Raf kinase inhibitor protein is downregulated in hepatocellular carcinoma

The Ras/Raf/MEK/ERK signalling cascade is frequently deregulated in tumourigenic diseases and known to be involved in proliferation and transformation of cells. Also in hepatocellular carcinoma (HCC) increased ERK levels are observed and known to correlate with tumour progression, but the underlying molecular mechanism are unknown. We analyzed expression of Raf-1 kinase inhibitory protein (RKIP) in HCC. Expression of RKIP mRNA and protein was downregulated in HCC cell lines and tissue as compared to primary human hepatocytes (PHH) or non-tumorous liver tissue, respectively. Transfection of an HCC cell line with an RKIP expression construct blocked the Raf kinase pathway resulting in decreased activity of ERK1/2 and AP-1. In contrast, downregulation of RKIP by transfection with an antisense RKIP construct led to increased ERK1/2 and AP-1 activity. Since HCC develop in the majority of cases in cirrhotic liver tissue and cirrhosis is the main risk factor for HCC development, we analyzed RKIP expression also in non-cancerous cirrhotic liver tissues by immunohistochemistry. In contrast to normal liver tissue, where the staining was equally distributed within the cytoplasm, hepatocytes in cirrhotic liver revealed an intense RKIP staining of the membrane. It can be speculated that this changed RKIP expression pattern parallels impaired protein function in PHH in cirrhotic livers that may predispose PHH to malignant transformation. In addition, our study demonstrates functional relevance of downregulation of RKIP in HCC that may play an important role in HCC development and progression.     

Tricellulin Expression and its Prognostic Significance in Primary Liver Carcinomas

Numerous data suggest that altered expression of tight junction proteins such as occludin and claudins plays important role in carcinogenesis. However, little is known about tricellulin, a transmembrane tight junction protein concentrated where three epithelial cells meet. We aimed to characterize tricellulin expression in normal and cirrhotic liver in comparison to primary hepatic neoplasms. Tricellulin expression of 20 control livers, 12 cirrhotic livers, 32 hepatocellular carcinomas (HCC), and 20 intrahepatic cholangiocarcinomas (iCCC) was investigated by immunohistochemistry and Western blotting. Co-localization of tricellulin with claudin-1, -4, and MRP2 was studied using double immunofluorescence. Scattered tricellulin immunopositivity was restricted to biliary pole of hepatocytes confirmed by co-localization with MRP2. Moreover, spotted-like reaction was observed between bile duct epithelial cells. In 40 % of HCCs marked tricellulin overexpression was measured regardless of tumor grades. In iCCCs, however, tricellulin expression decreased parallel with dedifferentiation. In HCCs high tricellulin expression, in iCCCs low tricellulin expression correlated with poor prognosis. Co-localization with MRP2 might substantiate that tricellulin plays role in blood-biliary barrier. Overexpressed tricellulin in a subset of HCCs correlated with unfavorable prognosis. Similar to ductal pancreatic adenocarcinoma, higher grades of iCCCs were associated with decreased tricellulin expression correlating with poor prognosis.     

Comparison study of the expressions of myristoylated alanine-rich C kinase substrate in hepatocellular carcinoma, liver cirrhosis, chronic hepatitis, and normal liver

The myristoylated alanine-rich C kinase substrate (MARCKS) is a prominent substrate for protein kinase C (PKC) in a variety of cells. The aim of this study was not only to evaluate the expression and localization of MARCKS in various pathological liver tissues, including HCC, but also to analyze the difference in MARCKS expression between hepatitis virus-induced HCC and cirrhosis. The level of MARCKS and its phosphorylated proteins, as well as its localization, were determined using Western blot and/or immunohistochemistry in HCC and other pathological liver tissues. We also analyzed the change of MARCKS localization on the influence of MARCKS phosphorylation in the HLF cancer cell line by phosphorylation study. In addition, the relationship between MARCKS expression and proliferative activity was studied in HCC. In the immunohistochemical study, a very small amount of MARCKS protein was found along the contour of the hepatocellular membrane in normal liver and in cases of chronic hepatitis. MARCKS was up-regulated in liver cirrhosis tissue and was localized in the cytoplasm of hepatocytes. The expression of MARCKS was down-regulated in HCC tissues, as compared with non-tumorous liver cirrhosis tissues from the same patients. Furthermore, MARCKS was serine-phosphorylated in liver cirrhosis and HCC, and phosphorylated MARCKS was detected in a cytosolic fraction of these tissues. In a phosphorylation study using the HLF HCC cell line, MARCKS was displaced from the plasma membrane to the cytosol following the activation of protein kinase C (PKC) by phorbol 12-myristrate 13-acetate (PMA). Furthermore, the activity of cyclin D1 and cyclin E kinases was found to be higher in HCCs with low MARCKS expression than in HCCs with high MARCKS expression. These results suggest that up-regulation of MARCKS might be essential in the generation of cirrhotic nodules through chronic hepatitis from normal liver, and that the phosphorylation and/or down-regulation of MARCKS might play an important role in the development and progression of HCC from liver cirrhosis.     

Activation of a system A amino acid transporter, ATA1/SLC38A1, in human hepatocellular carcinoma and preneoplastic liver tissues

The expression of amino acid transporter (AT) mRNAs including A system (ATA1/SNAT1/SLC38A1, ATA2/SNAT2/SLC38A2 and ATA3/SNAT3/SLC38A4), L system (LAT1/SLC7A5 and LAT2/SLC7A8), and y+ (CAT2/SLC7A2) genes, were compared among hepatocellular carcinoma (HCC) and non-cancerous liver cells. Among them the ATA1 mRNA expression was significantly elevated in all HCC cell lines (HepG2, HLF, HuH7 and JHH4) examined compared with normal liver tissue. We further discovered that the expression of ATA1 mRNA was significantly activated in HCC tissues and also elevated in pre-malignant cirrhotic livers from HCC patients, compared with normal livers from non-HCC patients. The ATA1 protein was extensively accumulated in the cytoplasm of pre-malignant liver and most HCCs, while being weak or undetectably low in normal liver tissues. SiRNA-mediated suppression of endogenous ATA1 lowered the viability of HepG2 cells. Thus, the activation of ATA1 confers growth and survival advantages in pre-malignant and malignant liver lesions.     

紧密连接相关蛋白在胃腺癌中的联合表达及预后意义

  目的  探讨紧密连接蛋白(Claudin-1、-3、-4和-7)在胃腺癌纵向侵袭转移中的表达变化及预后意义。  方法  应用组织芯片和免疫组化SP法检测139例胃腺癌患者黏膜腺上皮(含肠化生上皮), 黏膜层, 肿瘤中心, 侵袭前沿及淋巴结转移灶胃癌组织Claudin-1、-3、-4、-7的表达水平, 分析其与胃腺癌侵袭转移及预后关系。  结果  Claudin-1、-3、-4和-7在肠化生上皮的总体表达水平高于胃黏膜腺上皮(P < 0.05);从黏膜层→肿瘤中心→侵袭前沿→淋巴结转移灶, 胃癌组织Claudin-1、-3、-4和-7的表达呈抛物线样趋势, 肿瘤中心表达最高(χ2=111.84, 52.35, 33.71和111.91, P < 0.05);肿瘤中心Claudin-1和-3表达降低, 患者预后不良(χ2=5.530和χ2=9.846, P < 0.05), Cox多因素生存分析发现仅Claudin-3和临床分期可作为胃癌患者的预后独立预测因素。  结论  Claudin-1、-3、-4和-7在胃癌纵向侵袭转移中的表达变化可能与胃黏膜的肠上皮化生、肿瘤增殖及侵袭转移中的上皮-间质转化有关。      

Patterns of expression of cytochrome P450 genes in progression of hepatitis C virus-associated hepatocellular carcinoma

Cytochrome P450 (CYP) genes are involved in the pathogenesis of hepatocellular carcinoma (HCC). To examine changes in expression of CYPs in HCC arising from hepatitis C virus (HCV)-infected liver, we used oligonucleotide array data of 27 CYPs from samples of 50 HCV-associated HCCs, five HCV-infected non-tumorous livers, and six HCV-negative normal livers. Progression of primary HCC can be characterized by decrease in the grade of tumor differentiation, increased frequency of venous invasion and increased tumor size. On the basis of tumor differentiation, the self-organizing map (SOM) classified the 27 CYPs into four groups. The first group contained 11 CYPs, including the CYP2C and CYP4F families, that showed decreased expression in parallel with progression of HCV-infected liver to HCC with less differentiation. The second group contained CYP-IID, CYP3A7 and CYP27A1, genes that showed high levels of expression specific to well differentiated HCC. The third group contained 5 sterol-metabolizing CYPs with levels lower in HCV-infected livers than in HCV-uninfected livers. The last group included the CYP2E1 and CYP3A families. Among the 27 CYPs, levels of 7 (CYP2B6, CYP-IIC, CYP2C9, CYP2C19, CYP3A5, CYP4F3 and CYP27A1) were significantly lower and levels of 2 (CYP2E1 and CYP4F2) were slightly lower in HCC with venous invasion than in HCC without venous invasion. Levels of CYP-IIC and CYP2C9 were inversely associated with tumor size. In contrast, levels of CYP51A1 were positively associated with tumor size. Our present study revealed that expression of specific CYPs was altered in conjunction with progression of HCV-associated HCC. These CYPs may serve as markers of progression and molecular targets for treatment of HCV-associated HCC.     

Claudin-2 expression increases tumorigenicity of colon cancer cells: role of epidermal growth factor receptor activation

Claudin-2 is a unique member of the claudin family of transmembrane proteins, as its expression is restricted to the leaky epithelium in vivo and correlates with epithelial leakiness in vitro. However, recent evidence suggests potential functions of claudin-2 that are relevant to neoplastic transformation and growth. In accordance, here we report, on the basis of analysis of mRNA and protein expression using a total of 309 patient samples that claudin-2 expression is significantly increased in colorectal cancer and correlates with cancer progression. We also report similar increases in claudin-2 expression in inflammatory bowel disease-associated colorectal cancer. Most importantly, we demonstrate that the increased claudin-2 expression in colorectal cancer is causally associated with tumor growth as forced claudin-2 expression in colon cancer cells that do not express claudin-2 resulted in significant increases in cell proliferation, anchorage-independent growth and tumor growth in vivo. We further show that the colonic microenvironment regulates claudin-2 expression in a manner dependent on signaling through the EGF receptor (EGFR), a key regulator of colon tumorigenesis. In addition, claudin-2 expression is specifically decreased in the colon of waved-2 mice, naturally deficient in EGFR activation. Furthermore, genetic silencing of claudin-2 expression in Caco-2, a colon cancer cell line, prevents the EGF-induced increase in cell proliferation. Taken together, these results uncover a novel role for claudin-2 in promoting colon cancer, potentially via EGFR transactivation.     

Alteration of the copy number and deletion of mitochondrial DNA in human hepatocellular carcinoma

Somatic mutations in mitochondrial DNA (mtDNA) have been detected in hepatocellular carcinoma (HCC). However, it remains unclear whether mtDNA copy number and mitochondrial biogenesis are altered in HCC. In this study, we found that mtDNA copy number and the content of mitochondrial respiratory proteins were reduced in HCCs as compared with the corresponding non-tumorous livers. MtDNA copy number was significantly reduced in female HCC but not in male HCC. Expression of the peroxisome proliferator-activated receptor gamma coactivator-1 was significantly repressed in HCCs (P<0.005), while the expression of the mitochondrial single-strand DNA-binding protein was upregulated, indicating that the regulation of mitochondria biogenesis is disturbed in HCC. Moreover, 22% of HCCs carried a somatic mutation in the mtDNA D-loop region. The non-tumorous liver of the HCC patients with a long-term alcohol-drinking history contained reduced mtDNA copy number (P<0.05) and higher level of the 4977 bp-deleted mtDNA (P<0.05) as compared with non-alcohol patients. Our results suggest that reduced mtDNA copy number, impaired mitochondrial biogenesis and somatic mutations in mtDNA are important events during carcinogenesis of HCC, and the differential alterations in mtDNA of male and female HCC may contribute to the differences in the clinical manifestation between female and male HCC patients.     

Is mallory body formation a preneoplastic change? A study of 181 cases of liver bearing hepatocellular carcinoma and 82 cases of cirrhosis

The hypothesis that Mallory body formation by hepatocytes is a sign of preneoplasia was tested. This hypothesis was based on animal experiments but has not been tested in man. The authors studied the livers of 181 human autopsies in which hepatocellular carcinoma (HCC) was present and 82 cirrhotic livers from patients with alcoholism, HB viral infection, or cryptogenic cirrhosis. The frequency of Mallory bodies in nonneoplastic hepatocytes was 40% in the HCC-bearing livers with cirrhosis (LC). In HCC-bearing livers with pre-cirrhotic changes (PC), 25% showed Mallory body formation by nonneoplastic hepatocytes. In the cases of HCC, where there was no accompanying PC or LC, Mallory bodies were never found in the nonneoplastic hepatocytes. When the 82 cirrhotic livers without HCC and the 116 cirrhotic cases with HCC were combined, it was found that HCC was present in 70% of cirrhotic livers when the nonneoplastic liver cells contained Mallory bodies. When no Mallory bodies were found in the nonneoplastic liver cells, HCC was present in 53% of cases. The difference between the two groups was significant (P less than 0.05). The difference was significant for both HB viral hepatitis and cryptogenic cirrhosis but not for alcoholic cirrhosis. Likewise, when nonneoplastic hepatocytes formed Mallory bodies in cirrhotic livers, there was a statistically significant increase in the number of HCC cells that formed Mallory bodies (P less than 0.01). When nonneoplastic hepatocytes occurred in groups of Mallory body forming cells, the hepatocellular features were atypical and characteristic of dysplastic cells. The evidence indicates that when Mallory body formation was observed in HBsAg-positive and cryptogenic cirrhotic livers, they were associated with an increased frequency of HCC formation in man.     

Dominance of functional androgen receptor allele with longer CAG repeat in hepatitis B virus-related female hepatocarcinogenesis

The CAG polymorphism in exon 1 of the androgen receptor (AR) gene has been shown associated with the development of human male hepatocellular carcinoma (HCC) with the shorter AR alleles conferring a higher risk. However, the significance of AR-CAG repeats in female hepatocarcinogenesis remains to be addressed. In this study, seventy-six pairs of female HCCs and corresponding nontumorous tissues were collected, and 180 cirrhotic nodules were microdissected from 7 cirrhotic livers. The clonality status, functional AR alleles, and CAG repeat number of each sample were determined by AR methylation analysis. In a total of 44 monoclonal HCCs, the mean of CAG repeats in the active alleles was significantly longer than that in the inactive alleles (22.0 +/- 2.8 versus 20.7 +/- 3.6; P = 0.047). When we divided HCCs into hepatitis B virus-positive [HBV(+)] and HBV(-) subgroups, the long AR allele dominance was found only in HBV(+) ones (P = 0.006 versus P = 0.923). Notably, the preference of long CAG repeat has also been found in the 100 monoclonal nodules (P = 0.013). For comparison of monoclonal nodules obtained from the same individual, a dominant long AR allele was found in 6 patients. The proportion of monoclonal cirrhotic nodules and HCCs expressing longer AR allele, 69 and 68%, are both significantly higher than 50%, the assumed value in normal liver (P < 0.001 for cirrhotic nodules and P = 0.005 for HCC). The dominance is again only prominent in HBV-infected HCCs [85% for HBV(+) HCC; P < 0.001 but 54% for HBV(-) HCC; P = 0.27]. The results indicated that in female hepatocarcinogenesis, hepatocytes expressing the longer AR allele seem to be favorably selected for autonomous growth and transformation, especially in synergy with HBV infection.     

Augmented hepatocellular carcinoma progression and depressed Kupffer cell activity in rat cirrhotic livers

To examine the property of hepatocellular carcinoma (HCC), rat HCC cells were implanted into normal and cirrhotic rat livers. Implanted HCC grew much more progressively in cirrhotic livers than in normal livers. Kupffer cells were decreased profoundly in cirrhotic livers, resulting in markedly impaired phagocytic activity. Furthermore, production of Kupffer cell-related cytokines, such as interleukin-1 beta and tumor necrosis factor-alpha, was decreased profoundly in cirrhotic livers. Our results indicate that liver cirrhosis is a prominent promoting factor in HCC progression, and that markedly depressed Kupffer cell activity may play a role in augmented HCC progression in cirrhotic livers.     

TC21/R-Ras 2在肝癌中的表达、亚细胞定位及意义

目的:探讨TC21基因在肝癌细胞、肝癌及癌旁组织中的表达、亚细胞定位及意义。方法:用RT-PCR检测TC 21在肝癌细胞系及人肝癌及癌旁组织中mRNA水平的表达,用免疫组化及Western印迹检测TC21蛋白在肝癌相关组织中的表达差异,用组织芯片技术结合免疫组化检测TC21蛋白的亚细胞定位。结果:RT-PCR检测结果表明TC21 mRNA在SMMC-7721、BEL-7402、Huh-7、Hep3B、HepG2、MHCC-97L、MHCC-LM3 7个肝癌细胞系及L-02和Chang liver 2株永生化人肝细胞系、7对肝癌及对应癌旁肝组织中均有较高表达;Western印迹结果表明上述细胞系中TC21蛋白表达与mRNA表达一致,TC21在4对肝癌及癌旁组织中均呈较高水平的表达;免疫组化检测结果表明TC21在人原发性肝癌、癌旁组织、硬化肝组织与正常肝组织中的表达阳性率分别为84.2%、77.2%、41.7%、0%,肝癌与肝硬化、正常肝相比显著高表达(P<0.05,P<0.01),与癌旁组织比无统计学意义(P>0.05);统计学分析结果表明TC21蛋白表达与肿瘤大小呈正相关(P<0.05),与是否肝硬化呈负相关(P<0.05);肝癌组织中TC21主要定位于肝癌细胞核,部分定位于胞质,膜定位不明显,TC21蛋白在肝癌细胞中的核定位显著高于癌旁肝细胞(P<0.001)。结论:本研究首次揭示TC21蛋白在肝癌组织中的高表达及核定位预示其在肝细胞恶性转化及肝癌发生发展中发挥重要作用。     

Microsatellite instability mutator phenotype in hepatocellular carcinoma in non-alcoholic and non-virally infected normal livers

Microsatellite instability (MSI) seems to be a rare event in hepatocarcinogenesis and might actually be associated with the progression of hepatocellular carcinoma (HCC) in which the liver is often the site of chronic hepatitis or cirrhosis. The aim of this work was to define the MSI phenotype in HCC affecting exclusively normal livers to avoid slippage errors due to cirrhosis. One hundred and sixty-four patients with HCC affecting non-cirrhotic livers were operated on in our hospital between 1984 and 2001. We analyzed 37 patients selected for low alcohol consumption and the absence of HBV or HCV infection. All the livers were histologically normal. MSI was analyzed according to the criteria defined during the conference consensus workshop for colorectal cancer. High MSI (MSI-H > 30%) was found in 6 (16%) and low MSI (MSI-L < 30%) in 10 (27%) of the 37 HCCs. None of the 10 microsatellite markers tested were altered in the remaining 21 tumors (57%). Immunohistochemistry showed that normal amounts of hMLH1 and hMSH2 were present both in MSI-H and in MSI-L HCCs. MSI-H was significantly associated with more aggressive histological tumor features and a shorter median delay before recurrence. Thus, we have found a small subgroup of HCC tumors which can be considered as a new clinical/histological entity.     

Identification of genes preferentially methylated in hepatitis C virus‐related hepatocellular carcinoma

Chronic infections by hepatitis B virus (HBV) and hepatitis C virus (HCV) appear to be the most significant causes of hepatocellular carcinoma (HCC). Aberrant promoter methylation is known to be deeply involved in cancer, including in HCC. In this study, we analyzed aberrant promoter methylation by methylated DNA immunoprecipitation‐on‐chip analysis on a genome‐wide scale in six HCCs including three HBV‐related and three HCV‐related HCCs, six matched noncancerous liver tissues, and three normal liver tissues. Candidate genes with promoter methylation were detected more frequently in HCV‐related HCC. Candidate genes methylated preferentially to HBV‐related or HCV‐related HCCs were detected and selected, and methylation levels of the selected genes were validated by quantitative methylation analysis using MALDI‐TOF mass spectrometry using 125 liver tissue samples, including 61 HCCs (28 HBV‐related HCCs and 33 HCV‐related HCCs) and 59 matched noncancerous livers, and five normal livers. Among analyzed genes, preferential methylation in HBV‐related HCC was validated in one gene only. However, 15 genes were found to be methylated preferentially in HCV‐related HCC, which was independent from age. Hierarchical clustering of HCC using these genes stratified HCV‐related HCC as a cluster of frequently methylated samples. The 15 genes included genes inhibitory to cancer‐related signaling such as RAS/RAF/ERK and Wnt/β‐catenin pathways. Methylation of dual specificity phosphatase 4 (DUSP4), cytochrome P450, family 24, subfamily A, polypeptide 1 (CYP24A1), and natriuretic peptide receptor A (NPR1) significantly correlated with recurrence‐free survival. It was indicated that genes methylated preferentially in HCV‐related HCC exist, and that DNA methylation might play an important role in HCV‐related HCC by silencing cancer‐related pathway inhibitors, and might perhaps be useful as a prognostic marker. (Cancer Sci 2010)     

Hepatocellular carcinoma in an orthotopic mouse model metastasizes intrahepatically in cirrhotic but not in normal liver

Prognosis of hepatocellular carcinoma (HCC) still remains poor mainly because of intrahepatic metastasis. In the majority of cases, HCC is found in conjunction with liver cirrhosis. It is, therefore, of great importance to investigate the invasive and metastatic behavior of HCC in cirrhotic liver. To examine this, a liver cirrhosis model was produced by injecting thioacetamide i.p. into mice. Murine HCC cells were labeled with the fluorescent carbocyanine dye, DiI, and implanted directly under the capsule of cirrhotic and normal livers of syngeneic mice. DiI‐labeled HCC cells in the liver were observed under fluorescent and confocal microscopy. Histological analysis of cirrhotic and normal livers revealed that implanted HCC cells migrated to and invaded the adjacent periportal regions, but not the adjacent centrolobular areas. This characteristic behavior of HCC was more evident in cirrhotic liver than in normal liver. Furthermore, intrahepatic metastasis to unimplanted hepatic lobes was observed in cirrhotic liver as early as 7 days after implantation, while it was not detected in normal liver even 4 weeks later. Thus, an orthotopic animal model for HCC with cirrhosis described here may be suitable for investigating the invasive and metastatic behavior of HCC. Importantly, labeling tumor cells with a fluorescent dye before orthotopic implantation may be a convenient and useful method to investigate the invasive and metastatic behavior of various types of cancer. Int. J. Cancer 80:471–476, 1999. © 1999 Wiley‐Liss, Inc.     

Difference in histological activity of non-tumorous liver tissue between hepatitis B virus- and hepatitis C virus-associated hepatocellular carcinoma without cirrhosis

To elucidate the difference in the liver carcinogenetic process during hepatitis B virus (HBV) and hepatitis C virus (HCV) infections, non-tumorous liver tissues obtained from 10 patients who developed HBV-associated hepatocellular carcinoma (HCC) without cirrhosis were compared with those obtained from 26 patients who developed HCV-associated HCC without cirrhosis. The extent of fibrosis was similar in both groups. In contrast, necroinflammatory activities were significantly higher in patients with HCV than in patients with HBV. These results indicate that ongoing liver inflammation mediates the hepatocarcinogenesis more pronouncedly in HCV infection than in HBV infection.     

肿瘤-睾丸抗原SSX1和SSX4的mRNA在肝细胞肝癌中的表达

目的检测肿瘤-睾丸抗原(Cancer-Testis antigens,CT)SSX1和 SSX4基因 mRNA 在肝细胞肝癌(HCC)中的表达,探讨 CT 抗原 SSX1和 SSX4 mRNA 在 HCC 中表达有无特异性。方法应用逆转录-聚合酶链反应(RT-PCR)方法对35例 HCC 患者癌组织和相应的癌旁组织 SSX1和 SSX4基因表达进行检测,对其中6例 RT-PCR 扩增产物的目的片段进行 DNA 序列测定。结果 35例 HCC 患者中,SSX1和 SSX4的表达率分别为81%(27/35)和73%(23/35);癌旁组织、12例肝硬化和15例正常组织未检测到 SSX1和 SSX4的表达。6例 DNA 序列测定结果显示 RT-PCR 产物为 SSX1和 SSX4基因的 cDNA。SSX1和 SSX4的表达与患者年龄、性别、肿瘤大小、分化程度、血清甲胎蛋白(AFP)水平、乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)感染无显著相关性(P>0.05)。结论 SSX1和 SSX4在 HCC 中呈高特异性表达,是制备 HCC 疫苗理想的靶向分子,多种 CT 抗原联合表达为多效价疫苗治疗 HCC提供了理论基础。     

肿瘤-睾丸抗原SSX1和SSX4的mRNA在肝细胞肝癌中的表达

目的 检测肿瘤-睾丸抗原(Cancer-Testis antigens，CT)SSX1和SSX4基因mRNA在肝细胞肝癌(HCC)中的表达，探讨CT抗原SSX1 和SSX4 mRNA在HCC中表达有无特异性。 方法 应用逆转录-聚合酶链反应(RT-PCR)方法对35例HCC患者癌组织和相应的癌旁组织SSX1和SSX4基因表达进行检测，对 其中6例RT-PCR扩增产物的目的片段进行DNA序列测定。 结果 35例HCC患者中，SSX1和SSX4的表达率分别为81％(27／35)和73％(23／35)；癌旁组织、12例肝硬化和15例正常组织未 检测到SSX1和SSX4的表达。6例DNA序列测定结果显示RT-PCR产物为SSX1和SSX4基因的cDNA。SSX1和SSX4的表达与患 者年龄、性别、肿瘤大小、分化程度、血清甲胎蛋白(AFP)水平、乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)感染无显著相关性(P >0．05)。 结论 SSX1和SSX4在HCC中呈高特异性表达，是制备HCC疫苗理想的靶向分子，多种CT抗原联合表达为多效价疫苗治疗HCC 提供了理论基础。