细胞角质素CK18和CK19在肝细胞癌组织中的表达

目的:观察和探讨细胞角质素CK18和CK19在不同肝组织中的表达及其意义.方法:采用免疫组化SP法观察正常肝组织8例.肝硬化组织27例和肝细胞癌(hepatocellular carcinoma,HCC)组织43例中CK18,CK19的表达.结果:肝CK18和CK19在肝硬化组与正常肝组织的表达率均无明显差异,但两者在肝细胞癌组与肝硬化组之间差异有显著性(65.1%vs29.6%.69.8%vs 25.9%,P＜0.01).在20/27例肝硬化组织和35/43例HCC组织中可见到CK18,CK19标记的卵圆细胞,且在两种组织中卵圆细胞的数量有显著性差异(CK18:6.57-4-1.69vs 10.70±2.31:CK19:5.37±1.17vs10.45±2.15,P＜0.01).结论:CK18和CK19参与了肝硬化到HCC的癌变过程;CK18和CK19在卵圆细胞中的强阳性表达,支持HCC的卵圆细胞起源学说.     

肝细胞癌中DcR3表达与卵圆细胞增殖的可能关系

目的研究DcR3（一种新的凋亡相关分子）在肝细胞癌（HCC）中表达情况及其与卵圆细胞增殖之间可能存在的关系,以探讨卵圆细胞增殖活化的分子机制。方法对52例HCC组织,用免疫组织化学EnVision法同时标记肝细胞分化标记物hepatocyte paraffinl（HEP）、胆管标记物CK7/CK19、细胞增殖标记物Ki-67以及DcR3,半定量分析卵圆细胞形态数量和凋亡细胞数量的关系。结果在所有HCC肿瘤旁硬化组织的反应性小胆管中,均可见CK7、CK19和DcR3的表达,而在52例HCC中有23例见部分肿瘤细胞表达CK7和/或CK19,36例部分肿瘤细胞表达DcR3。其中23例CK7和/或CK19阳性病例中,21例同时有DcR3阳性（91.3%）,而29例CK7/CK19阴性病例中仅有15例（51.7%）DcR3阳性表达,两组间差异有统计学意义（P=0.002）。结论HCC中存在CK7和/或CK19阳性表达的卵圆细胞,表明卵圆细胞在部分HCC的发病中起重要作用,而这些病例多数有DcR3高表达,同时可见DcR3高表达于癌旁硬化组织反应性小胆管中,这些均提示DcR3的抗凋亡作用可能与卵圆细胞的增殖分化有关。     

Thy1.1阳性肝脏卵圆细胞在大鼠肝硬化形成与消减过程中的动态表达

目的 肝脏卵圆细胞（HOC）在二甲基亚硝胺（DMN）致大鼠肝硬化形成与消减过程中表达的动态变化，探讨其病理生理意义。方法 应用DMN腹腔注射（4周12次）制备大鼠肝硬化模型，分别于造模后3d、2、4周及终止造模后2、4周处死大鼠取肝组织，进行常规组织学观察，透射电镜下观察HOC超微结构，免疫组织化学和western blot方法检测Thy1．1的表达变化，应用图像分析法对所标记的HOC进行半定量测定以及光镜下计数细胞数量。结果 DMN造模4周大鼠已形成典型的肝硬化；终止造模后2周时肝组织病变较造模4周时有所减轻，终止造模后4周时炎症明显减轻并以不完全纤维间隔为主。Thy1．1阳性染色细胞正常大鼠肝组织未见到，造模后3d时可见微量表达，2周时呈散在分布，4周时明显增多，见于纤维间隔周围，终止造模后2周、即第6周时阳性染色显著强于造模4周，大量出现于汇管区，8周时较6周有所减少，表达量基本与4周时相等。阳性染色图像分析、光镜下阳染细胞计数及western blot三者的结果呈一致性。Western blot结果与免疫组织化学观察结果也具一致性。电镜下显示HOC具有形态小，核以卯圆型为主，高的核／浆比例等特点。结论 Thy1．1阳性的HOC可能与肝硬化的消减或逆转有关。     

C-myc基因和P16基因在肝细胞癌中的表达及其临床意义

目的探讨C-myc基因和P16基因表达在肝细胞癌(HCC)中的临床意义。方法采用免疫组化检测技术检测不同肝癌组织、癌旁组织及正常肝组织中的C-myc基因和P16基因表达产物。结果C-myc基因表达产物在肝癌组织中的表达明显高于癌旁组织和正常肝组织(P<0.05)。P16基因表达产物则在正常肝组织中和癌旁组织中表达明显高于肝癌组织(P<0.05)。C-myc基因的表达与HCC的发病年龄、性别、肿瘤大小、UICC分期无关(P>0.05),而与HCC的转移有关(P<0.05);P16基因的表达与HCC的发病年龄、性别、肿瘤大小无关(P>0.05),而与HCC的转移和UICC分期有关(P<0.05)。结论C-myc基因表达增强与P16基因表达障碍能促进HCC的发生发展,并可用来判断预后。     

应用抑制性消减杂交方法克隆人肝细胞癌差异表达基因

Diatchenko等[1]报道了一种新的快速克隆差异表达基因cDNA的方法,称为抑制性消减杂交(SSH),十分适用于克隆造成某种特殊细胞表型的特异表达基因,例如正常和病变之间的基因表达差异分析。人肝细胞癌(HCC)是我国高发的恶性肿瘤之一,其发生发展与多种基因的突变与表达异常有关。我们应用抑制性消减杂交方法克隆HCC与癌旁非癌组织之间差异     

GRIM-19在肝细胞癌组织中的表达及临床意义

目的:探讨GRIM-19在肝细胞癌组织中表达的临床意义.方法:用RT-PCR检测40例肝细胞癌及非癌组织中GRIM-19 mRNA表达水平,用免疫组织化学pv二步法和Western blot检测40例肝细胞癌及非癌组织中GRIM-19蛋白的表达情况,分析GRIM-19与临床病理特征的关系.结果:GRIM-19 mRNA...     

聚集素、P糖蛋白在肝细胞癌中的表达及其相关性研究

目的探讨聚集素（Clusterin）蛋白在肝细胞癌（HCC）中的表达及其与P糖蛋白（P-glycoprotein,P-gp）表达的相关性。方法采用免疫组化S-P法检测凋亡抑制基因Clusterin在HCC、肝硬化和正常肝组织中的表达及P-gp蛋白在HCC中的表达。结果Clusterin蛋白在HCC中的阳性表达率为82．93％,在肝硬化和正常肝组织中表达阴性或弱阳性;Clusterin蛋白阳性表达与患者的年龄、性别及肝硬化无关,与Edmenson组织学分级有相关性。P-gp蛋白在肝癌组织中的阳性表达率为70．73％。在HCC中,Clusterin蛋白的表达与P-gp蛋白表达呈正相关。结论Clusterin蛋白在HCC中呈高表达,与临床耐药密切相关,有望成为肝癌靶向治疗的一个新靶点。     

P53基因网络在HBV相关肝细胞癌发生机制中的作用

肿瘤抑制基因P53在乙型肝炎病毒相关肝细胞癌的发生中起着关键作用。研究乙型肝炎病毒基因组编码蛋白对P53基因网络的调控作用,有助于阐明两者在肝细胞癌发病中的作用机制。本文重点对P53基因网络的调控、P53基因突变、凋亡调节异常及乙型肝炎病毒x蛋白对P53基因网络调控的影响进行了综述。     

星形细胞上调基因1、β-连环素及周期素D1在肝细胞癌中的表达及临床意义

目的探讨肝细胞癌(HCC)组织中星形细胞上调基因(AEG)1、β-连环素(β-catenin)及周期素D1(Cyclin D1)的表达及临床意义。方法随机选取2013年7月-2014年12月贵州省人民医院经手术及病理证实为HCC的癌组织标本和对应癌旁组织各40例,另选取8例正常肝组织作为对照组。采用免疫组化SP法检测AEG-1、β-catenin及Cyclin D1蛋白在HCC组织、对应癌旁肝组织及正常肝组织中的表达情况,并分析其表达与HCC临床病理因素的相关性。计数资料组间比较采用χ2检验或Fisher确切概率法,AEG-1、β-catenin、Cyclin D1在HCC中的相关性采用Spearman等级相关分析。结果 AEG-1、β-catenin及Cyclin D1在HCC组织及癌旁肝组织中的表达均高于正常肝组织,差异均有统计学意义(χ2值分别为7.840、4.274、8.817、4.274、9.919、4.850,P值分别为0.005、0.039、0.003、0.039、0.002、0.028)。AEG-1、β-catenin、Cyclin D1的阳性表达在性别、年龄、HBs Ag、肿瘤大小方面差异无统计学意义(P值均0.05),而在病理分化程度、肝癌TNM分期及转移方面差异有统计学意义(P值均0.05)。相关性分析显示,AEG-1蛋白的表达与β-catenin、Cyclin D1蛋白的表达呈正相关(r值分别为0.420、0.741,P值均0.01)。结论 AEG-1、β-catenin及Cyclin D1在HCC的发生、发展过程中起着重要的作用;AEG-1可能通过上调Cyclin D1、β-catenin的表达和活性而促进HCC的发生和转移;联合检测三者的表达,可作为HCC基因治疗及预后评价的重要指标。     

p73基因在肝细胞癌组织中的表达与临床意义

本文通过检测35例原发性肝细胞癌与相对应的癌旁组织中p73基因的表达及突变情况,结合临床资料,探讨肝细胞癌组织中p73基因的表达与临床预后的关系。     

Prognosis of hepatocellular carcinoma expressing cytokeratin 19: Comparison with other liver cancers

AIM: To investigate whether expressing biliary phenotype predicted poor outcome after the surgical treatment in primary liver cancers.METHODS: Out of 204 patients that underwent liver resection due to hepatocellular carcinoma (HCC), liver specimens of 70 patients with HCC were evaluated for biliary components by cytokeratin (CK) 19 immunostain (CK19^- HCC and CK19⁺ HCC). CK19 positivity was defined as membranous and/or cytoplasmic expression in ≥ 5% of tumor cells with moderate or strong intensity. Patients with other primary liver cancers, such as combined HCC and cholangiocarcinoma (cHCC-CC), intrahepatic cholangiocarcinoma (ICC) who received curative liver resection, were also included in the study. Clinical characteristics of CK19^- HCC and CK19⁺ HCC patients, including survival outcome after curative liver resection, were compared with that of cHCC-CC and ICC patients.RESULTS: The overall survival (OS) rate of CK19^- HCC (n = 49) after the curative surgical treatment was 90.7%, and 80.4% at 1 and 5 years after the resection. OS rate of CK19⁺ HCC (n = 21) was 74.3%, 28.9% and OS rate of cHCC-CC (n = 22) was 66.7%, 32.2% at 1 and 5 years after the surgery. For ICC (n = 19), 1 and 5-year-OS rate was 50.2% and 14.3% after the curative resection. The OS rates of CK19⁺ HCC and cHCC-CC were significantly lower than that of CK19^- HCC, but higher than the OS rate of ICC (P = 0.000). There was no statistically significant difference in OS rate between CK19⁺ HCC and cHCC-CC. The disease free survival (DFS) rate of CK19^- HCC was 72.0% and 54.5% at 1 and 3 years after the surgical treatment. DFS rate of CK19⁺ HCC was 53.3%, 34.3% and DFS rate of cHCC-CC was 51.5%, 39.2% at 1 and 3 years after the resection. For ICC, 1 and 3-year-DFS rate was 28.0% and 14.0% after the curative resection. DFS rate of CK19^- HCC was significantly higher than that of ICC (P = 0.017), but marginally higher than DFS rate of either CK19⁺ HCC or cHCC-CC (P = 0.097, P = 0.089, respectively). Predictors of outcome after the surgery of primary liver cancer were pathology of the resected mass, existence of microvascular invasion and accompanying satellite nodule.CONCLUSION: Primary liver cancers with biliary components tended to show poorer surgical outcome. This suggested that immuno-phenotype of liver cancers was as important as their morphological classification.     

P27基因在原发性肝癌中的表达及其与预后的关系

目的　探讨 P2 7在原发性肝癌中表达 ,揭示其与原发性肝癌临床病理指标和预后的关系。方法　采用免疫组化二步法 ,检测 4 3例原发性肝癌标本、2 1例肝硬化标本和 16例正常肝脏组织中的 P2 7表达情况 ,并对2 9例肝癌患者进行了随访。结果　 P2 7在正常肝脏组织中少有表达 ,肝硬化组阳性表达有所增加 ,但两组之间差异无显著性 (P>0 .0 5 ) ;P2 7在原发性肝癌组中阳性表达率为 86 .0 4 % ,与肝硬化组和正常对照组相比差异有显著性(P<0 .0 1)。肿瘤直径 >5 cm、多个瘤灶、低分化和有血管侵犯的原发性肝癌组织中 P2 7呈低表达 ;P2 7高表达的原发性肝癌患者较低表达者生存期明显延长 (P<0 .0 1)。结论　 P2 7基因在原发性肝癌组织中的表达有组织特异性 ;P2 7与原发性肝癌的浸润和转移有密切关系 ,可以作为原发性肝癌的预后判断指标之一     

层粘连蛋白诱导体外生长的肝癌细胞表达细胞角蛋白19

目的　观察层粘连蛋白 (LN)对细胞角蛋白 (CK) 19异常表达的诱导作用 ,进一步探讨肝实质细胞额外表达CK19的分子机制。方法　对选择的 3株肝细胞癌 (HCC)细胞系的CK 19和LN的表达进行免疫酶细胞化学、免疫印迹和激光共聚焦显微镜观察 ;用含有LN的培养液进行CK19表达诱导 ,应用LN的多克隆抗体阻断这一效应。结果　 3株细胞系中有 1株产生内源性LN ,并且表达CK19,其余 2株这两种分子的表达均为阴性。当接种于含有外源性LN的培养液中生长时 ,原来CK阴性的 2株HCC细胞系均出现CK19的表达 ,其水平与LN剂量相关。LN的多克隆抗体可完全阻断外源性及内源性LN对CK19表达的诱导作用。结论　LN确实能够诱导体外生长的HCC细胞表达CK19,这种效应可被LN抗体所阻断     

P16、E-CD联合表达与乳腺癌侵袭转移关系的研究

为研究 P1 6 、E- c1adherin(E- CD)基因在乳腺癌中的联合表达及临床意义 ,应用免疫组织化学方法(SABC法 )检测了 92例乳腺癌患者癌组织 P1 6、E- CD基因蛋白的表达 ,探讨其与乳腺癌腋淋巴结和远处转移的关系。结果显示 P1 6 、E- CD基因表达与腋淋巴结转移及远隔转移有关 ;提示 P1 6 、E- CD联合基因表达在乳腺癌病程中起重要作用 ;检测其表达情况 ,对筛选乳腺癌术后高危转移患者、加强诊治有临床意义     

原发性肝细胞癌及癌旁组织中环氧合酶-2的表达及其意义

目的：研究人原发性肝细胞癌（HCC）组织和癌旁非瘤组织中的环氧合酶－2（COX－2）蛋白及基因表达情况,方法：采用免疫组织化法和原位分子杂交法,研究27对原发性肝细胞癌及癌旁非肿瘤组织,5例正常肝组织中COX－2的蛋白和基因表达,结果：高分化HCC中COX－2蛋白表达显著高于中分化和低分化HCC（P分别＜0．05）以及癌旁组织和正常组织（P分别＜0．01）,癌旁组织的COX－2表达显著高于正常组织（P＜0．05）,癌旁组织,中分化和低分化HCC之间COX－2的表达强度差异无显著性（P＞0．05）,在COX－2蛋白阳性的肝癌细胞和癌旁肝细胞胸胞胞质中可见到COX－2mRNA呈阳性表达,结论：COX－2的过度表达可能参与了高分子HCC的致癌过程。     

Expression of β-catenin in hepatocellular carcinoma

AIM: The β-catenin has been recognized as a critical member of the Wnt signaling pathway and plays an important role in the generation/differentiation of many tissues. Inappropriate activation of this pathway has been implicated in carcinogenesis. The mechanism underlying the development as well as its prognosis of hepatocellular carcinoma (HCC) has remained unclear. The purpose of this study is to analyze the expression of β-catenin in HCC in relation to histological grades and viral hepatitis backgrounds.METHODS: Thirty-two sections were selected at random from autopsy and surgical cases of HCC. Immuohistologically,the location and positivity of β-catenin expression in HCC was examined.RESULTS: Normal hepatocytes did not express β-catenin.In 78% of HCC β-catenin was expressed at the membrane of the cells, with or without cytoplasmic and/or nuclear expression. The tumor cells with well- and moderatelydifferentiated grades expressed frequently at the membrane and cytoplasm compared with poorly-differentiated type.Nuclear expression of β-catenin was prone to occur in the tumor cells of poorly-differentiated grade. There were 15% of hepatitis C virus (HCV) backgrounds with nuclear expression. In contrast, there was 38% with nuclear expression in hepatitis B virus (HBV) backgrounds. In nonBnonC hepatitis, no case expressed nuclear β-catenin.CONCLUSION: The β-catenin expression in HCC cells was heterogenous among types of hepatitis viral infection.Wnt signaling pathway might be deeply involved in less-differentiated HCC and HBV background.     

细胞角质蛋白19与肝癌临床病理关系的探讨

目的 分析原发性肝细胞癌(HCC)患者血清中细胞角质蛋白(CK)19水平与临床病理指标的关系。方法 用放射性核素标记的免疫吸附试验(ELISA)检测101例正常人血清中CK19浓度,确定参考范围;然后测定108例HCC患者术前血清CK19浓度,分析CK19升高者的临床病理特征。结果 正常人血清CK19浓度单侧98％可信区间上限是2.3μg/L。108例HCC患者中,24例(22.2％)CK19水平升高,浓度范围是2.4～4 5.5μg/L,其中12例(11.1％)只有CK19升高而AFP正常。低分化癌的比例在CK19升高者中(37.5％,9/24)高于CK19正常者(20.2％,17/84,x~2=7.362,P<0.05)。门静脉癌栓的比例在CK19升高组(25.0％,6/24)高于CK19正常组(6.0％,5/84,x~2=7.403,P<0.01);TNM Ⅲ/Ⅳ期肿瘤的比例在CK19升高组(54,2％,13/24)亦高于CK19正常组(21.4％,18/84,x~2=13.300,P<0.005)。结论 部分HCC患者血清中CK19升高,可能与肿瘤门静脉癌栓、分化程度更低、病期更晚有关。     

Expression of metastasis suppressor gene nm23 in human hepatocellular carcinoma

AIM: To investigate the relationship between the expression of nm23-Hi mRNA and the metastatic potential of hepatocellular carcinoma (HCC). METHODS: The expression of nm23-H1 mRNA was detected in 24 cases of HCC by in situ hybridization using digoxigenin-labeled nm23-H1 antisense cRNA probe. Twenty-four HCC specimens were divided into two groups according to the following criteria: (1) metastasis in portal lymph nodes; (2) the number of tumors in the liver; (3) cancerous emboli in the portal vein; and (4) the existence of satellite lesions. We named those meeting criteria (1) or (2) and (3), or (3) and (4) high metastatic potential (n = 6); and the others formed the low metastatic potential group (n = 18). RESULTS: Positive results of in situ hybridization showed granules or masses in the cytoplasm. In the low metastatic potential group strong staining was obtained in ten specimens, while in the high metastatic potential group there was none. Three negative results were found in the high metastatic potential group, and one in the low metastatic potential group (P < 0.05). The expression of nm23-H1 mRNA was not correlated with some clinical factors, such as tumor size or the background liver disease. CONCLUSION: The expression of nm23-H1 mRNA is inversely correlated with HCC metastatic potential, and can be considered as an index which indicates the metastatic potential of HCC.     

原发性肝癌组织中DLK1表达及意义

目的探讨DLK1在原发性肝细胞性肝癌（HCC）发生、发展中的作用。方法分别应用免疫组化sP法和原位杂交的方法检测150例原发性肝细胞癌、50例肝硬化、50例肝炎组织中和10例意外死亡（生前体健）者肝组织中DLK1表达。结果DLK1蛋白及DLK1mRNA在HCC中的表达明显高于肝硬化、肝炎和正常肝组织表达（P均〈0。05）。DLK1蛋白表达与肿瘤直径及临床分期有明显相关性（P均〈0．05）。结论DLK1高表达可促进HCC的发生、发展。