达与胃癌临床病理特征的关系。结果SIRTImRNA在新鲜胃癌组织中较正常癌旁组织表达增高（P〈0．05）,SIRT1mRNA及蛋白在胃癌细胞中均比正常胃黏膜上皮细胞中表达增高（P均〈0．05）。免疫组化染色结果表明,SIRTl蛋白在胃癌组织中较正常癌旁组织表达增高（P〈0．05）。SIRT1蛋白表达水平同胃癌患者的年龄及肿瘤浸润深度、淋巴结转移、TNM分期、大小有关（P均〈0．05）,同生存时间呈负相关（r=-0．5023,P〈0．05）。结论SIRT1在胃癌组织和胃癌细胞中呈高表达,其在胃癌发生中充当癌基因的角色。     

胃癌组织中Ptc蛋白的表达变化及意义

目的观察胃癌组织中Ptc蛋白的的表达情况，并探讨其与胃癌分化和转移的关系。方法采用West—ern—Blot法检测15例胃癌患者肿瘤和癌旁组织中的Ptc蛋白。采用免疫组化SP法检测45例胃癌患者肿瘤组织标本中的Ptc蛋白，并分析其与胃癌临床病理参数的关系。结果胃癌组织中Ptc蛋白表达量为0．375±0．117，低于癌旁组织的0．697±0．158，P〈0．05。胃癌组织中Ptc的表达水平与肿瘤组织分化程度、临床分期和远处转移有相关性（P均〈0．05）。结论胃癌组织Ptc蛋白表达降低，其与胃癌分化与转移程度密切相关。     

胃癌组织中CHD1L表达及其临床意义

目的：探讨染色质解旋酶DNA结合蛋白1样基因（CHD1L）在胃癌发生发展中的作用。方法采用实时定量RT-PCR和Western blot法观察3株人胃癌细胞系（AGS、N87、HGC-27）及正常胃黏膜上皮细胞中CHD1L mRNA和蛋白表达的差异;采用免疫组化法检测71例人胃癌组织及其配对癌旁组织中CHD1L蛋白表达,分析CHD1L与胃癌临床病理特征的关系。结果与正常胃黏膜上皮细胞比较,3株胃癌细胞系CHD1L mRNA和蛋白表达水平均明显上调（P＜0．05）;71例胃癌组织中CHD1L蛋白表达阳性率明显高于癌旁组织（P＜0．05）,CHD1L蛋白阳性表达与胃癌TNM分期、分化程度及淋巴结转移显著相关（P＜0．05）,与患者性别、年龄及肿瘤大小无相关性（P＞0．05）。结论 CHD1L在胃癌的发生发展中起促进作用;CHD1L表达有助于判定胃癌恶性表型。     

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。     

蛋白激酶B在胃癌中的表达及其生物学意义

目的研究丝氨酸／苏氨酸蛋白激酶（Akt）在胃癌中的表达、活化情况及其与血管内皮生长因子C（VEGF—C）的关系，以初探Akt表达和活化的生物学意义及其可能的机制。方法采用RT—PCR法检测20例新鲜胃癌及癌旁正常组织标本中Akt1、Akt2和Akt3 mRNA的表达；Western印迹法检测Akt和磷酸化Akt（pAkt）蛋白的表达；免疫组化检测pAkt蛋白和VEGF-C在55例胃癌配对组织中的原位表达。结果20例胃癌及相应癌旁正常组织中Akt1、2、3mRNA的表达阳性率皆为100％，且三者在癌和正常组织中的表达水平差异无统计学意义（P〉0．05）。Western印迹显示Akt蛋白在胃癌组织中的表达水平为正常组织的2．7倍（P〈0．001），而pAkt蛋白的表达水平为癌旁正常组织的4．1倍（P％0．01）。免疫组化分析表明，pAkt表达于67．3％（37／55）的胃癌组织，且较高表达于组织分化较差（X^2=9．751，P％0．01）和TNM分期较晚（X^2=7．684，P〈0．05）者；VEGF—C在胃癌组织中的表达阳性率为61．8％（34／55），其表达与肿瘤的TNM分期（X^2=9．298，P〈0．01）及淋巴结转移（X^2=7．605，P〈0．05）密切相关，统计分析表明，pAkt与VEGF—C的表达呈显著性正相关（Pearson r=0．524，P〈0．05，Spearman，rho=0．747，P〈0．01）。结论Akt及pAkt蛋白特异性过表达于胃癌组织，可能是蛋白翻译水平和（或）代谢水平改变的结果。Akt蛋白的磷酸化可能促进胃癌细胞的恶性转化和淋巴结转移，VEGFC可能在其中发挥重要的介导作用。     

15-羟基前列腺素脱氢酶与胃癌发生关系的研究

NAD依赖性15-羟基前列腺素脱氢酶（15-PGDH）是前列腺素和相关廿烷类生物降解、灭活的关键酶，其表达缺失或减低可能与一些恶性肿瘤的发生、发展密切相关。目的：探讨15-PGDH在胃癌组织和癌旁组织中的表达及其与临床病理特征的关系，初步评价15-PGDH在胃癌发生、发展中的作用及其意义。方法：随机收集30例胃癌患者的癌组织、癌旁3cm和6cm的对照组织，以及10例胃息肉、萎缩性胃炎和健康志愿者正常胃黏膜组织标本，以免疫组化和Western blot法检测15-PGDH蛋白表达，半定量逆转录聚合酶链反应（RT-PCR）法检测其mRNA表达，并分析其临床病理特征。结果：癌组织中15-PGDH蛋白和mRNA表达显著低于癌旁3cm、癌旁6cm对照组织和正常胃黏膜（P均〈0.01），约1／3癌组织中15-PGDH表达缺失，胃息肉和萎缩性胃炎组织显著低于正常胃黏膜（P均〈0．01）；癌组织中15-PGDH蛋白表达量较癌旁3cm和6cm对照组织分别平均降低5．7倍和8．3倍，胃息肉和萎缩性胃炎组织较正常胃组织分别平均降低2．0倍和2．1倍；黏液腺癌中15-PGDH蛋白量显著低于高分化腺癌（只〈0．05），印戒细胞癌中15-PGDH蛋白和mRNA的表达均缺失：伴有远处转移组15-PGDH蛋白和mRNA表达显著低于无远处转移组（P〈0．05和P〈0．01）；Ⅳ期胃癌组织15-PGDH蛋白表达量显著低于Ⅰ期（P〈0．01），其mRNA表达在Ⅲ、Ⅳ期胃癌组织中显著低于Ⅰ、Ⅱ期胃癌（P均〈0．01）。结论：15-PGDH在胃癌组织中表达减少甚至缺失，在癌旁组织和胃癌前状态中表达减少；15-PGDH表达缺失或减少可能是胃癌发生、发展．以及胃癌浸润转移的重要机制之一。     

BRMS1基因在胃癌中的表达及意义

目的观察乳腺癌转移抑制基因（BRMS1）在胃癌组织中的表达,并探讨其在胃癌侵袭转移中的作用及与胃癌生物学的关系。方法采用RT-PCR方法检测44例胃癌及相应癌旁正常胃组织中BRMS1的表达。结果 44例胃癌旁正常胃组织中有35例（79.5%）表达BRMS1,而胃癌组织中有14例（31.8%）表达,胃癌组织中BRMS1的表达水平明显低于癌旁正常胃组织（P〈0.01）,BRMS1的表达水平与胃癌的分化程度、浸润程度及淋巴结转移密切相关（P均〈0.05）。结论 BRMS1 mRNA在胃癌组织中表达降低,其可作为反映胃癌浸润、转移潜能的参考指标之一。     

胰腺癌组织DPC4／Smad4 mRNA表达的临床意义

目的：观察DPC4／Smad4 mRNA在胰腺癌组织中的表达及其临床意义。方法：应用原位杂交技术对23例胰腺癌组织中的DPC4／Smad4 mRNA表达进行研究。结果：胰腺癌组织中DPC4／Smad4 mRNA表达率为52．2％（12／23），低于癌周组织表达率9／10。统计学分析显示，DPC4／Smad4 mRNA表达与胰腺癌病理分级呈显著相关（P＜0．05），与胰腺癌临床分期有无局部淋巴结转移间无显著相关（P＞0．05）。结论：DPC4／Smad4 mRNA基因表达有可能作为胰腺癌诊断与鉴别诊断的生物学指标之一。     

蛋白翻译起始因子C_2在原发性肝细胞肝癌中表达的意义

目的研究C_2mRNA及其蛋白在原发性肝细胞肝癌(HCC)中的表达及意义。方法用原位杂交检测C_2mRNA住21例HCC及其癌旁组织中的表达,ABC免疫组化法检测C_2蛋白在60例HCC及其42例癌旁组织中的表达,Western blot法检测C_2蛋白在HCC及其癌旁组织中的表达。结果 21例HCC及其癌旁组织中,C_2mRNA阳性率分别占23.8％(5/21)和85.7％(18/21),60例HCC及42例癌旁组织中,C_2蛋白阳性分别占27.3％(17/60)和83.3％(35/42),27例肝硬变中,C_2蛋白阳性率为77.8％(21/27)。χ~2检验:C_2mRNA及其蛋白在HCC癌旁组织中表达明显高于癌组织(P<0.001);C_2蛋白在肝硬变中的表达明显高于HCC癌组织(P<0.01);C_2的表达与患者年龄、HBsAg及AFP有明显关系(P<0.05);而与癌组织的分化程度、肿瘤大小、淋巴转移无关;Western blot与免疫组化结果一致。结论 C_2基因表达下调可能与HCC的发生、发展及早期诊断有关。     

蛋白翻译起始因子C2在肝癌、胃癌中的表达与分布

目的 研究C2基因在肝细胞癌（HCC）、胃癌中的表达、分布及意义。方法 利用免疫组化法检测C2蛋白在60例HCC、58例胃癌组织中的表达，Western blot法检测C2蛋白在10例HCC及其癌旁组织中的表达。结果 60例HCC及42例癌旁组织中，C2蛋白阳性率分别为27．3％（17／60例）和83．3％（35／42例），C2蛋白在HCC癌组织中表达明显低于其癌旁组织（P＜0．001）；C2的表达与病人年龄、HBsAg及AFP明显关系（P＜0．05）。Western blot结果与免疫组化一致。在HBXAg阳性的49例HCC中，C2蛋白阳性率为20．4％，在HBXAg阴性的11例HCC中，C2蛋白阳性率为63．6％。C2在HBXAg阳性的HCC中的表达明显低于HBXAg阴性者（P＜0．05）。58例原发性胃癌及44例癌旁组织中，C2蛋白阳性率分别占41．4％（24／58例）和77．3％（34／44例）。C2蛋白在胃癌组织中表达明显低于其癌旁组织（P＜0．05），且与胃癌组织的分化程度、浆膜浸润、肿瘤大小有明显的关系（P＜0．05）。结论 C2基因表达下调可能与HCC、胃癌的发生、发展有关。HBXAg对C2的抑制作用可能与HCC的发生，发展有关。C2可能是一潜在的抑癌基因。     

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.     

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.     

Difference of gene expression profiles between esophageal carcinoma and its pericancerous epithelium by gene chip

AIM: To study the difference of gene expression between esophageal carcinoma and its pericancerous epithelium and to screen novel associated genes in the early stage of esophageal carcinogenesis by cDNA microarray. METHODS: Total RNA was extracted with the original single step way from esophageal carcinoma, its pericancerous epithelial tissue and normal esophageal epithelium far from the tumor. The cDNA retro-transcribed from equal quantity of mRNA was labeled with Cy5 and Cy3 fluorescence functioning as probes. The mixed probes were hybridized with two pieces of BioDoor 4 096 double dot human whole gene chip. Fluorescence signals were scanned by ScanArray 3 000 laser scanner and farther analyzed by ImaGene 3.0 software with the digital computer. RESULTS: (1) A total of 135 genes were screened out, in which 85 and 50 genes whose the gene expression levels (fluorescence intensity) in esophageal carcinoma were more than 2 times and less than 0.5 times respectively compared with the normal esophageal epithelium. (2) There were also total 31 genes, among then 27 and 4 whose expressions in pericancerous tissue were 2-fold up-regulated and 0.5-fold down-regulated respectively compared with normal esophageal epithelium. (3) There were 13 genes appeared simultaneously in both pericancerous epithelium and esophageal carcinoma, while another 18 genes existed in pericancerous epithelium only. CONCLUSION: With the parallel comparison among these three gene profiles, it was shown that (1). A total of 135 genes, Whose expression difference manifested as fluorescence intensity were more than 2 times between esophageal carcinoma and normal esophageal epithelium, were probably related to the occurrence and development of the esophageal carcinoma. (2). The 31 genes showing expression difference more than 2 times between pericancerous and normal esophageal epithelium might be relate to the promotion of esophageal pericancerosis and its progress. The present study illustrated that by using the gene chip to detect the difference of gene expression profiles might be of benefit to the gene diagnosis, treatment and prevention of esophageal carcinoma.     

Expression and distribution of protein translation initiation factor C2 gene in hepatocellular carcinoma and gastric cancer tissue

OBJECTIVE: To study the expression, distribution and significance of the C2 gene in hepatocellular carcinoma (HCC) and gastric cancer tissue. METHODS: The avidin‐biotin‐peroxidase complex (ABC) immunohistochemical method was used to detect the expression of C2 protein in 60 specimens of HCC and 58 specimens of gastric cancer tissue, and the western blot technique was used to detect the expression of C2 protein in 10 specimens of HCC and associated pericancerous tissue. RESULTS: In 60 specimens of HCC and 42 specimens of associated pericancerous tissue, the rates of C2 protein detection were 27.3 and 83.3%, respectively; the latter being significantly greater than the former (P < 0.001). The expression of C2 protein was significantly correlated with the patient's age, HBsAg and α?fetoprotein (AFP; P < 0.05). The findings from western blotting were consistent with those from immunohistochemical methods. The rate of C2 protein detection was 20.4% in 49 specimens of HCC that were hepatitis‐B virus encoded X antigen (HBxAg)‐positive, but 63.6% in 11 HCC specimens that were HbxAg‐negative. The rate of C2 protein detection was significantly lower in HbxAg‐positive tissue than in HbxAg‐negative tissue (P < 0.05). In 58 specimens of gastric cancer tissue and 44 specimens of associated pericancerous tissue, the rates of C2 protein detection were 41.4 and 77.3%, respectively. The rate was significantly higher in gastric cancer tissue than in associated pericancerous tissue (P < 0.05), and C2 protein detection was significantly correlated with pathological grading, tumor size and the depth of invasion (P < 0.05). CONCLUSIONS: Low expression of the C2 gene might be involved in the development of HCC and gastric cancer. The suppression of C2 by HBxAg may play an important role in the development of HCC. C2 may be a potential tumor suppression gene.     

非小细胞肺癌组织中EMIA—ALK融合基因与TYMSmRNA表达的关系

目的探讨非小细胞肺癌（nonsmall-celllungcancer,NSCLC）组织中棘皮动物微管样蛋白4-间变淋巴瘤激酶（echinodcriBmicrotubuleassociatedproteinlike4-anaplasticLymphomakinase．EMIA．ALK）融合基因与胸苷酸合成酶（Thymidylatesynthase,TYMS）mRNA表达的关系。方法应用实时荧光定量PCR方法检测257例NSCLC组织中EML4-ALK融合基因、TYMSmRNA的表达。结果非小细胞肺癌组织中EMIA-ALK融合基因阳性率占4．28％（11／257）,在不吸烟患者中较高（P〈0．05）;TYMSmRNA高表达占63．42％（163／257）。与未检测到EMIA-ALK融合基因阳性的非小细胞肺癌患者比较,EMIA-ALK融合基因阳性与TYMSmRNA表达水平相关（P〈0．05）。结论非小细胞肺癌组织中EMIA-ALK融合基因阳性患者TYMS倾向低表达,可能从-线化疗药的培美曲塞中受益。