肝癌组织及血清中细胞间粘附分子-1表达的研究

目的研究细胞间粘附分子－１（ｉｎｔｅｒｃｅｌｕｌａｒａｄｈｅｓｉｏｎｍｏｌｅｃｕｌｅ－１，ＩＣＡＭ－１）作为判断肝癌发展程度及转移状态指标的可能性。方法以免疫组化的方法检测ＩＣＡＭ－１在肝癌组织和正常肝组织中的表达，观察了ＩＣＡＭ－１表达在细胞的定位。以点免疫印迹法测定不同患者血清和不同肝组织中ＩＣＡＭ－１的表达。分析ＩＣＡＭ－１表达与肿瘤生长转移状态、肿瘤特性的关系。结果肝癌细胞ＩＣＡＭ－１表达阳性（阳性率为８００％），主要分布在细胞膜，正常肝细胞则为阴性。肝癌患者血清可溶性细胞间粘附分子－１（ｓＩＣＡＭ－１）水平高于正常人（Ｐ＜００１）及肝良性肿瘤患者（Ｐ＜００５），肝癌伴转移者高于无转移者（Ｐ＜００５）。肝癌组织中ＩＣＡＭ－１含量明显高于癌旁组织（Ｐ＜００１）及正常肝组织（Ｐ＜００１），而与肿瘤大小及有无包膜无关（Ｐ＞００５）；转移组肝癌中ＩＣＡＭ－１的表达明显高于非转移组（Ｐ＜００５），两组癌旁组织中ＩＣＡＭ－１表达无差别（Ｐ＞００５）。结论血清及组织中ＩＣＡＭ－１的水平在一定程度上可以反映肝癌发展程度及转移状态，有可能作为预测肝癌转移复发的指标。     

细胞间粘附分子-1在乙型肝炎后肝细胞癌表达的免疫组化研究

为探讨细胞间粘附分子－１（ＩＣＡＭ－１）在原发性肝细胞癌的表达状况及其在肝癌发生中的作用，利用免疫组织化学技术链菌素亲生物蛋白（ＬＳＡＢ）法检测３０例乙型肝炎后肝细胞癌及癌旁肝组织ＩＣＡＭ－１的表达情况，结果 肝癌组织中，ＩＣＡＭ－１存在一定程度的表达，其中癌组织阳性率为４３．３％，多为弱表达，癌旁肝组织明显高于癌组织，阳性率为７８．６％，多呈弥漫强阳性表达，二者差异有显著性（Ｐ〈０．１０）。结论     

细胞间粘附分子－1在乙型肝炎中的表达及意义

为观察细胞间粘附分子－１（ＩＣＡＭ－１）在乙型肝炎肝组织内的表达状况，探讨ＩＣＡＭ－１在乙型肝炎肝细胞免疫损伤中的作用。采用链菌素亲生物蛋白法（ＬＳＡＢ），以ＩＣＡＭ－１的单克隆抗体，检测１０４例急、慢性乙型肝炎，９例无症状携带者，１０例正常肝组织标本内ＩＣＡＭ－１抗原表达情况。结果发现：ＩＣＡＭ－１在正常肝细胞无表达，肝窦内皮细胞弱表达；ＨＢＶ感染后肝细胞呈不同程度的表达，肝窦内皮表达增强；中、重度慢性肝炎，活动性肝硬化肝细胞ＩＣＡＭ－１表达较急性肝炎，轻度慢性肝炎显著增强（Ｐ＜０．０１）；急性肝炎较轻度慢性肝炎显著增强（Ｐ＜０．０１）；轻度慢性肝炎较无症状携带者，正常肝组织显著增强（Ｐ＜０．０１）。表明ＩＣＡＭ－１在乙型肝炎肝细胞内表达与肝细胞损伤有关，对ＨＢＶ的清除可能起着重要作用，ＩＣＡＭ－１在肝窦内皮的表达有助于淋巴细胞向肝组织内浸润。     

细胞间粘附分子-1在肝硬化及肝癌组织中的表达

对３０例原发性肝癌及２３例肝炎肝硬化组织细胞间粘附分子一１（ＩＣＡＭ－１）的表达进行原位检测,探讨了ＩＣＡＭ－１在肝癌发生免疫逃逸机制中的意义。１．标本收集：肝炎肝硬化标本２３例,为乙肝患者肝穿组织。原发性肝癌标本３０例,为肝穿组织及手术病理存档标本,病理诊断为肝细胞癌,本组病例ＨＢＳＡｇ均阳性,癌旁肝组织呈慢性炎症并伴有不同程度肝硬化改变。标本均进行连续切片分别用于ＨＥ染色及ＩＣＡＭ－ｌ检测。２．主要试剂及方法：ＩＣＡＭ一１单克隆抗体由美国波斯顿血液研究中心馈赠,ＬＳＡＢ试剂盒为ＤａＫｏ公司产…     

原发性肝细胞癌患者转化生长因子β1的基因表达及临床意义

目的研究转化生长因子β１（ＴＧＦβ１）ｍＲＮＡ在原发性肝细胞癌患者中的表达及其临床意义。方法用ＲＴ－ＰＣＲ加ＤｏｔＢｌｏｔ法检测原发性肝细胞癌患者肝组织和外周血单个核细胞（ＰＢＭＣ）中ＴＧＦ－β１ｍＲＮＡ水平，并以正常肝组织和正常人的ＰＢＭＣ为对照。结果ＴＧＦ－β１ｍＲＮＡ水平在原发性肝细胞癌患者组肝组织（２．２２±０．８４，ｎ＝１６）和ＰＢＭＣ中（１．８３±１．２，ｎ＝２５）比较，差异无显著性（Ｐ＞０．０５），但两者均高于正常肝组织（０．９４±０．７６，ｎ＝５）和正常人的ＰＢＭＣ（０．６２±０．４０，ｎ＝１６）水平。结论ＴＧＦ－β１ｍＲＮＡ水平与原发性肝细胞癌有关，ＰＢＭＣ中ＴＧＦ－β１ｍＲＮＡ检测可望作为一项代替肝组织活检的指标，其表达水平与肝癌有关。     

脑卒中患者转化生长因子与细胞黏附因子检测及分析

目的 探讨脑卒中患者外周血转化生长因子（ＴＧＦ－β１）和细胞黏附因子（ｓＩＣＡＭ－１）水平的变化并对其进行分析。方法 对脑卒中患者（脑出血３０例，脑血栓３０例）外周血ＴＧＦ－β１和ｓＩＣＡＭ－１水平进行检测。结果 脑出血患者外周血ＴＧＦ－β１的水平明显低于对照组（Ｐ〈０．０１），而ｓＩＣＡＭ－１水平明显高于对照组（Ｐ〈０．０１）；脑血栓患者外周血ＴＧＦ－β１水平与对照组差异无显著性，而ｓＩＣＡＭ－     

细胞间粘附分子1mRNA在哮喘豚鼠支气管组织中的原位表达

为了解哮喘豚鼠支气管组织中的细胞间粘附分子１（ＩＣＡＭ－１）ｍＲＮＡ表达情况，将实验豚鼠随机分为正常对照组、氟美松组和哮喘组，采用原位杂交方法显示ＩＣＡＭ－１ｍＲＮＡ的表达变化，用ＨＥ染色方法来测定炎细胞在支气管粘膜中的浸润程度。原位杂交结果显示，ＩＣＡＭ－１ｍＲＮＡ主要在支气管粘膜下的小血管、毛细血管内皮细胞以及支气管上皮细胞的胞浆内表达。定量分析结果表明，哮喘组豚鼠支气管组织表达ＩＣＡＭ－１ｍＲＮＡ较正常对照组及氟美松组显著增加（Ｐ＜０．０２，Ｐ＜０．０５），哮喘豚鼠支气管组织ＩＣＡＭ－１ｍＲＮＡ的表达量与炎细胞总数之间显著相关（Ｐ＜０．０５）。提示支气管粘膜组织ＩＣＡＭ－１ｍＲＮＡ表达与哮喘支气管粘膜嗜酸细胞为主的炎细胞浸润有关，糖皮质激素能够抑制细胞粘附分子的表达。     

Ⅳ型胶原酶的表达与肝癌侵袭转移的关系

目的研究Ｍｒ７２０００Ⅳ型胶原酶（ＭＭＰ＿２）在肝癌细胞中的表达与肝癌侵袭转移的关系．方法以原位分子杂交及免疫组化方法检测３４例原发性肝癌及其癌旁组织中ＭＭＰ＿２ｍＲＮＡ及其蛋白的表达．患者３０例伴肝硬变，４例无肝硬变，其中２４例肝癌突破包膜向周围肝组织侵袭性生长、门静脉癌栓形成或形成转移灶，１０例肝癌包膜完整．结果呈侵袭性生长的２４例肝癌中１６例癌细胞ＭＭＰ＿２ｍＲＮＡ表达明显增高，而包膜完整的１０例肝癌中仅有２例ＭＭＰ＿２ｍＲＮＡ表达阳性．侵袭性生长的肝癌２４例中１８例ＭＭＰ＿２呈不同程度的阳性表达，而包膜完整的肝癌１０例中仅３例ＭＭＰ＿２表达阳性，向外突出生长的肝癌细胞ＭＭＰ＿２表达阳性，转移组肝癌（发生门静脉癌栓或转移）ＭＭＰ＿２阳性率（８３３％，１５／１８）明显高于未转移组肝癌ＭＭＰ＿２阳性率（３７５％，６／１６）（Ｐ＜００１）．结论由肝癌细胞产生的ＭＭＰ＿２与肝癌的侵袭转移密切相关．     

皮肤白细胞碎裂型血管炎患者皮损组织中粘附分子的表达及与病理变化的关系

应用免疫组化技术检测１８例白型血管炎（ＬＣＶ）患者皮损组织中粘附分子的表达,观察其与皮损组织病理变化的关系,并与１５例对照者进行比较。结果显示：ＬＣＶ患者血管内皮细胞的细胞间粘分子－１（ＩＣＡＭ－１）表达明显上调,血管内皮细胞粘附分子－１（ＶＣＡＭ－１）表达仅见于后期皮损,内皮白细胞粘附分子－１（ＥＬＡＭ－１）与皮损持续时间呈负相关（ｒ＝－０．９６２３,Ｐ〈０．０５）、与中性粒细胞浸润吴正相关（ｒ     

mdr-1、CD44v、和nm23-H1基因产物与肝癌侵袭及转移的关系

为探讨ｍｄｒ－１,ＣＤ４４ｖ和ｎｍ２３－Ｈ１基因产物蛋白与肝癌侵袭,转移预后的关系。用免疫组织化学技术检测４６例肝癌患者ＣＤ４４ｖ和ｎｍ２３－Ｈ１和ｍｄｒ－１基因蛋白的表达。结果 显示肝癌组织ｍｄｒ－１,ＣＤ４４ｖ和ｎｍ２３－Ｈ１（６０．０８％）,明显高于正常组织和癌旁组织（Ｐ〈０．０５）;肝癌转移组的ｍｄｒ－１,ＣＤ４４ｖ阳性表达率高于ｎｍ２３－Ｈ１阳性表达率,差异显著（Ｐ〈０．０５）;肝癌非转     

Expression of intercellular adhesive molecule1 in liver cancer tissues and liver cancer metastasis

AIM: To study the relationship between intercellular adhesive molecule-1 (ICAM-1) and liver cancer metastasis and to search for factors to predict metastasis of liver cancer.METHODS: ICAM-1 expression in fresh tissues of normal liver and hepatocellular cancer (HCC) was examined by immunoperoxidase staining. The expression of ICAM-1 in human hepatoma, tumor surrounding tissues and normal livers were semiquantitatively analyzed by Dot immuno blot. Tissue ICAM-1 expression at mRNA level was detected by Northern blot.RESULTS: All 6 cases of normal liver samples were negative in anti-ICAM-1 immunohistochemical staining, 80.0% (36/45) of HCC presented various ICAM-1 expression. The number of positive cells was a little higher in large tumors, tumors with intact capsule and metastasis, but there was no significant difference. Two cases with cancer embolus also had high ICAM-1 expression. ICAM-1 concentration in HCC (13.43 ± 0.09) was higher than that in tumor surrounding tissues (5.89 ± 0.17, P < 0.01) and normal livers (4.27 ± 0.21, P < 0.01). It was also higher in metastasis group (20.24 ± 0.30) than in nonmetastasis group (10.23 ± 0.12, P < 0.05). Northern blot analysis revealed that ICAM-1 expression at mRNA level was also higher in HCC and cancer embolus than that in tumor surrounding tissues and normal livers.CONCLUSION: Tissue ICAM-1 could indicate the growth and metastasis of HCC, and may be an index that can predict liver cancer metastasis.     

Invasion and metastasis of liver cancer: expression of intercellular adhesion molecule 1

To study the relationship between intercellular adhesion molecule 1 (ICAM-1) and liver cancer metastasis and to find predicting factors that could indicate the growth and metastasis of liver cancer. Methods: ICAM-1 expression in fresh tissue of normal liver and hepatocellular cancer (HCC) was examined by immunoperoxidase staining. Serum soluble intercellular adhesion molecule 1 (sICAM-1) from patients with a benign HCC tumor, and the expression of ICAM-1 in the orthotopically transplanted LCI-D20 tumor of a nude mouse liver cancer metastasis model, and in human hepatoma, the tumor surrounding tissue and normal liver, was analyzed semiquantitatively by the immuno-dot blot method. Tissue ICAM-1 expression (mRNA level) was detected by Northern blotting. Results: ICAM-1 expression in LD1-20 D metastatic liver cancer had a positive correlation with tumor size and the time after implantation. It increased suddenly as metastasis occurred being 3.03 ± 0.51 before metastasis and 8.24 ± 0.95 after metastasis, P < 0.01, then remained high, appending on the number of sites involved (monosite metastasis 5.48 ± 0.49, multisite metastasis 10.05 ± 1.17, P < 0.05). All six cases of normal liver samples were negative in anti-ICAM-1 immunohistochemical staining, 80.0% (36/45) of the HCC showed some ICAM-1 expression. The rate of positive cells was a little higher in large tumors, tumors with an intact capsule and tumors with metastasis, but there was no significant difference. It was noticed that two cancer emboli also had high ICAM-1 expression. The ICAM-1 concentration in HCC (13.43 ± 0.09) was higher than that in tumor surrounding the liver (5.89 ± 0.17, P < 0.01) and that in normal liver (4.27 ± 0.21, P < 0.01). sICAM-1, like tissue ICAM-1, was higher in HCC patients than in patients (with benign liver tumor and normal controls. Both tissue ICAM-1 and sICAM-1 were higher in the metastasis group than in the group without metastasis (tissue ICAM-1 20.24 ± 0.30 vs 10.23 ± 0.12 P < 0.05; sICAM-1 12.18 ± 0.25 vs 9.77 ± 0.54 P < 0.05). Northern blot analysis revealed that ICAM-1 expression, as indicated by mRNA level, was also higher in HCC and in cancer emboli than in tumor surrounding liver and normal liver. Conclusions: Tissue ICAM-1 and serum sICAM-1 could indicate the stage of HCC, and the potential of hepatoma cells for invasion and metastasis. They may play an important role in the metastasis cascade. Received: 20 January 1998 / Accepted: 25 June 1998     

Phase tissue intercellular adhesion molecule-1 expression in nude mice human liver cancer metastasis model

Ｐｈａｓｅｔｉｓｓｕｅｉｎｔｅｒｃｅｌｕｌａｒａｄｈｅｓｉｏｎｍｏｌｅｃｕｌｅ１ｅｘｐｒｅｓｓｉｏｎｉｎｎｕｄｅｍｉｃｅｈｕｍａｎｌｉｖｅｒｃａｎｃｅｒｍｅｔａｓｔａｓｉｓｍｏｄｅｌＳＵＮＪｉｎｇＪｉｎｇ,ＺＨＯＵＸｉｎＤａ,ＬＩＵＹｉｎＫ...     

Phase tissue intercellular adhesion molecule-1 expression in nude mice human liver cancer metastasis model

AIM To study the phase cancer tissue intercellular adhesion molecule-1 (ICAM-1) expression of human cancer metastasis model in nude mice, and to analyze the relationship between ICAM-1 expression and the metastasis and recurrence of hepatocellular cancinoma (HCC).METHODS HCC tissues from liver cancer metastasis model in nude mice (LCI-D20) was orthotopically implanted, and ICAM-1 expression in HCC tissues at different growing time were detected by immunodot blot. Tumor size, intrahepatic and extrahepatic metastasis foci were observed by naked eyes and under light microscope.RESULTS ICAM-1 was positively correlated to the tumor growing time (r=0.88, P＜0.01) and tumor size r=0.5, P＜0.05). It was higher in metastatic HCC than in nonmetastatic HCC (8.24±0.95 vs 3.03±0.51, P＜0.01). ICAM-1 content in cancer tissues increased suddenly after metastasis occurred and then maintained in a high level. ICAM-1 was also higher in multimetastasis group than in monometastasis group (10.05±1.17 vs 5.48±0.49, P＜0.05).CONCLUSION Tissue ICAM-1 could predict not only the metastasis of human liver cancer metastasis model in nude mice early and sensitively, but also the metastasis degree. So tissue ICAM-1 may be a potential index indicating the status of metastasis of HCC patients.     

Tissue microarray for high-throughput analysis of gene expression profiles in hepatocellular carcinoma

AIM: To study the expression profiles of HBsAg, HBcAg, p21WAF1/CIP1 (p21), Rb genes in hepatocellular carcinoma (HCC) and to investigate their roles in the hepatocar-cinogenesis. METHODS: HCC tissue microarray containing 120-min tissues of 40 HCC cases was constructed. HBsAg, HBcAg, p21 and Rb proteins were immunohistochemically stained by streptavidin-peroxidase conjugated method (S-P). The expression loss of these genes in cancerous, para-cancerous tissues and adjacent normal liver tissues of 40 HCCs were comparatively examined. RESULTS: The positive rate of HBsAg expression in cancerous tissues of 40 HCCs was 7.5%, which was lower than that in para-cancerous and adjacent normal liver tissues (X2=12.774, P<0.01; X2=18.442, P<0.01). The positive rate of HBcAg expression in cancerous tissues of 40 HCCs was 20.0%, which was also lower than that in para-cancerous and adjacent normal liver tissues (X2=9.482, P<0.01; X2=14.645, P<0.01). p21 protein deletion rate in cancerous tissues of 40 HCCs was 27.5%, which was higher than that in para-cancerous and adjacent normal liver tissues (X2=7.439, P<0.01; X2=11.174, P<0.01). p21 protein deletion correlated remarkably with the pathological grade of HCC (X2=0.072, P<0.05). Rb protein deletion rate in cancerous tissues of 40 HCCs was 42.5%, which was also higher than that in para-cancerous and adjacent normal liver tissues (X2=10.551, P<0.01; X2=18.353, P<0.01). Rb protein deletion rate did not correlate remarkably with tumor size or pathological grade of HCC (X2=0.014, P>0.05; X2=0.017, P>0.05). CONCLUSION: Expression deletion of HBsAg, HBcAg, p21 and Rb proteins in HCCs may play important roles in the carcinogenesis of HCC. Tissue microarray is an effective high-throughput technique platform for cancer research.     

Overexpression of p28／gankyrin in human hepatocellular carcinoma and its clinical significance

AIM：To investigate the expression of p28/gankyringene and its role in the carcinogenetic process of human hepatocellular carcinoma(HCC).METHODS:64specimens of HCC and para-carcinoma tissues,22specimens of non-tumor liver tissues(7normal,15cirrhosis),10 specimens of normal human tissues and 5hepatoma cell lines were studied for the expression of p28/gankyrinby Northern blot.The expressionof p28/gankyrin protein was detected immunohistochemically by using the specific polyclonal antibody.RESULTS:Northern blot analysis indicated that the expression of p28/gankyrinmRNA was intensively distributed in brain and heart,weakly in lung,spleen and muscle,undetectable in digestive system including liver,pancreas,stomach,small and large intestines.p28/gankyrinmRNAwas absent in normal liver,weakly detected in liver cirrhosis and in 18of 64para-carcinoma liver tissues.In contrast,the expression of p28/gankyrinmRNA was intensitely detected in all5hepatoma cell lines tested,markedly increased in 57of 64and moderately increased in 5of64HCCsamples.In comparison with liver cirrhosis and para-carcinoma liver tissues.the average expression of p28/gankyrinmRNAin HCCwas increased3.6-(2.901&#177;0.507vs0.805&#177;0.252,P&lt;0.05)and5.2-fold(2.901&#177;0.507vs0.557&#177;0.203,p&lt;0.01),respectively.In addition,p28/gankyrinmRNA expression level was higher in HCCwith portal vein tumor thrombus and microscopic hepatic vein involvement(P=0.021and P=0.047,respectively).Theoverexpression of p28/gankyrin protein in HCCwas targeted in hepatic tumor cells,not in bile duct cells and other interstitial cells.Plays an important role and contributes to the metastasis potential in the process of carcinogenesis.p28/gankyrinmay become a specific biological tissue marker for the pathological diagnosis of HCC.     

巨噬细胞移动抑制因子和细胞周期蛋白D1的表达与肝细胞癌生长和转移的关系

目的研究巨噬细胞移动抑制因子（MIF）、细胞周期蛋白D1（Cyclin D1）、细胞周期蛋白依赖激酶4（CDK4）、磷酸化视网膜母细胞瘤易感基因产物蛋白（phospho—Rb）在HeC组织中的表达及其与癌细胞生长和转移的关系。方法应用组织芯片技术和免疫组织化学方法检测93份HCC组织中MIF、Cyclin D1、CDK4和phospho—Rb的表达，分析它们的表达与HCC临床病理特征的关系。结果93份HCC组织中MIF、Cyclin D1、CDK4和phospho—Rb的表达率分别为71％、41％、82％和14％，MIF和Cyclin D1阳性表达率与正常肝脏组织表达率之间的差异有统计学意义（P〈0．01），CDK4和phospho—Rb阳性表达率与正常肝脏组织表达率之间的差异无统计学意义（P〉0．05）。在≥3．5cm的肿瘤中MIF表达率为79％，明显高于在〈3．5cm肿瘤中的表达率48％（P〈0．01）；有转移的肝癌组织中Cyclin D1阳性率为62％，明显高于无转移组织中的阳性率35％，差异有统计学意义（P〈0．05）。MIF表达强弱与Cyclin D1的表达呈正相关关系（P〈0，01）。CDK4和phospho—Rb的表达与肿瘤大小及是否转移的关系无统计学意义。结论MIF和Cyclin D1在HCC发展进程中可能促进肿瘤的生长和转移。