GRP78在胃癌生长中的作用*

 目的： 探讨内质网分子伴侣葡萄糖调节蛋白78（GRP78）参与胃癌细胞生长的作用。方法: 回顾性分析34例胃癌及34例癌旁组织,利用组织微阵列技术,构建组织阵列,采用免疫组织化学技术SP法检测该阵列中GRP78蛋白在胃癌及其癌旁组织中表达情况。利用Western blotting检测人体胃癌组织、癌旁组织（肿瘤旁1 cm处）和正常组织（远离肿瘤≥10 cm处）中GRP78蛋白的表达情况。利用Western blotting检测人胃癌细胞SGC7901和过表达GRP78的SGC7901-H78细胞（稳定转染GRP78）中GRP78和生长相关蛋白cyclin D1的表达情况。结果: (1) 免疫组织化学检测发现：人体胃癌组织中GRP78表达水平明显高于癌旁组织和正常组织,且与性别、分化程度相关（P<0.05）; (2) Western blotting检测发现：胃癌组织中GRP78蛋白的表达明显高于癌旁组织和正常组织;(3)  Western blotting检测发现： 相对于SGC7901细胞,SGC7901-H78细胞中GRP78蛋白的表达明显升高,同时检测生长相关蛋白cyclin D1的表达发现,随着GRP78表达的上调,cyclin D1蛋白表达增加。结论: GRP78蛋白的高表达可能参与了胃癌细胞的生长。     

胃癌组织CDKN2A基因座p14ARF基因变异及意义

目的:探讨胃癌组织p14^ARF基因变异及其意义。方法: 应用PCR、PCR-SSCP、PCR甲基化分析法和RT-PCR分别检测48例胃癌及癌旁组织中p14^ARF基因纯合性缺失、突变、CpG岛甲基化及其mRNA表达状况。结果:①胃癌组织p14^ARF基因纯合性缺失率为31.3%(15/48), 癌旁组织均未见纯合性缺失。②33例无纯合性缺失的胃癌及癌旁组织均未见p14^ARF基因点突变。③胃癌组织p14^ARF基因甲基化率为47.9%(23/48), 癌旁组织仅2例甲基化, 两者差异有显著(P＜0.01)。④45.8%(22/48)的胃癌组织p14^ARFmRNA无表达。3例外显子E1β和E2共甲基化者p14^ARFmRNA均无表达(100%), 20例单纯E2甲基化者仅3例无表达(15%), 两者差异有显著(P＜0.05)。ARF基因失活多由纯合性缺失和5’CpG岛甲基化所致, 其表达缺失与胃癌的发生密切相关。     

DNMT在胃癌中的表达及DNMT抑制剂5-氮杂-2’-脱氧胞苷对胃癌的影响

目的: 研究DNA甲基化转移酶(DNMT)各亚型DNMT1、DNMT2、DNMT3A、DNMT3B和DNMT3L在胃癌与胃黏膜中的表达特点,并分析DNMT抑制剂5-氮杂-2’-脱氧胞苷对胃癌和胃黏膜细胞的影响。方法: 免疫组化法对60例胃癌与癌旁组织行5种DNMT表达研究。免疫荧光法和Western免疫印迹法对SGC-7901、MKN-45(胃癌细胞)和GES-1(胃黏膜细胞)行5种DNMT表达研究。噻唑蓝比色法和流式细胞术分析5-氮杂-2’-脱氧胞苷对SGC-7901、MKN-45和GES-1增殖、细胞周期和凋亡的影响。结果: 胃癌与癌旁组织均有5种DNMT不同程度表达,但胃癌组织DNMT1和DNMT3A表达显著低于癌旁组织(P<0.01),DNMT2和DNMT3L表达也低于癌旁组织(P<0.05),只有DNMT3B在胃癌与癌旁组织中无显著差异(P>0.05)。胃癌细胞(SGC-7901和MKN-45)与胃黏膜细胞(GES-1)亦有部分DNMT表达,除DNMT3B在胃黏膜细胞表达较强外,余DNMT在胃癌与胃黏膜细胞中无显著差异。5-氮杂-2’-脱氧胞苷对胃癌与胃黏膜细胞的增殖率、周期分布和凋亡率并无显著影响。结论: DNMT在胃癌中表达呈降低趋势,抑制DNMT对胃癌细胞的增殖和凋亡无显著影响。     

蛋白激酶WEE1在胃癌组织中的表达及其临床意义

目的　探讨蛋白激酶WEE1在胃癌组织中的表达及其与胃癌临床指标的相关性,并阐述其在胃癌发生发展中的作用。 方法　使用Oncomine数据库分析WEE1在胃腺癌和胃黏膜组织中的表达差异。选取100例胃腺癌和80例癌旁组织标本,通过免疫组织化学法和组织芯片技术检测WEE1蛋白在胃癌和癌旁组织中的表达水平。对免疫组织化学结果中染色的比例以及染色程度进行半定量分析,对半定量分析结果与胃癌临床指标进行相关性分析。使用RT-PCR和Western blotting检测WEE1在胃癌和癌旁组织中的mRNA和蛋白表达水平。 结果　WEE1在100例胃癌患者中的阳性率为86%,高表达86例,低表达14例。通过Oncomine数据库分析,蛋白激酶WEE1在胃腺癌中的表达明显高于胃黏膜组织中的表达（P<0.01）。免疫组化、RT-PCR和Western blotting均显示,WEE1在胃癌组织中的表达明显高于癌旁组织（P<0.01）;免疫组化还显示WEE1的表达量随着胃癌组织分期的增加而增加（P<0.01）。 结论　蛋白激酶WEE1在胃癌组织中高表达,提示其可作为胃癌治疗靶点之一。     

StathminsiRNA抑制人胃癌SGC-7901细胞裸鼠移植瘤的生长并诱导细胞凋亡

目的: 探讨微管解聚蛋白stathmin对人胃癌SGC-7901细胞裸鼠移植瘤的生长和细胞凋亡的影响及其可能的分子机制。方法: 将0.1 mL SGC-7901细胞悬液(5×10¹⁰/L)接种于裸鼠背部一侧皮下,建立人胃癌SGC-7901细胞裸鼠移植瘤模型,将其分为3组:PBS组(PBS治疗)、对照siRNA组(接种对照siRNA,终浓度为100 nmol/L)和stathmin siRNA组(接种stathmin siRNA,终浓度为100 nmol/L),治疗方式均为腹腔注射。连续2周观察荷瘤裸鼠的生长情况,免疫组织化学方法检测增殖抗原Ki-67的表达,采用TUNEL研究肿瘤组织中细胞的凋亡,Western blotting检测裸鼠肿瘤组织中stathmin、Bcl-2和Bax蛋白的表达。结果: Stathmin siRNA能明显抑制胃癌裸鼠移植瘤的生长(P<0.05),stathmin siRNA组中裸鼠移植瘤的重量明显低于PBS组和对照siRNA组(P<0.05)。免疫组化结果表明,stathmin siRNA组中Ki-67的增殖指数明显低于PBS组和对照siRNA组(P<0.05)。TUNEL结果显示,stathmin siRNA组的凋亡细胞数显著高于PBS组和对照siRNA组(P<0.05)。Western blotting结果表明,与PBS组和对照siRNA组相比,stathmin siRNA组中stathmin和Bcl-2蛋白的表达显著下调,而Bax蛋白表达明显上升(均P<0.05)。结论: Stathmin在胃癌细胞增殖和细胞凋亡的调控中发挥重要作用,本研究有望为胃癌分子靶向治疗提供理论依据。     

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。     

转录因子XBP-1在胃癌组织中表达及其临床相关性分析

目的:分析转录因子X盒结合蛋白1(X-box binding protein 1,XBP-1)在胃癌及癌旁正常组织的表达情况,并探讨其差异表达的临床意义。方法:分别采用免疫组化、real-time PCR及Western blot实验对胃癌及癌旁组织中XBP-1的表达进行检测;最后根据随访资料进行临床相关性分析及生存分析,以探讨XBP-1与胃癌发生发展的联系。结果:XBP-1蛋白和mRNA水平的表达在胃癌组织中较癌旁组织中表达明显降低(P0.05)。临床数据分析显示,XBP-1蛋白表达水平与胃癌分化程度有关(P0.05),分化程度越低,XBP-1蛋白表达水平越低。Kaplan-Meier生存分析发现,XBP-1低表达组患者预后不良。结论:XBP-1可能作为关键基因影响胃癌的发生发展及其预后。     

膜联蛋白A7在胃癌组织中的表达和意义

目的 检测膜联蛋白A7（ANXA7）在胃癌组织中的表达改变,探讨胃癌的发生机制。 方法 收集人胃癌组织55例及正常胃组织25例,细胞裂解液裂解组织细胞,蛋白定量试剂盒进行蛋白定量后,Western blotting法和Real-time PCR法分别于蛋白水平和基因转录水平检测ANXA7在胃癌组织和正常胃组织中的表达改变,并用免疫组织化学法对ANXA7在细胞中的定位和表达进行检测。 结果 ANXA7在胃癌中蛋白水平和转录水平的表达均显著增高,与正常组相比差异有显著性（P<0.05）; ANXA7在胃癌细胞中定位于细胞质。 结论 膜联蛋白A7在胃癌中的表达增高有可能是胃癌发生的机制之一。     

胃癌组织中TRAP1的表达及意义

目的：探讨TRAP1蛋白在胃癌组织中的表达及其与临床病理特征的关系。方法运用免疫组化PV法检测60例胃癌和20例癌旁组织中TRAP1的表达,采用Western blot法检测10例新鲜胃癌及癌旁组织中TRAP1的表达。结果免疫组化PV 法及Western blot 法检测结果均显示TRAP1在癌旁组织中极少表达,在胃癌组织中表达增高,差异有显著性(P <0．01),TRAP1表达与患者性别、年龄、分化程度、浸润深度等无关,与肿瘤大小、淋巴结转移、TNM 分期密切相关(P <0．01)。结论 TRAP1在胃癌中高表达,可能参与胃癌的发生、发展。     

人膝骨关节炎血清生物学标志物的筛选

目的: 比较膝骨关节炎患者与正常人血清蛋白质组的差异,筛选其能用于骨关节炎诊断、治疗或发病机制研究的血清生物学标志物。方法: 利用双向荧光差异凝胶电泳技术比较膝骨关节炎患者血清(4例)与健康志愿者血清(4例)蛋白图谱的差异,使用基质辅助激光解吸电离-飞行时间质谱及生物信息学相关技术对获得的差异蛋白进行鉴定和初步分析,并用Western blotting进一步验证所鉴定的蛋白。结果: 成功建立了骨关节炎血清差异蛋白质组学的研究方法;通过质谱分析初步鉴定出8个差异蛋白,其中5个蛋白在膝骨关节炎组表达上调,3个蛋白在膝骨关节炎组表达下调。Western blotting进一步验证得到α₂-巨球蛋白在膝骨关节炎组表达上调,与双向荧光胶内差异凝胶电泳技术联合质谱分析的结果一致。结论: 膝骨关节炎患者血清与正常人血清蛋白表达存在明显差异,其中α₂-巨球蛋白可作为骨关节炎潜在的疾病相关生物学标志物进一步研究,为骨关节炎的诊断、治疗和发病机制的研究提供新的线索和实验基础。     

Alterations of MUC1 and MUC3 expression in gastric carcinoma: relevance to patient clinicopathological features

AIMS: Several studies have reported conflicting and inconclusive results concerning the clinical relevance of mucin expression in gastric carcinoma. This study investigated the correlations between aberrant expression of mucins in gastric carcinoma and patient clinicopathological features. METHODS: The expression of MUC1, MUC2, MUC3, MUC5AC, and MUC6 was investigated immunohistochemically in gastric carcinoma (n = 46) in relation to patient clinicopathological features. RESULTS: All normal gastric mucosa samples expressed MUC1, MUC5AC, and MUC6. MUC1, MUC2, MUC3, MUC5AC, and MUC6 were expressed in 29, 31, 30, 18, and 21 of the 46 cases of gastric carcinoma, respectively. The number of cases expressing MUC1 was significantly higher (p < 0.01) in patients with a small tumour size (>/= 5 cm) and in patients in clinical stages I-II, compared with clinical stages III-IV (p < 0.05). Expression was significantly lower (p < 0.05) in patients exhibiting metastasis. The number of cases expressing MUC3 was significantly higher in patients in clinical stages III-IV (p < 0.05), and in those with serosal invasion (p < 0.05) or metastasis (p<0.01). No significant relations were found between MUC2, MUC5AC, MUC6, and clinical stage, metastasis, or tumour size. CONCLUSIONS: Membrane bound mucins MUC1 and MUC3 appear to be associated with the development of gastric carcinoma. Patients who maintained high immunoreactivity for anti-MUC1 antibody had a better prognosis, whereas those with an increase in anti-MUC3 immunoreactivity had a poorer prognosis, as judged by tumour size, serosal invasion, and metastasis. However, no correlation was found between MUC2, MUC5AC, or MUC6 and clinical prognosis.     

Paradoxical expression of maspin in gastric carcinomas: correlation with carcinogenesis and progression

Maspin, a serine protease inhibitor related to the serpin family, can suppress invasion and metastasis of malignancies. To clarify the role of maspin in the genesis and progression of gastric carcinomas, its expression pattern and level were studied by immunohistochemistry on tissue microarrays containing gastric carcinoma (n = 237), normal gastric mucosa (n = 23), intestinal metaplasia (n = 38), and adenoma (n = 42); and the findings were compared with clinicopathological parameters. Furthermore, maspin expression in the gastric carcinoma cell lines (HCG-27, MKN28, and MKN45) was examined by immunohistochemistry and Western blotting. We found that cytoplasmic and nuclear maspin expression paralleled each other (P < .05) and decreased from intestinal metaplasia, adenoma, and carcinoma to normal gastric mucosa (P < .05). A significant positive association was noted with depth of invasion, lymphatic invasion, lymph node metastasis, and TNM stage (P < .05) but not with sex or Lauren's classification (P > .05). Univariate and multivariate analyses indicated that expression of maspin correlated negatively with cumulative patient survival in gastric carcinoma (P < .05) but was not an independent factor in the prognosis. The 2 independent factors, depth of invasion and lymphatic invasion, influenced the relation between nuclear maspin expression and survival, whereas only depth of invasion correlated with cytoplasmic maspin. Our study indicated that maspin expression experiences upregulation in gastric precancerous lesions and then slight downregulation with malignant transformation. High expression may paradoxically promote invasion and metastasis of gastric carcinomas and could be considered a good marker for the pathobiological behaviors of gastric carcinomas.     

Expression of PRL-3 phosphatase in human gastric carcinomas: close correlation with invasion and metastasis.

OBJECTIVE: High expression of PRL-3, a protein tyrosine phosphatase, has been reported to be associated with metastasis of colorectal carcinoma. The aim of this study is to investigate the significance of PRL-3 expression in tumor progression and metastasis of gastric carcinoma. METHODS: The levels of PRL-3 mRNA expression in 8 gastric cancer cell lines were examined by RT-PCR. Ninety-four human gastric carcinomas and 54 matched lymph node metastases were employed in this study. The expression of PRL-3 was detected by immunohistochemistry and the relationship with clinicopathological parameters was analyzed. RESULTS: The expression of PRL-3 mRNA was clearly detected in 7 of 8 gastric cancer cell lines (87.5%) by RT-PCR. In tumor samples, PRL-3 expression was detected in 68% of primary gastric carcinoma (with nodal metastasis 81.5%, without nodal metastasis 50%; p = 0.004). The incidence of PRL-3 expression in lymph node metastasis was significantly higher than that in primary gastric cancers (p < 0.001). Moreover, PRL-3 expression was closely associated with lymphatic invasion (p = 0.002), extent of lymph node metastasis (p = 0.002) and tumor stage (p = 0.045). CONCLUSIONS: These results strongly suggest that PRL-3 expression may play a significant role in invasion and metastasis of gastric carcinoma. PRL-3 might be a novel molecular marker for aggressive gastric cancer.     

胃癌组织中CapG蛋白的表达及其与临床病理特征及预后的关系

目的检测巨噬细胞加帽蛋白(gelsolin-like actin-capping protein,CapG)在胃癌及癌旁组织(距离癌组织边缘5 cm)中的表达,探讨CapG蛋白表达与胃癌临床病理参数及预后的关系。方法收集125例胃癌石蜡包埋组织样本及癌旁组织30例。患者均为未行放、化疗的初诊患者。每例组织样本均行CapG免疫组化染色,并设置对照组。通过病理医师评分来确定CapG蛋白的表达量,探讨CapG蛋白表达与胃癌临床病理参数及预后的关系。结果 CapG蛋白在细胞质和细胞核中均表达,呈棕黄色颗粒状。癌旁组织中基本无表达,胃癌组织中的表达显著高于癌旁组织;胃癌组织中CapG蛋白表达与患者性别、年龄、肿瘤大小、肿瘤位置无显著相关(P均0.05),与浸润深度(P=0.044)、淋巴结转移(P=0.026)、远处转移(P=0.001)及AJCC分期(P=0.012)显著相关。Kaplan-Meier生存曲线显示,CapG蛋白高表达组患者的总体预后较低表达组差,且差异具有显著性(P0.001)。Cox回归分析显示,肿瘤位置、淋巴结转移、远处转移、AJCC分期、CapG蛋白表达情况是胃癌患者预后的独立危险因素。结论 CapG蛋白在胃癌组织中高表达,可能与胃癌的发生、发展有关,可作为胃癌预后判断的指标之一,有望成为胃癌治疗的潜在新靶点。     

共刺激分子B7-H1在胃癌中表达上调

目的了解胃癌患者胃黏膜组织中共刺激分子B7-H1的表达情况及其与肿瘤转移和预后的关系。方法应用流式细胞技术、免疫化学染色、免疫荧光染色与Western blot检测胃癌细胞系SGC-7901与新鲜切除的胃癌组织、癌旁组织及其远端正常胃黏膜组织中B7-H1的表达,并对相关数据进行相应的统计学分析,确定B7-H1表达水平与患者临床病理指标的关系。结果SGC-7901细胞中有B7-H1蛋白表达,主要分布在细胞膜,细胞质中少量;胃癌组织中B7-H1阳性表达率为(13/21)62%,癌旁组织中为(7/21)33%,而正常胃黏膜组织中没有B7-H1表达。统计分析显示,胃癌组织中B7-H1表达与肿瘤的浸润深度、周围淋巴结转移、远处转移及pTNM分期有关(P<0.05)。结论B7-H1分子可作为胃癌早期诊断与预后判断的新指标。     

食管癌中COX-2表达与其临床病理特征及预后的关系

目的：检测COX-2在食管癌中的表达，探讨其与食管癌临床病理特征及术后患者预后的关系。方法：应用SABC免疫组化染色法检测89例手术切除的食管癌组织和20例正常食管粘膜组织COX-2蛋白的表达，分析COX-2在食管癌组织的表达与患者性别、年龄、肿瘤部位、病变长度、浸润深度、区域淋巴结转移、远处转移、鳞癌分化程度的关系。并对其中81例有随访资料者，分析COX-2表达与预后的关系。结果：食管癌组COX-2呈阳性表达率为94.38%（84/89），且着色较深，正常食管组COX-2呈阳性表达率为60%（12/20），着色浅。食管癌中，随着肿瘤浸润深度的增加、鳞癌细胞分化程度的减低，COX-2的表达逐步增强（P＜0.05）。COX-2的表达与其它临床病理特征，包括性别、年龄、肿瘤生长部位、肿瘤大小、有无区域淋巴结转移、有无远处转移无相关（P＞0.05）。COX-2低表达组与高表达组的生存时间有显著差异，前者生存时间较后者为长（P＜0.01）。结论：COX-2在食管癌中的表达高于正常食管粘膜。肿瘤浸润越深、鳞癌分化程度越低，COX-2的表达越强；COX-2的表达与食管癌患者的年龄、性别、肿瘤生长部位、肿瘤长度、有无区域淋巴结转移、有无远处转移均无关。COX-2的表达与食管癌术后患者的预后相关，COX-2呈低表达者的生存时间比COX-2高表达者长。     

中国胃癌患者HER2蛋白表达与临床病理学参数及预后的关系

目的 评估中国胃癌人群中HEB2蛋白表达的流行病学情况,分析其与患者临床病理学参数之间的关系,探讨HEB2蛋白表达与胃癌患者预后的关系.方法 使用免疫组织化学方法检测胃癌患者中HER2蛋白的表达,以修订后HereepTest评分标准进行评分.x2检验分析HEB2蛋白表达与胃癌患者临床病理学参数之间的关系.生存分析应用Kaplan-Meier法,生存率比较采用Logrank 检验.结果 860例患者中位年龄59岁,男女比例为2.06∶1,其中进展期胃癌患者为774例.使用修订后的HercpTest评分标准评判HER2蛋白阳性表达(3+)的患者为77例(9.0%,77/860),其中进展期胃癌中的HER2阳性表达患者是69例,占进展期胃癌的8.9%(69/774).HEB2蛋白在胃癌中的阳性表达与肿瘤分化、Lauren分型及WHO分型相关,与年龄、性别、肿瘤发生部位及临床分期不相关.生存分析提示HER2蛋白阳性表达的胃癌患者生存率与阴性表达者无明显差异.结论 虽然胃癌HER2蛋白的阳性表达与肿瘤分化及组织学类型相关,但其可能不是胃癌患者的独立预后因子;HER2蛋白在胃癌中的阳性表达可能不影响胃癌患者的预后.

Abstract：

Objective To evaluate the epidemiological status of HER2 protein expression in Chinese patients with gastric carcinoma,and to study its clinical and prognostic significance and the association with the clinicopathological features.Methods The clinical data were reviewed in 860 patients with gastric carcinoma admitted to Guangdong General Hospital from 2003 to 2010.The HER2 status Was evaluated using immunohistochemistry(IHC).The modified Heroep Test scoring criterion Was used to assess HER2 protein expression.The association between HER2 expression and clinicopathological features was analyzed by X2 test.Kaplan-Meier analysis,log-rank test and Cox regression model were used for the survival analysis.Results The median age of the patients Was 59 years, and the male-to-female ratio was 2.06∶1.Positive expression of HER2 protein(3+)was found in 77(9.0%)cases of gastric carcinoma,and in 69(8.9%)edvanced gastric cancers.There waS significantly positive association between HER2 overexpression and tumor differentiation, Lanren claasification and WHO classification.No significant association Was observed between HER2 protein expression and patients'age,gender,tumor location and clinical stage.There waS no statistically significant difference in survival rate between patients with positive HER2expression and negative ones. Conclusion Though there WaS significantly positive association between HER2 expression status and tumor differentiation,histological type,it may be of limited prognostic value in gastric cancer patients.