HCA587 antigen expression in normal tissues and cancers: correlation with tumor differentiation in hepatocellular carcinoma

The HCA587 gene, identified by serological analysis of recombinant cDNA expression library (SEREX) from a hepatocellular carcinoma (HCC) patient, encodes a new member of cancer-testis antigens. HCA587 mRNA expression in normal tissues and cancers has been previously reported. To estimate its immunogenicity to induce immune response, it is essential to analyze HCA587 expression at the protein level. In this study anti-HCA587 polyclonal antibody, termed "TC-1," was generated, and the expression of HCA587 protein was assessed by immunohistochemical staining in a panel of normal and tumor tissue sections. No HCA587 protein was shown in normal tissues except germ cells in testis and Purkinji cells in cerebellum. In HCC specimens the HCA587 protein was expressed in 37.1% (26 of 70) samples. The expressed protein was either located in the cytoplasm or nucleus depending on the individual samples. More importantly, there appears to be correlation between the tumor differentiation of HCC and HCA587 protein expression, ie, the lower differentiation, the higher percentage of protein expression. Coincidentally, seroreactivity showed that the Ab specific to recombinant HCA587 protein was detected only in the sera of three patients with poorly differentiated HCCs. HCA587 antigen was also expressed in different proportions in melanoma, lymphoma, pancreatic cancer, and lung cancer.     

肝癌组织中MAGE-A1表达及其临床意义

目的检测黑色素瘤抗原-A1(MAGE—A1)在肝癌组织中表达,探讨MAGE—A1基因与肝细胞肝癌(HCC)患者临床病理指标的关系。方法应用RT—PCR法检测31例HCC组织及相应癌旁组织中MAGE—A1基因的表达。应用组织芯片和免疫组化技术分析178例HCC组织及相应癌旁组织中MAGE—A1抗原表达。结果31例HCC组织标本中有21例(67．74％)表达MAGE—A1基因,而相应的癌旁组织MAGE—A1基因均不表达。178例HCC组织中95例表达MAGE—A1抗原(53．37％),相应的癌旁组织MAGE—A1抗原均不表达。结论MAGE—A1基因在HCC组织中有较高表达,其阳性率与HCC患者的临床病理指标无相关关系。     

EZH2在肝细胞癌中的表达及临床意义

目的 探讨EZH2在肝细胞癌中的表达及临床意义方法采用RT-PCR、免疫组织化学和Western blotting方法分析肝癌及癌旁组织EZH2mRNA转录和蛋白表达水平及其与临床病理因素的关系.结果 在42例肝癌组织中检出26例EZH2mRNA,而在相应的42例癌旁组织中只检出12例,癌组织EZH2mRNA阳性率高于...     

Histological study of PIVKA-II expression in hepatocellular carcinoma and adenomatous hyperplasia

Although serum concentration of protein induced vitamin K absence or antagonist II (PIVKA-II) has been widely used for diagnosing hepatocellular carcinoma (HCC), little information is available concerning tissue PIVKA-II as an immunohistochemical marker for liver histology. In this study, we examined the expression of PIVKA-II in precancerous nodules (adenomatous hyperplasia) and various differentiation grades of HCC by immunohistochemical study using the monoclonal anti-PIVKA-II antibody (MU-3). We examined the relationship between tissue PIVKA-II staining and serum PIVKA-II level, tumor histology and tumor size. PIVKA-II was mainly detected in the cytoplasm of the HCC cells. The positive rates of PIVKA-II were as follows: adenomatous hyperplasia (AH), 0% (0/9); well-differentiated HCC, 65% (15/23); moderately differentiated HCC, 85% (22/26); poorly differentiated HCC, 54% (7/13). The expression of tissue PIVKA-II staining in moderately differentiated HCC was significantly higher than in well- or poorly differentiated HCC, whereas the serum PIVKA-II level in poorly differentiated HCC was higher than well- or moderately differentiated HCC. There was no relationship between the expression of PIVKA-II in cancer tissues and serum levels of PIVKA-II. Immunohistochemical studies revealed that PIVKA-II was expressed even in small-sized or well-differentiated HCC cells, but expression was not detected in AH. It was concluded that PIVKA-II is a useful immunohistochemical marker, even in small-sized or well-differentiated HCC.     

Expression of HNF-1α and HNF-1β in various histological differentiations of hepatocellular carcinoma

Hepatic nuclear factor 1 (HNF-1) regulates genes in a hepatocyte-specific manner. It has been previously reported that the ratio of HNF-1α and HNF-1β mRNA is related to histological differentiation hepatocellular carcinoma (HCC). In this study, the expression levels of the HNF-1α and HNF-1β proteins were analysed relatively and quantitatively in various histologically differentiated HCC and surrounding non-cancerous tissues, and HNF-1α binding activity for the AT element of the B domain of the human α-fetoprotein enhancer was examined. Western blot analysis demonstrated that HNF-1α protein was expressed at a higher level in well-differentiated HCC tissues than in the surrounding non-HCC tissues; on the other hand, the HNF-1α protein was expressed at lower levels in moderately and poorly differentiated HCCs than in the surrounding non-HCC tissues. The levels of HNF-1β expression in well-differentiated and poorly differentiated HCCs were similar to and higher than those found in the respective surrounding non-cancerous portions. In binding assays, HNF-1 binding activity was high in well-differentiated HCC and lower in moderately and poorly differentiated HCCs. Most well-differentiated HCC cases showed immunohistochemical expression of HNF-1α. These findings show that poor histological differentiation of HCC correlates with decreases in the level and activity of HNF-1α proteins. © 1998 John Wiley & Sons, Ltd.     

GPC3C末端融合蛋白在肝细胞癌中的分布及其临床意义

目的：检测Glypican-3(GPC3)在肝细胞癌中的表达。探讨GPC3在肝细胞癌中的作用。方法：（1）定向克隆GPC3基因C末端180bp的片段,IPTG诱导原核表达载体pGEX-5X-1、鉴定并利用谷胱苷肽转移酶纯化GPC3末端融合蛋白;（2）免疫家兔,制备抗体;（3）应用免疫组化（SP法）检测了肝细胞癌及其癌旁组织中GPC3的表达。结果：（1）纯化了GPC3末端融合蛋白;（2）制备了抗人GPC3多克隆抗体;（3）免疫组化结果表明25例低分化肝细胞癌中18例为阳性,表达率为72%;阳性结果为棕色颗粒均匀分布于肝细胞癌的细胞膜上和胞浆内;正常肝细胞,胆管细胞,血管内皮细胞、Ito细胞及成纤维细胞均呈阴性;癌旁组织不表达,GPC3的表达和肿瘤大小及门脉癌栓形成有密切关系。结论：GPC3在肝细胞癌中高表达,且具有细胞特异性;GPC3在低分化的肝细胞癌中有重要作用。     

Relationship between p53 overexpression and the proliferative activity in hepatocellular carcinoma

Relationship between p53 protein overexpression and clinicopathological findings and the proliferative activity was studied in 50 cases of hepatocellular carcinoma (34 biopsy and 16 surgically resected cases) using immunohistochemistry. Overexpression of p53 was observed in 26.5% of biopsy cases and 31. 3% of surgically resected cases. Investigation of the relationship between the p53-positive rate and the clinical stage of HCC showed that it was significantly higher in Stage IV (the most advanced cancer; 54.5%) than in Stage I/II/III (13.0%) (p<0.05). Examination of the relationship between the p53-positive rate and tumor differentiation in the biopsy cases showed that p53 was positive in 9.1% of well differentiated carcinomas, 21.4% of moderately differentiated carcinomas, and 55.6% of poorly differentiated carcinomas, indicating that p53 positivity increased as tumors became less differentiated. The p53-positive rate of poorly differentiated carcinoma (55.6%) was significantly higher than that of well and moderately differentiated carcinoma (16.0%) (p<0.05). In the surgically resected cases, p53 overexpression tended to be more frequent in the less differentiated parts of each tumor nodule. In cases with nodule in nodule pattern of HCC, the p53-positive rate was different among nodules with the same level of differentiation. Examination of tumor cell proliferative activity using the proliferating cell nuclear antigen L.I. showed that this indicator was significantly higher in the p53-positive tumors than in the p53-negative tumors (52.7+/-32.4% vs. 32.4+/-15.3%: p<0.05). These results suggest that p53 overexpression may be involved in determining the dedifferentiation and the proliferative activity of HCC. Examination of the surgically resected cases confirmed that p53 overexpression became heterogeneous during the multistep carcinogenesis and growth process of HCC, which is considered to develop from a single cell. This finding suggests that p53 overexpression may be involved in tumor progression.     

CUEDC2在肝细胞癌组织中的表达及其临床预后的意义

目的:检测含CUE结构域蛋白2(CUEDC2)在肝细胞癌组织中的表达,探讨其与临床预后的关系。方法:选取具有完整临床及预后资料的186例手术切除肝细胞癌组织的石蜡组织标本,采用免疫组化技术检测CUEDC2的表达,并结合肝癌临床病理因素及预后进行分析。结果:CUEDC2在85.5%(159/186)的肝癌组织中有表达,其中低表达为52.2%(97/186),高表达为47.8%(89/186);CUEDC2的表达状态与血清甲胎蛋白(AFP)水平、肿瘤大小、肿瘤数目、肝癌分化程度和TNM分期有关(P0.05)。Kaplan-Meier生存分析显示CUEDC2高表达的患者术后总生存及无复发生存期较低表达者显著缩短;多因素Cox回归分析显示CUEDC2的表达状态、血清AFP和肿瘤数目是影响肝癌患者术后无复发生存(RFS)及总生存期(OS)的独立危险因素。结论:CUEDC2在多数肝癌组织中有表达,与肿瘤的生长、分化及预后有关;CUEDC2可以作为一个有价值的肝癌预后预测的新指标。     

Survivin在肝细胞癌中的表达特点及其与预后的关系

目的: 研究原发性肝细胞癌组织中Survivin的表达及其与肝癌细胞的生物学行为及预后的关系。 方法: 应用免疫组织化学染色对83例肝癌及相应癌旁组织中Survivin蛋白的表达情况进行检测，应用半定量RT-PCR技术对11例肝癌及相应癌旁组织中Survivin mRNA的表达情况进行检测，结合临床病理资料分析。 结果: 肝癌及癌旁组织中Survivin蛋白的表达率分别为63.9%(53例)和39.6%(21例)。肝癌组织中，41.5%(22例)表达于肝癌细胞核，45.3%(24例)表达于肝癌细胞浆，13.2%(7例)在胞核、胞浆中均有表达。统计学分析表明，Survivin蛋白阳性反应定位于胞核的病例，其肿瘤的包膜侵犯率和转移率更高 (P<0.05)；且术后生存期<2年的肝癌组中的核表达率显著高于术后生存期≥2年者(P<0.01)。RT-PCR结果显示，11例肝癌组织均表达Survivin mRNA，而癌旁组织只有45.5%(5例)表达。 结论: Survivin在肝癌中呈现高表达，其核表达与肝癌的恶性生物学行为及预后密切相关。     

Autoantibody response against a novel testicular antigen protein highly expressed in testis (PHET) in SSc patients

Systemic sclerosis (SSc) is characterized by excessive fibrosis and autoantibody production. However, the pathogenesis of SSc is still under investigation. We have demonstrated that a novel testicular antigen, protein highly expressed in testis (PHET), is overexpressed in SSc dermal fibroblasts and targeted by autoantibodies. PHET was identified by screening of HepG2 cDNA library using an SSc serum and was found to belong to UniGene cluster of sperm associated antigen 9 (SPAG9) from its nucleotide sequence. PHET mRNA expression was examined by RT-PCR using mRNA panels of human tissues, and PHET mRNA was highly expressed only in testis in normal tissues. Anti-PHET antibodies were detected in 8.4% of sera of SSc patients by immunoblotting, and associated with diffuse scleroderma and lung involvement. Expression of PHET mRNA and protein was increased in cultured SSc dermal fibroblasts compared with control fibroblasts. These results suggest that ectopic expression of PHET in dermal fibroblasts induces autoantibody production against PHET in patients with SSc.     

SCNM1在乙肝相关性肝癌中的表达及临床意义

目的:研究钠离子通道调节蛋白1(SCNM1)在乙肝相关性肝癌中的表达情况,并探讨其表达差异与临床病理特征及预后的关系。方法:标本来源于2013年1月～2015年12月在中山大学附属第三医院接收治疗的108例乙肝相关性肝癌患者,所有患者均签署知情同意书,符合医学伦理学规定。结合公共数据库中的肝癌资料,分析SCNM1的m RNA表达水平;应用RT-qPCR检测乙肝相关性肝癌组织及癌旁组织中SCNM1的m RNA表达水平,并分析SCNM1的表达水平与乙肝相关性肝癌临床病理特征的关系;利用Kaplan-Meier plotter分析SCNM1与肝癌患者预后的相关性。结果:TCGA数据库、Human Protein Atlas数据库和Oncomine数据库的分析结果显示,SCNM1在肝癌组织中的表达量明显高于正常肝组织(P 0. 01)。SCNM1主要分布于细胞核中。RT-qPCR检测结果显示,乙肝相关性肝癌组织中SCNM1的m RNA中位表达水平明显高于其配对癌旁组织(t=8. 082,P 0. 01)。乙肝相关性肝癌患者中SCNM1的表达水平与肝硬化、丙氨酸转氨酶和肿瘤大小具有相关性(P 0. 05),而与患者的性别、年龄和肿瘤包膜等临床病理特征无相关,SCNM1高表达的肝癌患者总生存时间较低表的患者达短(HR=1. 53,P=0. 016),SCNM1高表达的乙肝相关性肝癌患者总生存时间更短(HR=2. 41,P=0. 015)。结论:在乙肝相关性肝癌中,SCNM1高表达并且和患者的预后相关。SCNM1在乙肝相关性肝癌的发生中可能起着重要作用。     

eEF1A2在原发性肝细胞癌的表达

 目的: 研究真核翻译延长因子1A2(eEF1A2)在原发性肝细胞癌(HCC)组织的表达,以及eEF1A2过表达对HCC细胞生物学行为的影响。方法: 应用real-time PCR法和免疫组化法分别检测62例HCC癌组织与配对癌旁组织、20例正常肝脏组织eEF1A2的mRNA和蛋白表达,应用real-time PCR法与Western blot法分别检测几种HCC细胞株eEF1A2的mRNA与蛋白表达。构建GV287-eEF1A2表达慢病毒感染低表达eEF1A2的HCC细胞,应用real-time PCR法和Western blot法分别检测细胞eEF1A2 的mRNA和蛋白表达;应用MTT法、DNA倍体法和real-time PCR法分别检测细胞活力、细胞周期和白蛋白mRNA的表达。结果: HCC癌组织eEF1A2的mRNA表达水平和蛋白表达阳性率明显高于配对癌旁组织与正常肝脏组织(P<0.01);eEF1A2的mRNA和蛋白在HCC细胞株SMMC-7721和BEL-7402高表达,在SK-HEP-1低表达。构建的GV287-eEF1A2表达慢病毒可感染SK-HEP-1细胞使eEF1A2过表达;与阴性对照组相比,GV287-eEF1A2组SK-HEP-1细胞的活力升高,白蛋白的mRNA表达水平降低,G₀/G₁ 期的细胞比例显著减少,而S期和G₂/M期的细胞比例显著升高。结论: eEF1A2在HCC癌组织存在异位高表达;eEF1A2可能是HCC的一种潜在癌蛋白,其过表达可增强HCC细胞的增殖能力,降低HCC细胞的分化程度,并促使细胞周期通过G₀/G₁期,进入S期和G₂/M期。     

BARD1剪切变异体在卵巢癌组织中的表达及临床意义

目的探讨卵巢癌肿瘤抗原新基因OCY-142即BARD1的剪切变异体在卵巢癌中的表达及临床意义。方法用RT—PCR的方法检测OCY-142在卵巢癌组织和正常卵巢组织中的表达。结果在28例卵巢癌组织中,有10例（35．7％）明显表达OCY-142抗原基因,而30例正常卵巢组织中均没有OCY-142抗原基因的表达,两者之间有显著性差异（P〈0．05）。OCY-142的表达与卵巢癌的病理类型有关,在浆液性乳头状囊腺癌患者中有较高的特异性（P〈0．05）,但OCY-142的表达与卵巢癌的临床分期和组织分化程度没有相关性（P〉0．05）。结论卵巢癌肿瘤抗原基因OCY-142是BARD1基因的1个剪切变异体,其有可能成为卵巢恶性肿瘤免疫治疗的新靶点。OCY-142在卵巢浆液性囊腺癌中有较高的表达,其临床意义值得进行更深入的研究。     

Expression of drs mRNA in human lung adenocarcinomas

The drs gene was originally isolated from a rat primary embryo fibroblast cDNA library as a suppressor gene against v-src transformation. We have previously shown that expression of drs mRNA was markedly reduced in a variety of human cancer cell lines, including those of the colon, bladder, and ovary. Furthermore, introduction of drs cDNA by retrovirus vector into these cancer cell lines caused suppression of anchorage-independent growth without affecting cell proliferation. These findings suggest that down-regulation of drs mRNA is closely correlated with expression of malignant phenotypes in development of human cancers. To clarify the correlation between down-regulation of drs mRNA and malignant tumor formation in human tumor tissues, we examined the expression of drs mRNA in well-differentiated, moderately differentiated, and poorly differentiated lung adenocarcinoma tissues by in situ mRNA hybridization. The results clearly indicated that expression of drs mRNA was markedly reduced in 5 of 5 poorly differentiated lung adenocarcinomas examined but significantly expressed in normal lung tissues and 5 of 7 moderately-differentiated and 3 of 5 well-differentiated lung adenocarcinoma tissues. Neither gross deletion nor rearrangement of the drs genome was detected in these tissues. Down-regulation of drs mRNA was also observed in human lung adenocarcinoma cell lines derived from poorly differentiated adenocarcinomas. Our results suggest that down-regulation of drs mRNA is correlated with a poor degree of differentiation and progression of lung adenocarcinoma.     

Loss of Retinoic Acid Receptor-β Expression Is an Early Event during Esophageal Carcinogenesis

We recently observed that growth inhibition of esophageal cancer cells by retinoic acid (RA) was associated with both constitutive expression and RA-induced up-regulation of RA receptor beta (RAR-beta). Cell lines that did not express RAR-beta were also resistant to RA. To explore the expression of RAR-beta mRNA in vivo, we analyzed esophageal tissue specimens from 16 normal mucosae, 30 dysplastic lesions, and 157 esophageal tumors by in situ hybridization. RAR-beta was detected in 88% (14/16) of normal esophageal tissues and in 96% (96/100) of distant normal esophageal mucosa from cancer specimens. In contrast, RAR-beta was expressed in only 57% (17/30) of dysplastic lesions and in 54% (84/157) of carcinomas. Among esophageal carcinomas RAR-beta mRNA was expressed in 62% (26/42) of well-differentiated, 54% (27/50) of moderately differentiated, and only 29% (4/14) of poorly differentiated SCCs. Our data suggest that the loss of RAR-beta expression is an early event associated with esophageal carcinogenesis and the status of squamous differentiation.     

人肝细胞癌金属蛋白酶组织抑制因子-3的表达及与其基因启动子甲基化的关系

目的 探讨肝细胞癌的发生和门脉浸润机制。方法 20例肝细胞癌患者,每例在手术后分别取原发瘤、门脉瘤栓及远离肝癌之肝组织。用Western印迹法检测金属蛋白酶组织抑制因子(TIMP)的蛋白表达,用逆转录-聚合酶链反应(RT-PCR)检测TIMP-3 mRNA的表达,用甲基化特异性PCR检测TIMP-3基因启动子的甲基化。结果 远离肝癌之肝组织中均可见TIMP-3蛋白和mRNA的表达,原发瘤和门脉瘤栓组织TIMP-3蛋白和mRNA表达明显降低,其中分别有5例和6例完全丢失。远离肝癌之肝组织均未发现TIMP-3启动子甲基化。原发瘤中有7例、门脉瘤栓组织中有9例出现甲基化。所有有甲基化的肝癌组织,包括原发瘤和门脉瘤栓,有13例TIMP-3 mRNA和蛋白表达完全丢失,6例表达降低。TIMP-3启动子甲基化和肝细胞癌组织学分级无关(P>0.05)。结论 肝细胞癌的发生和门脉浸润与TIMP-3基因和蛋白缺失或降低相关、而TIMP-3启动子甲基化是其基因和蛋白缺失或降低的原因。     

肝细胞癌中ZHX2、NF-YA及AFP表达的相关关系

目的 检测肝细胞癌(hepatocellular carcinoma,HCC)组织中ZHX2、NF-YA及AFP mRNA和蛋白表达的相关关系.方法 采用RT-PCR及免疫组织化学方法检测HCC组织中ZHX2、NF-YA及AFP mRNA和蛋白的表达情况.结果 ZHX2 mRNA在AFP mRNA阴性的25例HCC组织中的表达率为52.0%,明显高于AFP mRNA阳性组织中的表达率(26.3%,P=0.038),ZHX2 mRNA与AFP mRNA表达呈负相关(OR=0.33);NF-YA mRNA与ZHX2 mRNA表达呈正相关(OR=31.429),而与AFP mRNA表达呈负相关关系(OR=0.308).AFP蛋白阴性的142例HCC组织中,ZHX2蛋白的表达率为23.9%,明显高于在AFP蛋白阳性组织中的表达率(12.8%,P=0.034);HCC组织中ZHX2蛋白与AFP蛋白表达呈负相关(P=0.018,r=-0.153).NF-YA蛋白在HCC组织中与ZHX2蛋白表达呈正相关(P=0.000,r=0.371),与AFP蛋白表达呈负相关(P=0.000,r=-0.497).结论 联合检测AFP和ZHX2蛋白将提高HCC的诊断率;ZHX2可能是通过NF-YA来调控AFP的表达.