c—erbB—2蛋白和表皮生长因子受体在肝脏病变中的表达

对１８４例乙型肝炎、肝硬变和肝细胞癌及２９例正常肝组织的标本作了ＡＢＣ法染色，观察基ｃ－ｅｒｂＢ－２蛋白和表皮生长因子受体的表达情况。３６％（４８／１３４）的慢性肝炎、肝硬变和ＨＣＣ组织中有ＥＧＦＲ表达，主要定位于血窦内皮。良、恶性病变肝组织之间在ＥＧＦＲ表达强度上无显著差别。提示ＥＧＦＲ可能与慢性肝脏闰变中血窦内皮的增生有关。     

人肝癌及肝硬变中IGF－2及HBxAg表达的对比研究

对６０例肝细胞癌、癌旁肝组织和４７例肝硬变石蜡切片进行ＩＧＦ－２和ＨＢｘＡｇ免疫组化染色。结果表明，在ＨＢｘＡｇ阳性的肝癌和肝硬变中ＩＧＦ－２阳性率分别为１００％（３２／３２例）和９４．６％（３５／３７例），而ＨＢｘＡｇ阴性组分别为８２．１％（２３／２８例）和６０％（６／１０例）。ＩＧＦ－２在肝癌中的阳性强度明显高于癌旁肝和肝硬变。ＩＧＦ－２的分布形态有：（１）胞浆包涵体样、（２）全胞浆、（３）核内分布三种。癌旁肝及肝硬变中可见小多角细胞（ＳＰＬＣ）呈ＩＧＦ－２和ＨＢｘＡｇ阳性。对ＩＧＦ－２在肝癌、癌旁肝和肝硬变中表达的意义及其与ＨＢｘＡｇ的关系以及ＳＰＬＣ的性质进行了讨论。     

EGR-1、c-erbB-2、ER、PR在乳腺癌中的表达及其相互关系

目的：探讨早期生长反应基因（ＥＧＲ－１）在乳腺癌的表达及其与癌基因ｃ－ｅｒｂＢ－２和ＥＲ、ＰＲ的关系。方法：应用免疫组织化学ＳＡＢＣ法,以ＥＧＲ－１、ｃ－ｅｒｂＢ－２、ＥＲ、ＰＲ抗体标记７５例不同病变的乳腺组织。结果：乳奶性病变可见相对高水平的ＥＧＲ－１表达而乳腺癌ＥＧＲ－１阳性仅１９例（４２．２％）,ＥＧＲ－１表达与乳腺癌淋巴结转移和ＥＲ、ＰＲ表达有关（Ｐ〈０．０５,Ｐ〈０．０１）,与ｃ－ｅｒｂ     

血小板衍生生长因子及其受体与病毒性肝炎肝纤维化的关系

目的研究血小板衍生生长因子在病毒性肝炎肝纤维化中的作用。方法以免疫组化技术检测血小板衍生生长因子（ＰＤＧＦ）－ＢＢ以及ＰＤＧＦ的β受体（ＰＤＧＦＲ－β）在病毒性肝炎肝组织中的表达。结果ＰＤＧＦ－ＢＢ及ＰＤＧＦＲ－β在肝组织中的表达与肝纤维化程度明显相关，肝硬变及慢性肝炎Ｓ３～４期患者肝组织ＰＤＧＦ－ＢＢ及ＰＤＧＦＲ－β表达强度明显高于急性肝炎及慢性肝炎Ｓ０～２期患者，同时与结蛋白、Ⅲ型前胶原肽在肝组织中的表达以及血清金属蛋白酶组织抑制因子－１（ＴＩＭＰ－１）呈明显正相关。结论ＰＤＧＦ－ＢＢ及ＰＤＧＦＲ－β不仅对星状细胞的活化、分裂、增殖及其合成胶原等细胞外基质（ＥＣＭ）均有明显的促进作用，而且可能通过上调ＴＩＭＰ－１减少ＥＣＭ的降解，促进肝纤维化的发生和进展过程。     

结直肠癌c—erbB—2,EGFR表达与肿瘤生物学行为的关系

目的：探讨结直肠癌ｃ－ｅｒｂＢ－２及表皮生长因子受体（ＥＧＦＲ）共同表达对其生物学行为及预后的影响。方法：应用免疫组织化学方法检测６９例结直肠癌中ｃ－ｅｒｂＢ－２及ＥＧＦＲ的表达情况。结果：ｃ－ｅｒｂＢ－２和ＥＧＦＲ共同表达时，肿癌细胞增殖指数高，呈浸润性生长方式的较多，病人５年生存率较低。结论：ｃ－ｅｒｂＢ－２和ＥＧＦＲ可能有协同作用，同时检验两者的表达情况可能为临床判断病人预后提供较可靠的依据     

c-erbB-2基因工程抗体的构建及在哺乳动物细胞中的表达

原癌基因ＨＥＲ－２／ｎｅｕ／ｅｒｂＢ－２编码的１８５－ｋＤａ跨膜磷酸糖蛋白为受体酪氨酸激酶家族中的成员。已报道ｃ－ｅｒｂＢ－２在多种腺癌中过表达，其中包括１５％～４０％的早期及转移性乳腺癌和卵巢癌［１］。亦有人报道，约７３％的转移性乳腺癌和卵巢癌过表达 ＨＥＲ－２／ｎｅｕ［２］，表明其为乳腺癌靶向性免疫治疗的理想靶分子。我们采用基因工程技术，构建了ｃ－ｅｒｂＢ－２ ＳｃＦｖ基因，并获得了其在ＣＯＳ７细胞中的表达。ｌ 材料和方法１．ｌ材料ＣＯＳ７细胞为本室保存。载体ｐｃＤＮＡ３．ｌ购自Ｉｎ－ｖｉｔｒ…     

p53、c-erbB-2、PCNA和EGFR在膀胱癌中过表达及其意义

目的：研究癌基因和抑癌基因蛋白产物在膀胱移行细胞癌中异常表达与病理分级、临床分期、复发和预后的关系。方法：应用免疫组化Ｓ－Ｐ法检查１１７例膀胱移行细胞癌组织中ｐ５３、ｃ－ｅｒｂＢ－２、ＰＣＮＡ和ＥＧＦＲ的表达水平。结果：１１７例膀胱移行细胞癌中ｐ５３、ｃ－ｅｒｂＢ－２、ＰＣＮＡ和ＥＧＦＲ阳性表达率分别为４７．０％、２９．９％、５３．８％和４８．７％。ｐ５３和ＰＣＮＡ阳性表达产物定位于肿瘤细胞核内，ｃ－ｅｒｂＢ－２阳性表达产物定位于细胞膜上，ＥＧＦＲ阳性表达产物定位于细胞膜或细胞浆内。结果表明ｐ５３、ｃ－ｅｒｂＢ－２、ＰＣＮＡ和ＥＧＦＲ异常表达与膀胱癌的分级、分期、复发及术后生存率等之间有统计学意义。结论：ｐ５３、ｃ－ｅｒｂＢ－２、ＰＣＮＡ和ＥＧＦＲ异常表达有助于评估膀胱癌预后，多基因异常表达作为预后评价指标更有意义。     

大肠癌c—erbB—2,EGFR及p21^ras蛋白共同表达与肿瘤细胞增殖

目的：搪塞ｃ－ｅｒｂＢ－２、ＥＧＦＲ及Ｐ２１＾ｒａｓ癌基因蛋白共同表达对大脾性癌细胞增殖的影响。方法：应用免疫组化ＡＢＣ法检测ｃ－ｅｒｂＢ－２、ＥＧＦＲｅｙ ｐ２１＾ｒａｓ蛋白在６９例大肠癌中的表达情况,同时计数肿瘤细胞ＰＣＮＡ增殖指数。结果：３种癌基因蛋白全部阳性,ｃ－ｅｒｂＢ＿２和ＥＧＦＲ同时阳性及单独ｃ－ｅｒｂＢ－２阳性的大肠癌,癌细胞ＰＣＮＡ指数高于３种蛋白全部阴性组。结果：ｃ－ｅｒｂＢ－     

血小板衍生生长因子及其受体与病毒性肝炎肝纤维化 …

目的 研究血小板衍生生长因子在病毒性肝炎肝纤维化中的作用。方法 以免疫组化技术检测血小板衍生生长因子（ＰＤＧＦ）－ＢＢ以及ＰＤＧＦ的β受体（ＰＤＧＦＲ－β）在病毒性肝炎肝组织中的表达。结果 ＰＤＧＦ－ＢＢ及ＰＤＧＦＲ－β在肝组织中的表达与肝纤维化程度明显相关，肝硬变及慢性肝炎Ｓ３￣４期患者肝组织ＰＤＧＦ－ＢＢ及ＰＤＧＦＲ－β表达强度明显高于急性肝炎及慢性肝炎Ｓ０￣２期患者，同时与结蛋白、Ⅲ型前胶原     

肝细胞提取物对肝癌细胞（BEL—7402）凋亡相关基因表达的影响

肝细胞提取物经ＨＰＬＣ，ＦＰＬＣ进一步分离纯化，得到了单一色谱峰，激光飞行时间质谱仪测分子量为４．０２０ｋＤ该单一组分（Ｓ４）对ＢＥＬ－７４０２肝癌细胞有增殖回落作用，半数抑制浓度（ＩＣ５０）为７．５８ｍｇ／Ｌ。对ＢＥＬ－７４０２细胞的基因表达影响研究，Ｓ４对Ｐ５３，Ｆａｓ表达有增强作用，而抑制Ｂｃｌ－２，ｃ－ｍｙｃ的表达，Ｓｏｕｔｈｅｒｍｂｌｏｔ技术表明，Ｓ４可抑制Ｂｃｌ－２的ｍＲＮＡ转录，提示     

Impact of epidermal growth factor receptor and transforming growth factor–α on hepatitis C virus‐induced hepatocarcinogenesis

Epidermal growth factor receptor system plays a central hepato‐protective and pro‐regenerative role in liver. Transforming growth factor‐α (TGF‐α) is an important autocrine growth regulator of hepatocytes that plays a role in development of hepatocellular carcinoma (HCC) among patients with chronic hepatitis C (CHC). This study was done on 40 core liver biopsies from patients with CHC, 20 liver specimens from HCC cases on top of CHC as well as five normal controls. All were immunohistochemically stained with epidermal growth factor receptor (EGFR) and TGF‐α antibodies. Some selected HCC cases were submitted for FISH technique to detect EGFR gene alteration. By immunohistochemistry EGFR and TGF‐α were overexpressed in HCC and cirrhotic cases compared to CHC cases without cirrhosis. Also, their expression was stronger in CHC cases with higher grades of activity and stages of fibrosis compared to lower ones. FISH positive results for EGFR were detected in 33.3% of the examined HCC cases. EGFR and TGF‐α can be used as predictive markers for activity, fibrosis, and carcinogenesis in CHC patients. Overexpression of EGFR in HCC patients can be promising in selecting those who can get benefit from anti‐EGFR target therapy.     

Liver cell dysplasia and hepatocellular carcinoma in non-A, non-B hepatitis

Liver cell dysplasia (LCD) is a premalignant cytologic change of hepatocytes that has been statistically linked to cirrhosis, hepatocellular carcinoma (HCC), and chronic liver disease related to hepatitis B virus. The relationship of LCD to non-A, non-B (NANB) hepatitis is currently unknown. We studied liver biopsy and surgical resection specimens from 36 patients with NANB hepatitis, and identified LCD in 17 (42.5%) of 40 specimens, most often associated with cirrhosis. Dysplasia was present in individual hepatocytes, in clusters, and in a distinctive "spreading" pattern of hepatocytes about central veins. Three patients had HCC with a predominant giant cell pattern, as well as LCD. These findings suggest that LCD and HCC should be included among the potential pathologic sequelae of NANB hepatitis.     

Immunohistochemical expression of CD95 (Fas), c-myc and epidermal growth factor receptor in hepatitis C virus infection, cirrhotic liver disease and hepatocellular carcinoma

Gene product expression in normal and chronic hepatitis C virus infection was determined in an attempt to improve our understanding of the molecular events leading to the development of cirrhosis and liver carcinoma. Activation of CD95 (Fas) causes apoptosis of cells and liver failure in mice and has been associated with human liver disorders. c-myc is involved in cell proliferation and EGFR in regeneration of cells. The material of the current study included 50 cases of chronic hepatitis C (CHC) (and negative hepatitis B virus infection), 29 cases of liver cirrhosis and HCV (LC), and 19 cases of hepatocellular carcinoma and HCV (HCC) admitted to the Theodor Bilharz Research Institute (TBRI) during the years 2003-2004. Ten wedge liver biopsies - taken during laparoscopic cholecystectomy - were included in the study as normal controls. Laboratory investigations, including liver function tests, serological markers for viral hepatitis and serum alpha fetoprotein level (alpha-FP), were determined for all cases. Histopathological study and immunohistochemistry using monoclonal antibodies for CD95, c-myc and EGFR were also done. In CHC cases, the histological activity index (HAI) revealed more expression of Fas antigen in liver tissues with active inflammation than in those without active inflammation (p < 0.01). EGFR and c-myc act synergistically in liver tumorigenesis. Upregulation of Fas in chronic hepatitis C infection and of c-myc & EGFR in malignant transformation was concluded from this study. c-myc expression may obstruct the induction of apoptosis of HCC cells and lead to uncontrolled cell growth.     

Analysis of proliferating hepatocytes using a monoclonal antibody against proliferating cell nuclear antigen/cyclin in embedded tissues from various liver diseases fixed in formaldehyde.

The authors studied histochemically the morphologic features of proliferating hepatocytes positive for proliferating cell nuclear antigen (PCNA/cyclin) to analyze the process of liver regeneration in embedded tissues fixed with formaldehyde using an anti-PCNA/cyclin monoclonal antibody. In liver specimens from patients with acute viral hepatitis (AVH) and confluent necrosis, many small basophilic hepatocytes surrounding large clear hepatocytes were positively stained in the areas next to the confluent necrosis. Therefore these small hepatocytes may be daughter cells derived from large clear hepatocytes that probably enter the mitotic cell cycle repeatedly to repair a large necrotic area. In the case of AVH with spotty necrosis, the positively stained hepatocytes were scattered around the necrotic foci. In the liver specimens from patients with chronic active hepatitis, most of the positively stained hepatocytes were located next to the necrotic area. As for cirrhosis of the liver, the number of hepatocytes positive for PCNA/cyclin varied greatly in different pseudolobules, and in the specimens of hepatocellular carcinoma (HCC), the HCC cells positive for PCNA/cyclin were detected throughout the cancer nests.     

Immunolocalization of alpha interferon in liver disease.

The expression of immunoreactive alpha interferon was examined in 78 liver biopsy specimens using an indirect immunoperoxidase technique. Biopsy specimens included cases of acute viral hepatitis, chronic active hepatitis, primary biliary cirrhosis, alcoholic hepatitis, large bile duct obstruction and normal liver. Kupffer cells were positive for alpha interferon in all cases. Hepatocytes were negative for alpha interferon in normal liver but in acute viral hepatitis were positive in perivenular and necrotic areas. Hepatocytes were positive in periportal areas, associated with piecemeal necrosis, in chronic active hepatitis and primary biliary cirrhosis, and were positive in perivenular areas in alcoholic hepatitis and large bile duct obstruction. The unexpected finding of alpha interferon in hepatocytes in non-viral liver disease indicates that the presence of this substance in liver cells cannot be taken as a specific marker of viral infection.     

Expression of TLR-2 in hepatocellular carcinoma is associated with tumour proliferation,angiogenesis and Caspase-3 expression

AimsUnlike other Toll-like receptors (TLRs), the role of toll like receptor 2 (TLR-2) in the pathogenesis of chronic liver disease and hepatocellular carcinoma (HCC) is not well studied. We, therefore, set out to investigate the expression of TLR-2 in different chronic liver disease states along with other markers of cell death, cellular proliferation and tissue vascularisationMethods and resultsImmunohistochemistry was performed on liver tissue microarrays comprising hepatitis, cirrhosis and HCC patient samples using antibodies against TLR-2, Ki-67, Caspase-3 and VEGF. This was done in order to characterise receptor expression and translocation, apoptosis, cell proliferation and vascularisation. Cytoplasmic TLR-2 expression was found to be weak in 5/8 normal liver cases, 10/19 hepatitis cases and 8/21 cirrhosis patients. Moderate to strong TLR-2 expression was observed in some cases of hepatitis and cirrhosis. Both, nuclear and cytoplasmic TLR-2 expression was present in HCC with weak intensity in 11/41 cases, and moderate to strong staining in 19/41 cases. Eleven HCC cases were TLR-2 negative. Surprisingly, both cytoplasmic and nuclear TLR-2 expression in HCC were found to significantly correlate with proliferative index (r = 0.24 and 0.37), Caspase-3 expression (r = 0.27 and 0.38) and vascularisation (r = 0.56 and 0.23). Further, nuclear TLR-2 localisation was predominant in HCC, whereas cytoplasmic expression was more prevalent in hepatitis and cirrhosis. Functionally, treatment of HUH7 HCC cells with a TLR-2 agonist induced the expression of cellular proliferation and vascularisation markers CD34 and VEGF.ConclusionsOur results demonstrate a positive correlation between the expression of TLR-2 and other markers of proliferation and vascularisation in HCC which suggests a possible role for TLR-2 in HCC pathogenesis     

An adiponectin receptor, T-cadherin, was selectively expressed in intratumoral capillary endothelial cells in hepatocellular carcinoma: possible cross talk between T-cadherin and FGF-2 pathways

T-cadherin is a unique receptor of adiponectin, which plays a critical role in various angiogenesis. In the present study, T-cadherin expression in tumor vessels of hepatocellular carcinoma (HCC) and, subsequently, the molecular mechanism, which induced T-cadherin expression in sinusoidal endothelial cells were investigated. Sinusoidal endothelium in nontumorous liver, chronic hepatitis, or liver cirrhosis expressed little or no T-cadherin. By contrast, T-cadherin was found in intratumoral capillary endothelial cells of 34 out of 63 HCC specimens. In positive cases, focal T-cadherin expression was found in well-differentiated HCC, whereas diffuse and intense T-cadherin expression was observed in poorly differentiated HCC specimens. T-cadherin was much expressed in intratumoral capillary endothelial cells in a less differentiated HCC region than that in a well-differentiated region in five specimens, in which various differentiated HCC components were coexistent. In a double-cell chamber assay, fibroblast growth factor-2 appeared to have a critical role to induce T-cadherin in cultured liver sinusoidal endothelial cells. The present finding indicated that T-cadherin was selectively expressed in intratumoral capillary endothelial cells of many HCCs, increasingly expressed as tumor progression, and T-cadherin may have a positive role in angiogenesis of HCC. In addition, cross talk between the signal pathways mediated by fibroblast growth factor-2 and adiponectin was suggested.