S—100蛋白阳性Langerhans细胞在食管癌发生中的计量学观察

Langerhans细胞(简称LC)参与免疫反应,它在肿瘤发生过程中所起的作用尚不完全清楚。对一些肿瘤组织内LC数量的研究已有报道。而食管癌发生过程中LC数量的变化尚未见报道。本文用S-100蛋白抗体免疫组化ABC法显示并分析了食管正常鳞状上皮、不典型增生、原位癌及各级鳞状细胞癌组织中LC的数量和分布。     

食管癌中S—100蛋白阳性Langerhans细胞的形态学观察

通过抗S-100蛋白抗体ABC免疫组化方法观察分析了27例人食管鳞状细胞癌组织中S-100蛋白阳性Langerhars细胞的形态及分布，结果显示在食管癌发生中的正常粘膜上皮、不典型增生上皮、原位癌各各级鳞状细胞癌组织中均有该细胞分布，Langerhans细胞表现为分枝或未分枝两种形态，其形态随病变的不同而有所改变，癌组织中的Lanerhans细胞与癌细胞、淋巴细胞密接触，其中部分癌细胞发生退行性改变，提示S-100蛋白阳性Langerhans细胞参与机体抗肿瘤免疫反应。     

食管脱落上皮细胞中p53基因突变的检测

目的 探讨在食管拉网细胞中进行食管癌基因检测的方法及可行性,了解食管癌前期病变细胞中p53基因的突变与癌变的关系。方法 对四川省盐亭县1982年进行食管癌普查的食管拉网脱落细胞涂片标本,应用聚合酶链反应-单链构象多态性分析(PCR-SSCP)方法,检测其p53基因第5外显子及第7外显子的突变情况。结果 全组48例标本中,食管正常上皮和重度不典型增生上皮各24例,p53基因检测均获成功。食管重度不典型增生上皮细胞中有5例检测到突变,均为p53基因第7外显子突变,而第5外显子未检测到突变;正常食管鳞状上皮拉网脱落细胞中未发现突变。检测到突变的5例食管上皮重度不典型增生者,有3例分别在10年、12年和14年后转变为食管癌。结论p53基因突变的食管上皮不典型增生细胞具有明显的癌变趋势;对食管拉网脱落细胞涂片标本进行微量DNA的提取、扩增和基因检测,可作为研究食管癌前期病变的一种新方法。     

食管癌发生发展过程中环氧合酶-2蛋白表达的研究

目的 研究环氧合酶－2（Cox-2)蛋白在食管癌及癌前病变组织中的表达,探讨非类固醇抗炎药在食管癌高危人群中化学预防的可能性。方法 应用免疫组织化学方法检测120例食管癌（原位癌30例,鳞状细胞癌60例,腺癌30例）及其113例增生性病变（单纯增生29例,轻、中、重度不典型增生分别为31例、30例、23例）和27例正常食管黏膜鳞状上皮,以及3例Barrett食管组织中Cox-2蛋白的表达情况。结果 在正常食管黏膜上皮,单纯增生,轻、中、重度不典型增生,原位癌未发现有Cox-2蛋白表达;在6％（4／60）侵袭性鳞状细胞癌和70％（21／30）的食管腺癌中,Cox-2蛋白表达阳性。结论 Cox-2蛋白表达可能与食管腺癌的形成有关,而与鳞状细胞癌的发生发展无关。     

uPA、PAI-1在食管癌发生、发展不同阶段的表达及意义

目的:探讨尿激酶型纤溶酶原激活物(uPA)、纤溶酶原激活物抑制剂-1(PAI-1)在食管癌发生、发展中的作用及意义。方法:采用免疫组织化学SP法检测正常食管黏膜上皮组织(18例)、食管黏膜上皮不典型增生组织(23例)及食管癌组织(68例)中uPA、PAI-1蛋白的表达,并分析其和食管癌临床病理特征的关系。结果:uPA蛋白在正常食管黏膜上皮组织、食管黏膜上皮不典型增生组织及食管癌组织中的阳性率分别为27.8%、39.1%、70.6%,三者间有显著性差异(P<0.05);PAI-1蛋白在正常食管黏膜上皮组织、食管黏膜上皮不典型增生组织及食管癌组织中的阳性率分别为22.2%、34.8%、64.7%,三者间有显著性差异(P<0.05)。uPA、PAI-1蛋白表达与年龄、性别、病理类型无关(P>0.05),在临床病理Ⅰ-Ⅱ期和Ⅲ-Ⅳ期间有升高的趋势,但差异无统计学意义(P>0.05),与分化程度和淋巴结有无转移有关(P<0.05)。结论:uPA、PAI-1蛋白在食管癌发生、发展不同阶段的表达呈进行性上升的趋势,可能与食管癌的发生、发展相关,过度表达提示预后不良。     

膜联蛋白-1在食管鳞状细胞癌发生发展中的表达差异

目的　检测膜联蛋白 -1(annexin -1)在食管鳞状细胞癌及癌前病变中的表达差异。方法　应用免疫组化S -P方法检测 13 5例食管鳞状细胞癌及其癌旁正常鳞状上皮、原位癌及其不典型增生病变中annexin -1的表达情况。结果　annexin -1在 12 9例 (95 .6% )癌旁正常食管鳞状上皮呈强阳性表达 ;在 3 5例 (2 5 .9% )鳞状细胞癌呈阳性表达 ,60例 (4 4 .4% )为弱阳性 ,40例(2 9.6% )为阴性 ,统计学处理有极显著性差异 (P <0 .0 1)。食管鳞状细胞癌annexin -1表达下降与患者的年龄、性别、肿瘤分化程度及淋巴结转移无关 (P >0 .0 5 )。annexin -1在原位癌和不典型增生病变的表达亦呈下降趋势 ,与正常鳞状上皮比较有非常显著性差异 (P <0 .0 1)。结论　annexin -1蛋白在食管鳞状细胞癌和癌前病变中有不同程度的丢失 ,它很可能成为食管癌早期诊断以及预测食管癌前病变癌变风险的具有重要应用价值的指标     

共刺激分子B7-H3在人食管鳞癌中的表达及临床意义

目的 分析人食管鳞癌组织中B7-H3分子的表达与患者临床病理因素以及术后生存时间之间的关系。方法 采用免疫组织化学法检测82例人食管癌组织、对应癌旁正常食管组织及食管鳞状上皮不典型增生组织中B7-H3分子的表达以及CD8+T淋巴细胞的浸润程度。结果 食管癌组织中B7-H3蛋白的阳性表达率为98.8%(81/82),而B7-H3在癌旁及鳞状上皮不典型增生组织中几乎不表达。B7-H3的表达与患者年龄、肿瘤浸润深度及TNM分期相关,差异有统计学意义（P<0.05）,与肿瘤组织中CD8+T淋巴细胞浸润程度呈负相关(P<0.05),与患者预后呈负相关（P<0.05）。结论 B7-H3在食管癌中过表达,和肿瘤的进展密切相关。食管癌中B7-H3分子的表达可能参与了抑制食管癌组织内CD8+T细胞功能,促使肿瘤细胞逃避免疫监视,提示B7-H3在食管癌诊断、判断预后及靶向治疗中具有潜在的临床应用价值。     

宫颈癌的上皮膜抗原的免疫组化研究

目的 探讨上皮膜抗原表达在宫颈癌早期诊断中的价值。方法 采用免疫组化方法对宫颈正常上皮、不典型增生上皮、原位癌及浸润癌进行上皮膜抗原表达检测。结果 正常宫颈上皮及轻度、中等度上皮表层细胞呈弱阳性,重度不典型增生上皮几乎全层阳性表达,原位癌及浸润癌全层呈强阳性表达。结论 宫颈上皮的上皮膜抗原分布范围及表达强度与上皮细胞的异型性有关。对宫颈癌的早期诊断有临床意义。     

成束蛋白和膜联蛋白I在食管鳞状细胞癌癌前病变中的表达

背景与目的：研究食管鳞状细胞癌癌变早期异常改变的蛋白质以发现与食管癌早期病变相关的特征性标志分子。探讨了成束蛋白和膜联蛋白Ⅰ在食管癌癌前病变中的表达情况。方法：应用免疫组化方法分析食管癌高发现场癌前病变样本中成束蛋白和膜联蛋白Ⅰ的表达水平，用X^2检验对比成束蛋白和膜联蛋白Ⅰ在不同程度癌前病变和食管癌中的表达差异。其中包括，食管鳞状细胞癌癌前病变54例、正常食管上皮8例和中晚期食管癌9例成束蛋白的表达水平；以及食管鳞状细胞癌前病变52例、正常食管上皮11例和中晚期食管癌7例的膜联蛋白Ⅰ表达水平。结果：与正常食管鳞状上皮相比，成束蛋白在食管癌及其癌前病变中表达增强，其表达阳性率为低度癌前病变（轻度和中度不典型增生）85．7％（24／28）、高度癌前病变（重度不典型增生和原位癌）84．6％（22／26）和中晚期食管癌88．9％（8／9）。然而，膜联蛋白Ⅰ在食管癌及其癌前病变中表达降低或丢失，其表达阳性率分别为低度癌前病变14．3％（4／28）、高度癌前病变8．3％（2／24）和中晚期食管癌0％（0／7）。与正常食管上皮相比，成束蛋白和膜联蛋白Ⅰ在食管低度癌前病变中表达程度的异常均具有显著意义，P值分别为0．003和0．000。结论：成束蛋白异常增强和膜联蛋白Ⅰ的丢失与食管癌癌前病变相关。     

P33~(ING1)和Survivin及Ki67蛋白在食管癌发生发展中的作用

目的 检测P33ING1、Survivin和Ki67蛋白在食管正常黏膜、单纯增生、不典型增生、浸润癌中的表达情况,分析这三种蛋白在食管癌不同病变区域中的表达及与食管癌病理特征的关系.方法 采用免疫组化方法 检测P33ING1、Survivin和Ki67蛋白在51例食管癌大体标本中的不同病变区域的表达.结果 P33ING1蛋白从正常黏膜、单纯增生、不典型增生增生、浸润癌呈渐进性低表达,而Survivin和Ki67蛋白则呈渐进性高表达.三种蛋白在正常黏膜中的表达与不典型增生及浸润癌中的表达差异有显著性(P<0.01);P33ING1蛋白在食管癌中的表达与癌肿浸润深度(肌层至全层)呈负相关(P<0.05),与分化程度(G1、G3)呈正相关(P<0.05 );Survivin蛋白表达与食管癌分化程度、淋巴结转移有关(P<0.05);P33ING1蛋白在浸润癌中的低表达与Survivin和Ki67蛋白高表达呈负相关(P<0.01).结论 Survivin功能的激活、Ki67增殖及P33ING1功能的下调可能共同对抗细胞凋亡,参与食管癌的发生发展.三者有可能为临床上食管癌早期诊断、相关治疗及预后研究提供新的手段.     

PTEN和CyclinE蛋白在子宫内膜癌组织中的表达

目的探讨子宫内膜良性、癌前病变和恶性病变中PTEN、CyclinE蛋白的表达及其意义.方法采用免疫组化S-P法检测正常子宫内膜、子宫内膜增生、非典型性增生和子宫内膜癌组织中PTEN和CyclinE蛋白.结果非典型性增生组和子宫内膜样癌组中PTEN和CyclinE蛋白阳性表达率分别为60.0%,50.0%和42.0%,62.2%.非典型性增生组中PTEN的阳性表达显著低于正常组和增生组(P<0.02),而CyclinE的阳性表达则显著高于正常组和增生组(P<0.03).PTEN和CyclinE蛋白在子宫内膜样癌组中的表达呈负相关(r=-0.4475,P<0.01).宫内膜样癌中PTEN蛋白阳性表达的缺失与组织学分级(P<0.02)及肌层浸润深度或伴有转移有关(P<0.04),与临床分期无关(P>0.05).CyclinE的阳性表达率则与组织学分级有关(P<0.05),与肌层浸润(P>0.05)和临床分期无关(P>0.05).结论 PTEN表达缺失和CyclinE的过度表达与子宫内膜样癌的发生发展密切相关,二者的联合检测可作为子宫内膜样癌早期诊断的生物学指标.     

Adenosine triphosphatases as histochemical markers for the cell of origin in experimental mammary carcinoma.

Different adenosine triphosphatase (ATPase) activities were detected at an ultrastructural level in order to differentiate epithelial and myoepithelial cells in normal and neoplastic mouse mammary tissues. Mg2+ dependent and Na+-K+-dependent ATPase activities were studied in: BALB/c mouse mammary gland; a BALB/c carcinoma from a transplantable D2 hyperplastic nodule; a stable cell line, MCF-8, derived from the BALB/c carcinoma; and a BALB/c scirrhous-like carcinoma induced by MCF-8 cell inoculation. Mg2+-dependent ATPase was detected in the plasma membranes of the normal mouse mammary epithelial cells, the epithelial component of the BALB/c carcinoma, the MCF-8 cells in culture, and the atypical epithelial component of the scirrhous-like carcinoma. Na+-K+-dependent and Mg2+-dependent ATPase were localized in the plasma membranes of the myoepithelial cells of the normal mammary gland and the BALB/c carcinoma. The results from these histochemical studies established that the cell of origin in both the BALB/c carcinoma and the scirrhous-like carcinoma was the mammary epithelial rather than the myoepithelial cells. Furthermore, these results indicated that the MCF-8 cell line was derived from the epithelial component of the primary BALB/c carcinoma. These conclusions, which were based on histochemical study, were supported by the presence of intracisternal type A viral particles in the epithelial cells of the primary BALB/c carcinoma, the MCF-8 cells in culture, and the epithelial cells of the scirrhous-like carcinoma. Thus, the enzymatic markers were specific for cell type and remained unchanged by the process of cell transformation.     

46161例食管上皮正常、增生癌变时间、癌变率的研究

 对食管上皮细胞正常（以下简称正常）、食管上皮细胞轻度增生（以下简称轻度增生）和食管上皮细胞重度增生（以下简称重度增生）共46161例进行了癌变时间和癌变率的研究结果说明：食管癌变平均时间的频率差异为重度增生癌变平均时间3年7月＜轻度增生癌变平均时间5年4月＜食管正常癌变平均时间8年10月。食管癌变率的频率差异为重度增生癌变率38．9%＞轻度增生癌变率5．8%＞正常癌变率1．4%。     

Angiogenesis in colon hyperplastic polyp

Despite the significance of tumor angiogenesis and the extensive knowledge on the molecular basis of blood vessel formation in carcinoma of colorectum, no data exist in hyperplastic polyp. This prompted us to examine angiogenesis in hyperplastic polyp. Eleven small hyperplastic polyps, 13 large hyperplastic polyps and their adjacent normal mucosas were included in this study. Angiogenesis was assessed by immunohistochemistry using monoclonal antibody against CD34. Angiogenic factor, thymidine phosphorylase was also examined by immunohistochemistry. Intra-tumoral microvessel density (IMD) in large hyperplastic polyp was significantly higher than that in small hyperplastic polyp (P<0.01) and that in normal mucosa (P<0.01). DAD in small hyperplastic polyp was also significantly higher than that in normal mucosa (P<0.01). Expression of dThdPase was almost observed in stromal cells in normal, small and large hyperplastic polyp. In addition, the proportion of the stromal cells expressing dThdPase in large hyperplastic polyp was significantly higher than that in small hyperplastic polyp and normal tissue (P<0.01, respectively). The proportion of the stromal cells expressing dThdPase in small hyperplastic polyp was significantly higher than that in normal tissue (P<0.01). The present study provides that angiogenesis may have an important role(s) in the development of hyperplastic polyp and dThdPase in stromal cells may support angiogenesis in hyperplastic polyp. Anti-angiogenic therapy might be available for suppression of hyperplastic polyp.     

粘着斑激酶在食管癌中表达的临床意义

目的探讨粘着斑激酶蛋白(focal adhesion k inase,FAK)的表达与食管癌发生、浸润转移及预后的关系。方法应用免疫组织化学En V ision法原位观察正常食管粘膜、单纯增生粘膜上皮、各级不典型增生上皮及癌组织中FAK蛋白的表达。结果正常食管粘膜未见FAK蛋白的表达。单纯增生粘膜上皮、不典型增生上皮及癌组织中FAK的阳性表达率分别为为10.0%、31.25%及62.5%,其中原位癌的阳性表达率为41.7%,浸润癌的阳性表达率为65.2%。无淋巴结转移者的阳性表达率为22.9%,有淋巴结转移者的阳性表达率为82.6%(P=0.0002)。FAK蛋白阳性表达率随肿瘤分化程度降低及浸润深度加深而增加(P<0.05,P<0.01)。FAK阳性表达者术后的3年和5年生存率分别为41.54%和38.38%,FAK阴性表达者的3年和5年生存率分别为89.66%和72.41%,两组3年及5年生存率的差别有显著性差异(P=0.0011和P=0.026)。结论FAK蛋白的表达与食管癌的发生,细胞分化,浸润,淋巴结转移及预后有关,FAK蛋白表达阳性的患者比表达阴性的患者预后差。     

Induction of ductal and stromal hyperplasia by basic fibroblast growth factor produced by human pancreatic carcinoma

A histopathology study of 22 pancreatic adenocarcinoma cases revealed that 13 of the patients presented with hyperplastic lesions (atypical and non-atypical hyperplasia, mucous cell hypertrophy, focal epithelial hyperplasia, and ductal papillary hyperplasia), and 9 exhibited fibrosis adjacent to the carcinoma. All lesions expressed high levels of epidermal growth factor receptor (EGF-R) (p<0.0001 and p=0.0008, respectively) as compared with normal ductal epithelium. Non-atypical and atypical hyperplastic lesions also had a higher proliferating cell nuclear antigen (PCNA) labeling index (p<0.001 and p=0.0008, respectively) than normal ductal epithelium. A gradient in PCNA+ nuclei was found in acinar cells adjacent to the tumors. In 16 cases with marked fibrosis, we observed a significant increase of PCNA+ nuclei in stromal fibroblasts (p=0.0041) and significant upregulation of basic fibroblast growth factor (bFGF) mRNA expression in adjacent tumor cells (p=0.0213). These data suggest that the production of bFGF by pancreatic cancer cells induces ductal and stromal hyperplasia of the pancreas.     

食管鳞癌组织中RECK和MMP-9基因mRNA检测与分析

目的 :探讨RECK和MMP-9在食管鳞状上皮细胞癌中的作用及其与临床病理因素的关系。 方法 :收集河南省安阳市肿瘤医院食管癌标本62例、癌旁不典型增生组织31例及正常食管粘膜组织62例。应用原位杂交方法检测每例食管标本RECK和MMP-9mRNA的表达情况。应用χ²检验,Speannan等级相关分析在SPSS13.0统计软件上作相关统计。 结果 :在食管鳞癌癌变过程中RECK在食管鳞癌组织、癌旁不典型增生组织及正常食管粘膜组织中mRNA的表达率依次增高,分别为45.2%(28/62)、61.3%(19/31)、82.3%(51/62),组间比较差异有统计学意义(χ²=19.186,P<0.05);不同分化程度、不同浸润深度及有无淋巴结转移的食管鳞癌组织之间RECKmRNA阳性表达率差异均有统计学意义(χ²分别为6.799、7.862、9.121,P均<0.05)。食管鳞癌组织及癌旁不典型增生组织中MMP-9mR-NA表达均高于正常食管粘膜组织,表达率分别为71.0%(44/62)、54.8%(17/31)、48.4%(30/62),组间比较差异有统计学意义(χ²=6.876,P<0.05);食管鳞癌组织中MMP-9mRNA表达与癌组织的分化程度、不同浸润深度及有无淋巴结转移密切相关(χ²分别为7.458,11.737,3.916,P均<0.05)。RECK和MMP-9mRNA表达率均与食管癌患者的性别、年龄无显著相关关系(P均>0.05)。RECK和MMP-9的mRNA的表达呈负相关关系(γ=-0.348,P<0.01)。 结论 :RECK mRNA表达率降低与MMP-9mRNA表达率升高可能与食管鳞癌的发生、发展及浸润转移有关。     

Suppressive Effect of the Angiogenesis Inhibitor TNP-470 on the Development of Carcinogen-induced Hepatic Nodules in Rats

Tumor metastasis can be prevented by inhibiting angiogenesis. In the present study, we have demonstrated that the angiogenesis inhibitor TNP-470 also suppresses the development of primary hepatic nodules. Hepatocarcinogenesis was performed by the feeding of 2-acetylaminofluorene to hepatectomized rats during 8–14 weeks of age. Predominantly arterial-to-portal circulation and sinusoidal capillarization were determined by the staining of nodules with arterially infused ink and immunostaining for factor VIII-related antigen, respectively. Intraperitoneal administration of 30 mg/kg b.w. of TNP-470 twice a week significantly reduced the number of hepatic nodules. Among the nodules, hyperplastic nodules stained with ink, atypical hyperplastic nodules and hepatocellular carcinoma, all of which possess structurally altered sinusoidal endothelial cells or capillary-type endothelial cells, were dramatically decreased in number. Suppression was observed equally in nodules of all sizes. TNP-470 was more effective when administered during 8–20 weeks than during 14–26 weeks. In contrast, ink-non-stained hyperplastic nodules, which have normal sinusoidal endothelial cells, were not affected at all. The present results indicate that TNP-470 suppresses the development of primary hepatic nodules whose microvessels are capillaries or transitional forms from sinusoids to capillaries.     

Differentiation between Cancerous and Normal Hyperplastic Lobules in Breast Lesions

Determining the risk that a particular area of hyperplastic breast tissue will progress to cancer is difficult and is currently expressed only as a general risk factor within the population. Using an antibody against the apoptotic purinergic receptor P2X7, we examined 40 cases each of the following histological categories: normal, moderate, florid and atypical hyperplasia, lobular carcinoma in situ, ductal carcinoma in situ, invasive lobular and invasive ductal carcinoma. These were previously diagnosed by H&E and supplied by clinical laboratories as tissue sections. Normal and mildly hyperplastic epithelium was devoid of the cytolytic P2X7 receptors whereas all epithelial cells in all cases of in situ or invasive lobular or ductal carcinoma labelled intensely. The lobular and ductal in situ cases labelled intracellularly while the invasive epithelial cancer cells showed intense cell surface label indicating an attempt was being made to induce apoptosis. All these receptors however are non-functional and thus unable to induce apoptosis. Approximately 10% of all hyperplastic lobules examined in the biopsied tissue, regardless of H&E classification, labelled for P2X7, which is suggestive of early metabolic cancerous change. The acini within lobules were either completely labelled with P2X7 or were completely devoid of the receptor. A potential advantage of this method lies in identifying early cancerous change in hyperplastic lobules and in establishing the true extent of cancerous spread in infiltrating lesions, thus facilitating the task of reporting clear surgical margins.     

Behavior of cervical epithelium during carcinogenesis: autoradiography of tritiated cytidine in the C57BL mouse.

The autoradiographic patterns of the cervical epithelia of 39 virgin C57BL mice after in vivo injection of [3H]cytidine was investigated. Of the 22 animals painted with benzo[a]pyrene for 5 months, 8 developed intraepithelial atypias and 2 had invasive carcinoma. The mean number of silver grains in individual cells increased from histologically normal cervical epithelium through atypical epithelium to invasive carcinoma. It is apparent that the autoradiographic patterns in normal, atypical, and invasive carcinoma cells reflected quantitative differences in cytidine incorporation. These differences may denote increased demand for RNA precursors in the cervical epithelium during carcinogenesis.