人肝细胞癌中整合素α5,β1亚基和纤维连接蛋白mRNAs的表达

目的研究肝细胞癌（ＨＣＣ）中整合素α５、β１亚基和纤维连接蛋白（ＦＮ）ｍＲＮＡｓ表达的生物学意义。方法应用Ｎｏｒｔｈｅｒｎ印迹杂交和核酸原位杂交技术，对１５例ＨＣＣ癌组织、５例癌旁肝硬化和３例正常肝组织中整合素α５、β１亚基和ＦＮｍＲＮＡｓ表达作杂交分析。结果高分化ＨＣＣ癌组织中三种ｍＲＮＡｓ表达水平与癌旁及正常肝组织相近，而低、中分化ＨＣＣ癌组织内明显减少，甚至消失（分别Ｐ＜００１）；整合素α５亚基和ＦＮｍＲＮＡｓ主要见于癌细胞胞浆内；ＨＣＣ伴肝内浸润和／或转移组癌组织内三种ｍＲＮＡｓ水平较无浸润转移组显著下降（Ｐ＜００５）；５例伴肝内转移的ＨＣＣ癌组织中ＦＮｍＲＮＡ呈异质性表达。结论ＨＣＣ中整合素α５、β１和ＦＮ蛋白的表达变化系癌细胞基因转录水平下调的结果，可能与ＨＣＣ细胞分化、浸润和转移相关；异质性ＦＮｍＲＮＡ及其编码的ＦＮ蛋白可能在ＨＣＣ转移中有意义。     

大鼠肝纤维化Ito细胞纤维连接蛋白受体α5β1的表达

研究纤维连接蛋白受体α５β１在大鼠肝纤维化中的作用。     

肝细胞癌nm23—H1蛋白表达的研究

应用免疫组化检测８８例肝细胞癌中ｎｍ２３－Ｈ１蛋白的表达，癌旁肝组织强阳性表达，５１例肝癌组织阳性表达（５８％），阳性产物主要定位于肿瘤细胞胞浆。ｎｍ２３－Ｈ１蛋白表达与ＨＣＣ肿瘤体积，组织分型及Ｅ ｄｍｏｎｄｓｏｎ分级无关，而与肝内或肝外转移显著负相关，结果表明ｎｍ２３－Ｈ１在抑制ＨＣＣ肝内或肝外转移起着重要作用，有可能成为评价ＨＣＣ病人预凰的一项新指标。     

肝细胞癌中DEK蛋白的表达及其意义

目的 探讨DEK蛋白在肝细胞癌(hepatocellular carcinoma,HCC)中的表达及意义.方法 应用组织芯片技术进行免疫组化染色检测92例HCC和58例癌旁肝组织标本中DEK的表达,结合临床相关因素进行统计分析.结果 组织芯片检测结果中实际有效146个,其中HCC组织88例,癌旁肝组织58例.HCC、癌旁肝组织中DEK阳性表达率分别为39.8%、5.2%,HCC组织中DEK阳性表达率明显高于癌旁肝组织(P=0.000);HCC组织中DEK阳性表达与组织分化程度、是否转移密切相关(P<0.05).结论 HCC组织中DEK蛋白的表达对其诊断及预后有一定参考价值.     

脑胶质瘤中纤维连接蛋白及其整合素β1受体的表达

目的探讨脑胶质瘤中纤维连接蛋白（FN）及其整合素β1受体蛋白（β1 integrin）表达的临床病理联系。方法用免疫组化S-P法检测56例胶质瘤组织中FN和整合素β1的表达情况,并进行相关临床病理分析。结果（1）胶质瘤组织问质血管壁FN染色随级别增高而增厚（P〈0.01）,且血管厚度在复发病例组明显高于无复发组（P〈0．05）。恶性程度高的Ⅲ级与Ⅳ级胶质瘤组织中还可见基质弥散阳性染色。（2）FN和整合素β1在胶质瘤中阳性表达率分别为30．36％（17／56）和39．29％（22／56）,并均随肿瘤级别增加而增高（P〈0.05,P〈0．01）,两者表达存在相关性（rs=0．312,P〈0．05）。复发病例FN和整合素β1阳性表达率（68．42％,57．89％）均高于无复发组（10．81％,29．73％）（P〈0．01,P〈0．05）。结论FN及其整合素β1受体在胶质瘤组织中的高表达促进了胶质瘤的恶性进展,两者可能成为有价值的判断胶质瘤恶性程度及预后的指标。     

肝细胞癌中EMS1蛋白的表达

目的 探讨EMS1蛋白表达与肝癌发生、发展及临床病理特征的关系.方法 采用免疫组化SP法,对78例肝癌组织、其中68例附癌旁肝组织和8例正常肝组织中EMS1蛋白表达进行检测.结果 EMS1蛋白表达定位于细胞的胞质,着色呈黄色至棕黄色.肝癌中阳性表达率89.69%,高于癌旁及正常肝组织30.26%(P＜0.001);无肝硬化肝癌阳性表达率73.91%,低于伴有肝硬化95.56% (P＜0.05);伴静脉内癌栓组94.12%与无静脉内癌栓组70.59%差异有显著性 (P＜0.05);随着组织分级的增高(分化程度的降低),阳性表达率明显增高,其差异有显著性 (P＜0.05);而与性别、年龄、肿瘤大小无关.结论 EMS1蛋白高表达与肝癌发生、肝硬化、静脉内癌栓及组织学分级相关.     

细胞周期蛋白G1在喉鳞状细胞癌中的表达及其临床病理学意义

目的 探讨喉鳞状细胞癌组织中细胞周期蛋白(cyclin)G1表达与临床病理参数及p53、癌组织微血管密度(MVD)的关系,并分析其作为预测喉鳞状细胞癌预后指标的可能性.方法 采用免疫组织化学(EnVision法)检测81例喉鳞状细胞癌组织中cyclin G1、p53的表达情况,用CD34标记新生血管内皮细胞,在显微镜下观察MVD.同时测它们在20例癌旁正常喉黏膜中的表达.结果 与正常喉黏膜组织相比,肿瘤组织中cyclin G1、p53表达及MVD均明显增加.喉鳞状细胞癌组织中cyclin G1、p53的阳性表达率分别为61.7%(50/81)和65.4%(53/81),在p53蛋白表达阳性和阴性的喉鳞状细胞癌组织标本中,cyclin G1蛋白阳性率分别为71.7%(38/53)和42.9%(12/28),cyclin G1表达与p53有相关性(Kappa值为0.281,P=0.011).cyclin G1蛋白阳性组平均MVD为(51.23±16.46),阴性者(30.74±12.29),其差异有统计学意义(P=0.005).cyclin G1高表达与肿瘤组织分化程度、颈淋巴结转移以及5年生存率均有统计学意义(P值分别为0.002、0.013和0.032).结论 cyclin G1蛋白在喉鳞状细胞癌中存在高表达,并与p53、MVD关系密切,cyclin G1异常表达可能在喉鳞状细胞癌发生、发展及转移过程中发挥重要作用,并可能作为判断患者预后的指标之一.     

肝癌凋亡细胞相关蛋白的表达分析

目的；分析肝癌凋亡细胞与肝癌细胞所表达的蛋白质在分子量分布上的差异。方法：ＳＤＳ＿ＰＡＧＥ电泳后薄层扫描。结果：肝癌凋亡细胞所表达蛋白质的分子量普遍较大。在分子量大于６．７×１０＾４ｕ时，凋亡的肝癌细胞有７个较为明显的蛋白扫描峰，与未凋亡的肝癌细胞具有明显的区别。部分大分子量的凋亡相关蛋白在肝癌凋亡细胞表达量很高，其蛋白质扫描峰的面积可占总面积的８％。结论：肝癌凋亡细胞有较大分子量的蛋白质表达，可     

乳腺浸润性癌PS2蛋白的表达及其与预后的关系

目的探讨ＰＳ２蛋白在乳腺浸润性癌中的表达及其与预后的关系。方法应用ＬＳＡＢ免疫组化法检测ＰＳ２蛋白在８６例乳腺浸润性癌中的表达。结果ＰＳ２蛋白的表达率为６６．２７％（５７／８６）。在下列一些情况中，ＰＳ２蛋白的表达率有区别：（１）８６例中，５年以上组８０．５５％（２９／３６），５年以下组５６．００％（２８／５０），差异有显著性意义（Ｐ＜０．０２５）；（２）未停经组６２例，５年以上组８６．２０（２５／２９），５年以下组５４．５４％（１８／３３），差异有显著性意义（Ｐ＜０．００５），已停经组２４例，５年以上组４／７例，５年以下组５８．８０％（１０／１７），差异无显著性意义；（３）腋窝淋巴结转移６２例，５年以上组８２．３５％（１４／１７），５年以下组５５．５５％（２５／４５），差异有显著性意义（Ｐ＜０．０５）；腋窝淋巴结阴性２４例，５年以上组７８．９４％（１５／１９），５年以内组３／５例，差异无显著性意义。结论本研究表明，ＰＳ２蛋白的阳性表达与５年生存期正相关，ＰＳ２阳性的表达可以作为乳腺癌的一项预后指标；ＰＳ２蛋白表达对未停经患者的内分泌治疗具有一定的指导意义；腋窝淋巴结阳性患者ＰＳ２蛋白表达与较好的预后相关。     

Engineering cell adhesive surfaces that direct integrin alpha5beta1 binding using a recombinant fragment of fibronectin

Integrin receptors mediate cell adhesion to extracellular matrices and trigger signals that direct cell function. While many integrins bind to the arginine-glycine-aspartic acid (RGD) motif present in numerous extracellular proteins, integrin alpha(5)beta(1) requires both the PHSRN synergy site in the 9th and the RGD site in the 10th type III repeat of fibronectin (FN). Binding of alpha(5)beta(1) to FN is critical to many cellular processes, including osteoblast and myoblast differentiation. This work focused on engineering integrin-specific bioadhesive surfaces by immobilizing a recombinant FN fragment (FNIII(7-10)) encompassing the alpha(5)beta(1) binding domains of FN. Model hybrid surfaces were engineered by immobilizing FNIII(7-10) onto passively adsorbed, non-adhesive albumin. Homo- and hetero-bifunctional crosslinkers of varying spacer-arm length targeting either the cysteine or lysine groups on FNIII(7-10) were investigated in ELISA and cell adhesion assays to optimize immobilization densities and activity. FN-mimetic surfaces presenting controlled densities of FNIII(7-10) were generated by varying the concentration of FNIII(7-10) in the coupling solution at a constant crosslinker concentration. Cells adhered to these functionalized surfaces via integrin alpha(5)beta(1) and blocking with integrin-specific antibodies completely eliminated adhesion. In addition, adherent cells spread and assembled focal adhesions containing alpha(5)beta(1), vinculin, and talin. This biomolecular engineering strategy represents a robust approach to increase biofunctional activity and integrin specificity of biomimetic materials.     

CAS基因在原发性肝癌组织中的表达及其与HBV感染的关系

目的 探讨细胞凋亡易感基因(CAS)可否作为肝癌的病理诊断标志物,以及肝细胞癌中CAS蛋白的表达与HBV感染之间的关系.方法 应用免疫组化法检测肝癌、癌旁组织及未发生肿瘤的肝硬化、肝炎组织中CAS蛋白的表达情况,同时应用免疫组化法、核酸原位杂交法检测HBV感染的肝细胞癌组织、癌旁组织中HBsAg、HBcAg以及HBV DNA的表达情况,分析肝细胞癌中CAS蛋白的表达与HBV感染之间的关系.结果 CAS蛋白在肝癌组织中的表达较癌旁组织明显升高(P＜0.01),而癌旁组织中CAS蛋白表达较未发生肿瘤的肝硬化、肝炎组织显著增强(P＜0.01).低分化型肿瘤细胞的CAS蛋白表达明显高于中分化型和高分化型(P＜0.01).CAS蛋白在HBV感染的肝细胞癌组织中的表达明显高于非HBV感染的癌组织(P＜0.01),其中HBV DNA阳性的肝细胞癌组织中CAS蛋白表达水平显著高于HBV DNA阴性的肝细胞癌组织(P＜0.05).结论 CAS蛋白在肝细胞癌组织中呈高表达,肝细胞癌分化愈低其表达愈强,表明CAS蛋白可作为肝细胞癌的病理诊断与分化程度的评价标志物.并推测HBV DNA可能通过上调CAS的表达,在HBV感染相关性肝癌的发生发展过程中发挥重要的作用.     

Distribution of beta 1, alpha 1, alpha 2 and alpha 3 integrin chains in basal cell carcinomas

The integrins are alpha beta heterodimeric transmembrane proteins mediating cell-substratum as well as cell-cell interactions. Changes in their expression and/or function seem to occur in a number of malignant epithelial neoplasms and may in part explain their abnormal patterns of growth and differentiation. Using monoclonal antibodies to the beta 1 (DH12), alpha 1 (TS2/7), alpha 2 (B1.515), and alpha 3 (E1.56) integrin chains, the alpha 1 beta 1 (VLA-1), alpha 2 beta 1 (VLA-2), and alpha 3 beta 1 (VLA-3) integrin receptors were studied on cryostat sections of 22 basal cell carcinomas (BCCs) and adjacent normal tissues by a standard peroxidase-antiperoxidase technique. In non-neoplastic skin, VLA-2 and VLA-3 were found in the basal layer, eccrine glands, and cells of the outer root sheath in which VLA-1 was detected. In BCCs, there was a considerably higher expression of VLA-2 and VLA-3 compared with epidermal basal cells but similar to that seen in hair bulb and outer root sheath. In two cases of nodular BCC showing evidence of regression, both VLA-2 and VLA-3 were completely negative, in contrast to non-regressing foci which were strongly positive. The high level of expression of two adhesion molecules (VLA-2 and VLA-3) involved in cell-substratum as well as cell-cell interactions may account for the more indolent pattern of growth characteristic of BCC and perhaps reflect its high degree of differentiation towards the hair follicle.     

Immunohistochemical assessment of fibronectin and tenascin and their integrin receptors alpha5beta1 and alpha9beta1 in gastric and colorectal cancers with lymph node and liver metastases

The aim of the current study was to assess immunohistochemically and compare the level of expression of tenascin (TN) and fibronectin (FN) and their integrin receptors alpha9beta1 and alpha5beta1 in the primary colorectal and gastric tumors, and in corresponding lymph node and liver metastases from 53 patients. We detected similar high deposition of the studied ECM proteins and their receptors in the stroma of primary tumors and in liver metastases and a lower deposition in lymph node metastases. Cytoplasmic immune reaction for FN and TN was also seen in the tumor cells. A pronounced co-localization of immune deposits for FN and TN and their receptors was found in the stroma of the center and the invasion front (IF) (p<0.0001). A significant decrease of FN immune signal was observed in the IF in primary tumors and liver metastases (p<0.0001). The levels of immunolabeling of FN and TN correlated with the differentiation grade of primary tumors (p<0.0001). In conclusion, we may say that there is heterogeneous deposition of TN, FN and their integrin receptors in the different areas of primary colorectal and gastric tumors and of their metastases. These findings imply that the studied proteins may be involved in cell processes such as growth, adhesion, migration and apoptosis.     

Cdc25a在跨种属肝癌组织中的表达及其意义*

 目的：研究细胞周期通路中的细胞分裂周期蛋白25a (Cdc25a)在人、大鼠和树鼩不同种属的肝癌、癌旁及正常肝组织中的表达和意义,进一步验证跨种属筛选肝癌关键基因研究策略的可行性。方法：采用实时荧光定量PCR和Western blotting技术检测人、大鼠和树鼩的肝癌及其相应癌旁组织以及正常肝组织中Cdc25a mRNA和蛋白表达水平。结果：Cdc25a mRNA表达水平在人、大鼠和树鼩的肝癌组织中均高于正常肝组织（均P<0.05）;在人和大鼠肝癌组织中的表达高于其对应的癌旁组织（P<0.05）;其余各组织间mRNA表达水平的差别无统计学意义（均P>0.05）。Cdc25a mRNA在人肝癌组织中的表达水平与病人的临床分期、门脉癌栓及肝外转移明显相关,而与术后复发、肿瘤直径、肿瘤个数、血清甲胎蛋白水平和肿瘤分化无明显关系。Western blotting检测结果显示Cdc25a在人、大鼠和树鼩肝癌组织中的蛋白表达水平均较癌旁和正常肝组织显著上调。结论：Cdc25a有可能是影响肝癌发生和发展的重要分子之一,跨种属筛选肿瘤关键基因策略可能有助于发现肝癌关键基因。     

Expression of the alpha 5 beta 1 fibronectin receptor on T lymphocytes of patients with HIV-1 infection.

AIMS: To evaluate the expression of the alpha 5 beta 1 integrin fibronectin receptor (FNR), which mediates several processes, including phagocytosis, cell motility and the immune response, on T lymphocytes of patients with HIV-1 infection. METHODS: T lymphocytes were incubated with monoclonal antibody directed against FNR and then with monoclonal antibodies, conjugated with phycoerythrin, directed against CD3, CD4 and CD8 positive cells. Expression of FNR on CD3, CD4 and CD8 positive cells was analysed using flow cytometry. RESULTS: Normal expression of FNR was observed on CD3 positive cells from asymptomatic HIV positive patients and those with AIDS. Increased expression of FNR was observed on CD8 positive cells from asymptomatic HIV positive patients and on CD4 positive cells from patients with AIDS. Increased FNR expression was observed on CD4 positive cells from patients with AIDS, particularly those with opportunistic infections caused by Pneumocystis carinii, Mycobacterium sp, Toxoplasma gondii, and Cryptococcus neoformans. CONCLUSION: Increased expression of FNR on CD8 and CD4 positive cells in asymptomatic HIV positive patients and those with AIDS, respectively, may be an epiphenomenon correlated with lymphocyte activation by HIV-1 or opportunistic infection, Further study is required to determine whether upregulation of FNR expression has a direct role in the pathogenesis of AIDS.     

Expression of integrin alpha5 and integrin beta4 and their extracellular ligands fibronectin and laminin in human decidua during early pregnancy and its sex steroid-mediated regulation

The reorganization of the human endometrium is termed decidualization, which includes endometrial cell proliferation, differentiation, integrin switching and extracellular matrix (ECM) remodeling during early pregnancy. The present study aimed to investigate distribution patterns, staining intensity and sex steroid-mediated regulation of integrin alpha5 (CD49e), integrin beta4 (CD49f) expression and their ligands fibronectin and laminin during decidualization. Human tissue samples were evaluated in two groups, those collected in early days and those collected in advanced days of the first trimester. Correlating immunostaining was found between laminin and integrin beta4, and between fibronectin and integrin alpha5. The expression of fibronectin was higher than that of laminin in the early days (p < 0.05). Temporal and spatial immunostaining of integrin beta4 and alpha5 in the apical pole of luminal and glandular cells was observed as pregnancy progressed (p < 0.05). In vitro results showed that human chorionic gonadotropin (hCG) stimulated laminin expression, downregulated integrin beta4 expression, whereas estradiol decreased fibronectin expression by Ishikawa cells. hCG suppressed fibronectin expression in endometrial stromal cells in culture. Our results suggest that fibronectin is responsible for induction of decidual cell differentiation, and different temporal and spatial expression of the integrins may play a role in implantation. Our in vitro results suggest that regulation of extracellular matrix remodeling and integrin switching are at least partially regulated by reproductive hormones.     

TLR2和TLR4在原发性肝癌中的表达

目的：本研究通过检测原发性肝癌Toll样受体2（TLR2）和Toll样受体4（TLR4）的表达，分析其与临床病理生理因素的关系，探讨TLR2，TLR4在原发性肝癌疾病过程中的可能作用。方法：采用免疫组化法和实时荧光定量PCR分别在蛋白水平及mRNA水平检测原发性肝癌组织和配对癌旁组织TLR2、TLR4的表达。结果：原发性肝癌组织在蛋白水平和mRNA水平TLR2和TLR4的表达均明显低于癌旁组织（P<0.01）。但免疫组化结果显示不论是肝癌组织还是癌旁组织，TLR2和TLR4的表达均明显高于正常肝组织（P<0.01，P<0.05），有门静脉分支癌栓的患者癌组织TLR4的表达强度低于无门静脉分支癌栓的患者（P<0.05）。结论：肝癌组织TLR2和TLR4的表达与癌旁组织相比受到相对抑制，但高于正常肝组织，TLR2和TLR4信号途径可能参与了原发性肝癌的病理过程。