slp-76基因在肝癌及其他消化系统肿瘤中的差异表达分析

目的分析slp-76基因在肝癌、食管癌、贲门癌、胃癌、结肠癌及其癌旁组织中的表达情况。方法应用微阵列技术筛选出slp-76基因在肝癌组织中高表达,以逆转录-聚合酶链反应(RT-PCR)技术对上述消化系统肿瘤组织及其癌旁组织中的slp-76基因表达进行检测;应用Northern blot技术对肝癌组织及其癌旁组织中的slp-76基因表达进行检测。结果 RT-PCR证实slp-76在79.3%(24/29)的肝癌组织中高表达(P=0.005);在73.3%(22/30)的食管癌组织中高表达(P=0.005);在75%(21/28)的贲门癌组织中高表达(P=0.014);在75%(15/20)胃癌组织中高表达(P=0.005);在72.2%(13/18)结肠癌组织中高表达(P=0.008);Northern blot证实slp-76基因在肝癌组织中呈高水平表达。结论 slp-76基因在消化系统肿瘤中高表达,slp-76与上述消化系统肿瘤生长关系密切;在RNA水平,该基因在上述肿瘤间的表达无显著性差异(P=0.968)。     

食管癌和贲门癌及胃癌的基因表达

为从基因角度探讨食管癌、贲门癌和胃癌的发病机理有无差别，用免疫组化抗生蛋白链菌素－生物素标记法（LSAB）检测34例食管癌、90例贲门癌及68例胃癌患肿瘤标本中抑癌基因－p53蛋白和增殖细胞核抗原（PCNA）的表达，结果表明PCNA的表达在三种癌变中均较高，食管癌与贲门癌p53的阳性率相近（76．47％，76．67％）而胃癌较低（45．59％），差异有统计学意义（P＜0．05）。     

食管癌组织环氧化酶-2的表达与血管生成的关系

目的:探讨环氧化酶-2(COX-2)在食管癌组织的表达及其与肿瘤血管生成的关系．方法:免疫组化法检测食管鳞癌手术切除标本90例和癌旁正常黏膜34例中COX-2表达,采用抗CD34抗体标记微血管内皮细胞,计算微血管密度(MVD)．分析COX-2表达与MVD及其与食管癌主要临床病理特征的相关性．结果:食管癌组织COX-2阳性表达率为84．4％显著高于癌旁正常黏膜的20．6％(x2=45．47,P =0．00)．COX-2表达与肿瘤细胞分化程度、临床TNM分期和淋巴结转移密切相关,TNM分期中Ⅲ Ⅳ期的食管鳞癌组织中COX-2表达率为92．9％,显著高于Ⅰ Ⅱ期的70．6％(x2= 7．99,P=0．005)．高、中分化的食管鳞癌组织中COX-2表达率为92．9％,显著高于低分化的 70．6％(x2=7．99,P=0．005)．伴有淋巴结转移的食管鳞癌组织中COX-2表达率为94．3％,显著高于无淋巴结转移的70．3％(x2=9．61,P= 0．002)．食管癌组织MVD值为29．68±3．81, 显著高于癌旁正常黏膜的15．12±2．80(t= 20．28,P=0．00)．MVD与肿瘤的TNM分期和淋巴结转移密切相关,TNM分期中Ⅲ Ⅳ期的食管鳞癌组织中MVD值为31．46±3．52,显著高于Ⅰ Ⅱ期的26．74±2．06(t=-7．09,P=0．00)．伴有淋巴结转移的食管鳞癌组织中MVD为 31．72±3．43,显著高于无淋巴结转移的26．76 ±2．01(f=-7．90,P=0．00)．Spearman等级相关分析表明,MVD与COX-2表达呈显著正相关(r =0．607．P=0．00)．结论:COX-2异常表达及其诱导的血管生成在食管癌的侵袭和淋巴结转移中起重要作用．     

黏着斑激酶在结肠癌组织中的表达及意义

目的探讨黏着斑激酶（FAK）在结肠癌发生、发展中的作用。方法采用RT-PCR法检测30例新鲜结肠癌及与之相对应的癌旁组织的FAK mRNA表达；同时用Western印迹法检测20例新鲜结肠癌及相对应的癌旁组织FAK蛋白表达水平，调平每对组织FAK蛋白含量后再行FAK Tyr397磷酸化位点蛋白检测。结果30例结肠癌组织FAK mRNA阳性率为90．0％，其表达值为0．745±0．530，对应癌旁组织阳性率为43．3％，其表达值为0．241±0．131（P〈0．01）；20例结肠癌组织FAK蛋白阳性率为95．0％，表达值为0．482±0．150；对应癌旁组织阳性率为60．0％，表达值为0．269±0．015；P均〈0．01。20例结肠癌组织中FAK Tyr397磷酸化蛋白阳性率为90．0％（18／20），表达值为0．385±0．021；而对应癌旁组织阳性率为20％（4／20），表达值为0．110±0．005，P均〈0．01。结论FAK特别是FAK Tyr397磷酸化蛋白表达增加在结肠癌的发生、发展中可能起重要作用。     

FHIT基因异常与消化道恶性肿瘤

目的探讨消化道恶性肿瘤细胞中三联脆组(fragile histidine triad,FHIT)基因的异常转录情况.方法应用巢式逆转录聚合酶链反应(nested RT-PCR)对96例消化道恶性肿瘤(21例食管癌、43例胃癌和32例结肠癌)及相应的癌旁组织,18例正常组织FHIT基因cDNA片断进行扩增.结果在33.3%的食管癌组织,51.2%的胃癌组织和31.3%的结肠癌组织中检出异常FHIT转录,而在相应的食管、胃、结肠癌旁组织中FHIT异常转录检出率分别为4.8%、20.9%和9.4%,二者差异有显著性(P＜0.05);18例正常组织中未见FHIT异常转录.结论食管癌、胃癌和结肠癌组织FHIT基因的异常转录可能与消化道恶性肿瘤的发生、发展有关.     

SLP-2在胃癌中的表达及意义

目的:研究SLP-2基因在胃腺癌及正常胃黏膜中的表达情况.方法:应用免疫组织化学法及RT-PCR法分别检测45例及40例胃腺癌及其癌旁正常胃黏膜中SLP-2基因的表达,并结合免疫组织化学结果及胃癌患者的临床病理资料进行分析.结果:免疫组织化学和RT-PCR结果均显示SLP-2在胃腺癌组织中的表达高于正常配对胃黏膜(68.9%vs26.7%,1.12±0.47vs0.63±0.31,均P<0.01),且SLP-2的表达与胃癌TNM分期及有无淋巴结转移相关(2=5.32、4.78,均P<0.05).结论:SLP-2基因在胃腺癌组织中高表达,可能参与胃腺癌的发生发展和转移.     

Syndecan- 1、MMP-9在胃癌组织中表达的相关性及其临床意义

用免疫组织化学染色法检测Syndecan-1和基质金属蛋白酶-9（MMP-9）在55例胃癌组织及癌旁3cm、癌旁10cm组织中内的表达情况。结果癌旁胃癌3cm及10cm组织中Syndecan-1阳性表达率分别为14．55％、29．09％、94．55％，MMP-9组织的阳性表达率分别为92．73oA，81．82％、16．36％；Syndeean-1与MMP-9表达在胃癌及癌旁10cm组织中呈负相关r=-0．480，P=0．000；r=-0．543，P=0．000）。提示胃癌组织中Syndecan-1呈低表达，MMP-9呈高表达，两者呈负相关；二者可能对肿瘤进展及恶性程度的判断具有重要意义。     

生存素基因在胃癌组织中的表达及其与胃癌预后关系

目的探讨生存素(survivin)基因在胃癌组织中的表达及其与胃癌预后关系。方法病例来源于1995年7月至1996年6月中山医院住院手术患者，采用免疫组化Envision二步法检测手术切除的胃癌病灶组织中生存素基因表达。根据手术病理诊断与临床表现确定肿瘤TNM分期。所有病例随访至少5年或直到死亡。比较不同病理类型、不同TNM分期生存素表达差异。按生存素基因是否表达比较两组患者生存曲线的差异。结果共有96例患者进入研究和随访观察，其中男59例，女37例；年龄29～84岁，平均56岁，68例表达生存素基因产物，阳性率70．8％。病理类型中腺癌78例，非腺癌18例，不同病理类型生存素基因表达相似(腺癌69．7％、非腺癌60．0％，P=0．369)。对腺癌病例分析显示：低分化组生存素基因表达高于中高分化组(82．1％比62．5％，P=0．053)；肿瘤累及固有肌层组和全层组生存素基因表达高于肿瘤局限于黏膜或黏膜下层组(81．3％，75．9％比33．3％，P=0．020)。生存素表达与淋巴结转移与否无关(68．1％比70．8％，P=0．771)。生存素阳性组5年生存率(42．93％)低于生存素阴性组(52．93％)，但差异无统计学意义。结论生存素基因在胃癌中高表达。生存素基因表达与胃腺癌分化程度以及肿瘤累及深度有关，需进行更多研究评价其在预后中的作用。     

食管、贲门癌组织中CMTM4、8的表达及意义

目的检测食管、贲门癌组织中人类趋化素样因子超家族（CKLFSF）成员CMTM4、8的表达并探讨其临床意义。方法使用组织微阵列（TMA）和免疫组织化学方法（IHC）检测60例食管癌和贲门癌肿瘤组织以及癌旁正常组织中CMTM4、8蛋白的表达。结果CMTM4、8在癌旁正常组织中的阳性表达率均为100％,在肿瘤组织中的阳性表达率分别为31．7％和26．7％,两组相比,P〈0．01。CMTM4、8蛋白在肿瘤组织中的表达程度与患者性别、病理类型、病理分化程度和临床分期之间无明显相关（P〉0．05）。结论CMTM4、8可能是肿瘤抑癌基因。     

人结肠癌组织中肿瘤耐药相关基因的表达及其临床意义

背景：人结肠癌对化疗药物的耐药是其化疗失败的主要原因，一些与肿瘤耐药相关的基因从中起着一定作用，有关耐药相关基因在人结肠癌中表达水平的联合检测报道较少。目的：探讨人结肠癌组织中耐药相关基因p21（^WAF1/CIP1）、MRP2、BRCA2、CYP3A5、BCRP和ELK-1转录水平的表达及其临床意义。方法：对30例结肠癌手术标本的癌组织、癌旁组织和正常组织分别以逆转录聚合酶链反应（RT—PCR）法检测肿瘤耐药相关基因p21^（WAF1/CIP1）、MRP2、BRCA2、CYP3A5、BCRP和ELK-1转录水平的表达．比较它们存不同组织中表达的差异及其与肿瘤临床特点之间的关系。结果：结肠癌组织中，MRP2转录表达水平显著高于正常组织（P=0．037）；CYP3A5转录表达水平显著低于正常组织（P=0．042）；ELK-1转录表达水平显著高于癌旁组织和正常组织（P=0．016和P=0．022）。在结肠癌组织和正常组织中，肿瘤耐药相关基因转录水平表达两两之间相关性有不同。结肠癌组织中．肿瘤耐药相关基因与结肠癌临床特点之间均无相关性。结论：在结肠癌的发生和耐药过程中，肿瘤耐药相关基因MRP2、CYP3A5和ELK-1起了一定作用．而p21^（WAF1/CIP1）、BRCA2和BCRP基因的作用可能不明显。     

人结肠癌中凋亡抑制基因Survivin的表达

背景:细胞增殖和凋亡失衡将导致肿瘤的发生,并直接影响肿瘤的生物学行为。survivin是一种新的凋亡抑制基因,在大多数肿瘤中显著高表达。目的:探讨survivin基因在人结肠癌发生、发展中的作用,及其与p53基因的表达和结肠癌生物学行为的关系。方法:采用逆转录聚合酶链反应(RT鄄PCR)检测33例结肠癌组织及其相应癌旁组织和正常结肠黏膜中survivinmRNA和p53mRNA的表达,分析survivinmRNA的表达与p53mRNA的表达和结肠癌生物学行为的相关性。结果:69.7%的结肠癌组织中有survivinmRNA表达,表达率显著高于癌旁组织和正常结肠黏膜(42.4%和21.2%,P<0.01);survivin表达阴性癌组织的相应癌旁组织和正常结肠黏膜无一例有survivinmRNA表达。结肠癌组织中survivinmRNA的表达值显著高于癌旁组织和正常结肠黏膜(P<0.01)。33例结肠癌组织中仅2例p53mRNA表达缺失,癌旁组织和正常结肠黏膜中均有p53mRNA表达。结肠癌组织中p53mRNA的表达值与癌旁组织和正常结肠黏膜无显著差异;survivin表达阳性癌组织中p53mRNA的阳性表达率和表达值与survivin表达阴性组无显著差异。survivinmRNA和p53mRNA的表达与结肠癌的生物学行为无显著相关性。结论:survivin基因在人结肠癌组织中表达上调,提示其可能通过抑制结肠癌细胞凋亡,在结肠癌的发生、     

Different expressions of sialyl Tn antigen between polypoid and flat-type early colorectal cancers

PURPOSE: Sialyl Tn (STn) antigen is a cancer-associated carbohydrate antigen expressed in cancers of the digestive tract. We compared the proportion of specimens of flat-type colorectal cancers expressing STn with that of polypoid cancers, by examining the immunohistochemical reactivity of STn in various morphologic types of early and advanced colorectal cancers. METHODS; A total of 111 biopsies from the colorectal area were examined for STn expression, including 11 adenomas, 58 early cancers, and 42 advanced cancers. Each section was stained immunohistochemically for STn antigen. In each section, we examined STn expression in the cancer area, adjacent mucosa, and normal epithelium. RESULTS: STn expression was detected in 90.9 percent of adenomas, 36.2 percent of early cancers (T1), 64.3 percent of advanced cancers (>T1), and 52 percent of mucosa adjacent to cancer. The morphology of cancer tissue did not influence the number of specimens exhibiting STn antigen expression in mucosa adjacent to cancer cells. STn antigen was rarely expressed in flat or depressed-type early cancers (T1; 7.1 percent), and the expression was higher in moderately than in well-differentiated adenocarcinomas. In advanced cancers (>T1), a similar proportion of protruding and small ulcerative cancers expressed STn. CONCLUSION: Our results suggest that the low expression of STn antigen in flat-type cancers may be the result of different mechanisms of cellular transformation during carcinogenesis from the usual adenoma-carcinoma sequence in colorectal neoplasms.     

Upregulated expression of S100A6 in human gastric cancer

OBJECTIVE: The expression of S100A6 (calcyclin), a member of the S100 calcium binding protein family, is elevated in a number of malignant tumors, but there have been few reports about its expression in gastric cancer. The aim of this study was to investigate its expression regulations in human gastric cancer and noncancerous mucosa, and the response to chemotherapeutic drugs in the gastric cancer cell line. MATERIALS AND METHODS: In one matched gastric cancer sample pair, the serial analysis of gene expression (SAGE) experiment was conducted to compare the gene expression profiles between cancerous and adjacent tissues. To detect the expression regulations among more cancerous tissues, microarrays were carried out and real-time RT-PCR was conducted to validate the results. At the protein level, Western blot and tissue microarray (TMA) examination were further used to verify S100A6 expression. The regulation detection of S100A6 with flurouracil and doxorubicin at the mRNA and protein level was performed in the SGC7901 cell line. RESULTS: With the SAGE strategy, five times more S100A6 tags were identified in cancer tissues than in normal tissues. With the cDNA microarray, S100A6 was found to be significantly upregulated in 21 of 42 (50%) nonselective gastric cancers. In 10 other paired samples, the upregulation of S100A6 was consolidated with RT-PCR and Western blot analysis as well. A total of 14 endoscopy-sectioned gastric noncancerous lesions and corresponding normal gastric mucosa were also applied to profile the gene expression; both cDNA microarray and RT-PCR demonstrated no significant alterations of S100A6 at the mRNA level. TMA examination showed that 34 of 52 (65.4%) cancer samples were positively stained, while only 17 of 80 (21.3%) noncancerous lesions were positively detected and all nine normal mucosae were detected to be negative. An in vitro experiment showed that in the gastric cell line SGC-7901, S100A6 mRNA was detected to be upregulated from 24 to 72 h after treatment with 5 mg/L 5-flurouracil or 0.3 mg/L doxorubicin, and there were two wave upregulations of the S100A6 protein. CONCLUSION: The observed regulated expression of S100A6 suggests that it is associated with gastric cancer tumorigenesis and quantitation of S100A6 is a promising tool for diagnosis of gastric cancer.     

Induction of pS2 and hSP genes as markers of mucosal ulceration of the digestive tract

The recently discovered pS2 protein is expressed under estrogen control in a subset of estrogen receptor-positive breast cancers and in an estrogen-independent manner in normal stomach mucosa. The pS2 gene belongs to a family of genes encoding peptides that contain a conserved 5-cysteine domain, the P domains. Although the function of the pS2 protein is unknown, it has been suggested that it may have cell growth stimulatory activity. We report here that expression of the pS2 gene in the digestive tract, which is normally restricted to the stomach, is strongly induced by mucosal ulcerations elsewhere in the tract, most notably in Crohn's disease. pS2 gene expression is restricted to the mucosal layers adjacent to the ulcerations, in a region where a novel epidermal growth factor-secreting cell lineage was shown to be induced by mucosal ulceration. The human hSP gene, which contains a tandem duplication of the pS2 gene P domain and is coexpressed with the pS2 gene in normal stomach mucosa but not in breast cancers, is also expressed in Crohn's disease. We suggest that pS2 gene expression may provide a useful marker for mucosal ulcerations of the digestive tract.     

Analysis of differential gene expression patterns in colon cancer and cancer stroma using microdissected tissues

BACKGROUND AND AIMS: The surrounding cancer stroma is increasingly recognized as playing an important role in cancer proliferation, invasion, and metastasis. Here, we analyzed patterns of gene expression in colon cancer cells, surrounding stroma, and normal mucosa and normal stroma using laser capture microdissection. METHODS: Tissues were fixed from 11 patients undergoing colorectal resection for cancer, and laser capture microdissection was performed. Samples (4 per patient) were extracted for RNA, which was then used for focused gene arrays. In addition, protein expression of selected genes was determined by Western blot or immunohistochemistry. RESULTS: We showed differential expression patterns in cancer cells and surrounding stroma compared with their normal counterparts. Genes contributing to angiogenesis, cell cycle regulation, and proliferation were significantly increased in cancer cells compared with the adjacent normal mucosa. Genes contributing to angiogenesis, cancer invasion, and metastasis were significantly increased in surrounding cancer stromal cells compared with normal stroma. CONCLUSIONS: Delineating the differential patterns of gene expression between colon cancers and the surrounding reactive stroma will better determine the role of the stromal components in the progression of colon cancers and may lead to chemopreventive therapy targeted specifically to the stroma.     

大肠癌相关新基因SNC73在人上皮肿瘤组织中的表达研究

目的 原位检测大肠癌相关新基因SNC73在常见上皮肿瘤中的表达状况，探讨该基因表达与上皮肿瘤发生的相互关系。方法 采用cRNA/mRNA原位杂交的方法，用体外转录制备的地高辛标记的cRNA探针对大肠癌，胃癌，肝癌，肺癌和乳腺癌组织及同一患者的相应正常组织作表达情况的配对研究。结果 SNC73基因在胃，大肠黏膜的上皮细胞及淋巴细胞呈强阳性表达，在其他组织细胞中未见表达；在大肠癌和胃癌组织中存在明显的表达降低或缺乏；在肝癌，肺癌和乳腺癌及其相应正常组织中都未见表达。结论 SNC73是一个IgA样消化道肿瘤相关基因，可作为候选的抑癌基因进行深入研究。     

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.