Expression of aquaporin3 in human neoplastic tissues

Niu D, Kondo T, Nakazawa T, Yamane T, Mochizuki K, Kawasaki T, Matsuzaki T, Takata K & Katoh R
(2012) Histopathology  61, 543–551 Expression of aquaporin3 in human neoplastic tissues Aims: Aquaporin3 (AQP3) is distributed widely in mammalian tissues and plays an important role in fluid homeostasis. The aim of this study was to investigate the pattern of expression of AQP3 in a variety of human neoplastic tissues and to explore its diagnostic implications. Methods and results: We studied 798 neoplastic tissues using immunohistochemistry with anti‐AQP3 antibody. We demonstrated a high positive frequency of AQP3 immunoreactivity in pituitary adenomas, salivary gland tumours, thymic tumours, adenocarcinoma of the lung and prostate, squamous cell carcinomas of the skin, oesophagus and uterine cervix, apocrine carcinoma of the breast, germinal cell tumours of the ovary and testis and urothelial carcinoma of the bladder. None of the sarcomas or central nervous system tumours showed AQP3 immunoreactivity. Most tumours with a high frequency of AQP3 positivity had corresponding or surrounding normal cells that also expressed AQP3. AQP3 was not a specific marker for benign or malignant epithelial neoplasms. Conclusion: AQP3 protein is expressed in a variety of epithelial tumours limiting its use as a diagnostic marker. Furthermore, AQP3 expression in tumour cells reflected the expression status of AQP3 in the corresponding normal cells. Our data suggest that water metabolism through AQP3 is maintained during neoplastic transformation in most human tissues.     

MADD在肺腺癌组织中的表达及对肺腺癌A549细胞增殖和凋亡的影响

目的:探讨MADD在肺正常组织及肺腺癌组织中的表达及其对肺腺癌细胞增殖和凋亡的影响。方法:收集肺临床病理组织标本,免疫组化法检测肺正常组织和肺癌组织MADD表达;培养人肺腺癌A549细胞,逆转录PCR检测其IG20基因表达;用携带MADD基因的质粒和能够沉默MADD表达的慢病毒载体分别转染A549细胞,Western blot、MTT分析和流式细胞术检测其MADD表达、增殖及凋亡。结果:肺腺癌组织MADD表达水平明显高于肺正常组织和肺鳞癌组织;A549细胞能够表达MADD;高表达MADD能抑制A549细胞凋亡,提高其增殖活力,而沉默MADD表达则能促进A549细胞凋亡,降低其增殖活力。结论:肺腺癌组织MADD表达明显增高;MADD可通过抑制凋亡来促进肺腺癌细胞生存。     

Immunohistochemical study of lung adenocarcinoma using monoclonal antibody for 60-kilodalton antigen in type II pneumocytes and nonciliated bronchiolar epithelial cells. Comparison with two antisurfactant apoprotein antibodies

Monoclonal antibody KP16D3 was produced by immunizing mice with monkey bronchoalveolar lavage. KP16D3 revealed the immunohistochemical reactivity in the cytoplasm of some nonciliated bronchiolar epithelial cells and type II pneumocytes and thereby recognized specifically a protein with an apparent molecular weight of 60 kD with the use of Western blotting and immunoaffinity column chromatography followed by SDS-PAGE. Examination of 76 primary and 4 metastatic lung carcinomas in primary lung carcinoma KP16D3 showed immunohistochemical positivity only to mucin-nonproducing papillary adenocarcinoma (27/28) and bronchioloalveolar carcinoma (2/2), except for one case of large cell carcinoma. All other primary lung carcinomas such as squamous cell carcinoma, acinar adenocarcinoma, and small cell carcinoma had negative results. From these findings, KP16D3 seems to be an effective immunohistochemical marker of mucin-nonproducing papillary adenocarcinoma and bronchioloalveolar carcinoma of the lung and it appears to be useful to investigate both the histogenesis and functional expression of primary lung adenocarcinoma.     

Expression of keratinocyte growth factor/fibroblast growth factor-7 and its receptor in human lung cancer: correlation with tumour proliferative activity and patient prognosis

Keratinocyte growth factor (KGF)/fibroblast growth factor-7 (FGF-7), a mesenchymal cell-derived paracrine growth factor that specifically stimulates epithelial cell proliferation, has been implicated in the repair of lung tissue. The present study was designed to determine the expression and role of KGF and its receptor (KGFR) in human lung cancer tissues, particularly in relation to cancer cell kinetics and prognosis. Thirty-one adenocarcinomas and 30 squamous cell carcinomas, and ten normal lung tissues as a control, were examined. The expression of KGF and KGFR proteins was examined using rabbit polyclonal anti-human KGF and anti-human KGFR antisera raised in the authors' laboratories against synthetic peptides corresponding to parts of human KGF KGFR, respectively. Their specificity was confirmed in lung tumour tissues by western blotting various controls. Proliferative activity was assessed by determining the labelling index (LI) for Ki-67 antigen. Immunohistochemistry revealed that tissue co-expression of KGF KGFR correlated significantly with higher differentiation grades in squamous cell carcinoma. Conversely, in adenocarcinoma, co-expression correlated with lower differentiation grades high Ki-67 LI, was significantly associated with lymph node metastasis shorter 5-year survival. Therefore, the results indicate that co-expression of KGF KGFR correlates significantly with poor prognosis in adenocarcinoma, but not in squamous cell carcinoma, of the lung.     

水通道蛋白3在人胎盘和胎膜中的表达及分布

目的研究正常妊娠晚期人胎盘和胎膜水通道蛋白3(AQP3)的表达及分布。方法收集5例正常足月妊娠剖宫分娩的胎盘和胎膜样本,用RT-PCR测定AQP3 mRNA在胎盘和胎膜组织中的表达;用Western印迹和免疫组织化学方法检测AQP3蛋白质水平在胎盘和胎膜的表达。结果RT-PCR显示AQP3 mRNA在胎盘和胎膜组织均有表达。Western印迹结果显示胎盘组织在29 000左右有一特异性条带。免疫组织化学结果显示AQP3表达于合体滋养细胞,而在羊膜上皮细胞未见表达。结论AQP3在胎盘母儿液体平衡中可能发挥重要作用。     

Immunohistochemical Demonstration of Nonspecific Cross-reacting Antigen in Normal and Neoplastic Human Tissues Using a Monoclonal Antibody: Cornparison with Carcinoembryonic Antigen Localization

Nonspecific cross-reacting antigen (NCA) immunoreactivity was localized in normal and neoplastic human tissues using a monoclonal antibody to 55, 90 and 95 kDa molecules of NCA. This was compared to the localization of immunoreactive carcinoembryonic antigen (CEA) as demonstrated by polyclonal and monoclonal antibodies. In frozen sections, CEA was localized in normal surface epithelium of the stomach and colon where NCA was only weakly detected. Type 1 and type 2 like pneumocytes were positive for NCA, while CEA was localized only in type 2-like pneumocytes. CEA and NCA were both demonstrated in ductal cells of frozen pancreatobiliary and mammary tissues. The antigenicity of CEA and NCA in normal tissues was significantly lost after paraffin embedding as compared to frozen sections. NCA was consistently demonstrated in eccrine sweat glands embedded in paraffin. In various tumor tissues, CEA and NCA were colocalized and expression increased sufficiently to be detected in paraffin sections. Adenocarcinomas of the stomach and colon and cystadenocarcinoma of the pancreas, as well as neuroendocrine carcinomas of the lung and thyroid, showed a CEA predominance over NCA. In ductal adenocarcinomas of the pancreas and breast and in cholangiocarcinoma, NCA reactivity was greater than CEA. Keratiniring foci of most squamous cell carcinomas of mucosal origin and some adenocarcinomas equally expressed both. Hepatocellular carcinoma, lobular mammary carcinoma and papillary thyroid carcinoma were positive only with unabsorbed polyclonal antibody which widely recognizes CEA-related substances. Renal cell carcinoma, prostatic adenocarcinoma, transitional cell carcinoma, anaplastic carcinomas, choriocarcinoma and basal cell carcinomas showed little or no immunoreactivity. Hence the relative ratio of CEA/NCA expression in tumors was dependent on the tissue of origin and histologic type. The cytoplasmic granular staining of NCA in cancer cells was a noteworthy difference from the plasma membrane-associated localization of CEA. Acta Pathol Jpn 40: 85–97, 1990.     

Immunohistochemical demonstration of nonspecific cross-reacting antigen in normal and neoplastic human tissues using a monoclonal antibody. Comparison with carcinoembryonic antigen localization

Nonspecific cross-reacting antigen (NCA) immunoreactivity was localized in normal and neoplastic human tissues using a monoclonal antibody to 55, 90 and 95 kDa molecules of NCA. This was compared to the localization of immunoreactive carcinoembryonic antigen (CEA) as demonstrated by polyclonal and monoclonal antibodies. In frozen sections, CEA was localized in normal surface epithelium of the stomach and colon where NCA was only weakly detected. Type 1 and type 2-like pneumocytes were positive for NCA, while CEA was localized only in type 2-like pneumocytes. CEA and NCA were both demonstrated in ductal cells of frozen pancreatobiliary and mammary tissues. The antigenicity of CEA and NCA in normal tissues was significantly lost after paraffin embedding as compared to frozen sections. NCA was consistently demonstrated in eccrine sweat glands embedded in paraffin. In various tumor tissues, CEA and NCA were colocalized and expression increased sufficiently to be detected in paraffin sections. Adenocarcinomas of the stomach and colon and cystadenocarcinoma of the pancreas, as well as neuroendocrine carcinomas of the lung and thyroid, showed a CEA predominance over NCA. In ductal adenocarcinomas of the pancreas and breast and in cholangiocarcinoma, NCA reactivity was greater than CEA. Keratinizing foci of most squamous cell carcinomas of mucosal origin and some adenocarcinomas equally expressed both. Hepatocellular carcinoma, lobular mammary carcinoma and papillary thyroid carcinoma were positive only with unabsorbed polyclonal antibody which widely recognizes CEA-related substances. Renal cell carcinoma, prostatic adenocarcinoma, transitional cell carcinoma, anaplastic carcinomas, choriocarcinoma and basal cell carcinomas showed little or no immunoreactivity. Hence the relative ratio of CEA/NCA expression in tumors was dependent on the tissue of origin and histologic type. The cytoplasmic granular staining of NCA in cancer cells was a noteworthy difference from the plasma membrane-associated localization of CEA.     

肺鳞状细胞癌中EGFR和ERK2共表达与ERK2激活的关系

目的:从蛋白和mRNA水平研究EGFR和ERK2在肺鳞状细胞癌(SCC)中的共表达及其与ERK2激活之间的关系。方法:分别采用Western blotting和反转录PCR(RT-PCR)对20例SCC及其配对正常肺组织中的EGFR和ERK2蛋白和mRNA的表达进行检测,ERK2的活性采用免疫沉淀和激酶反应进行测定。结果:在配对的正常肺组织中EGFR和ERK2的mRNA和蛋白的表达几乎检测不到或仅有少量表达,但在20例肺癌组织中表达显著上调,其中肺癌组织中ERK2蛋白的表达比正常肺组织上调3-5倍;肺癌组织中ERK2的活性也比正常肺组织显著增加。结论:在SCC中,EGFR和ERK2的蛋白共表达发生了上调,主要归因于其基因转录水平的激活,并可能最终导致肺癌组织中ERK2活性的增加。     

水通道蛋白3在人胎盘和胎膜中的表达及分布

目的 研究正常妊娠晚期人胎盘和胎膜水通道蛋白3(AQP3)的表达及分布.方法 收集5例正常足月妊娠剖宫分娩的胎盘和胎膜样本,用RT-PCR测定AQP3 mRNA在胎盘和胎膜组织中的表达;用Western印迹和免疫组织化学方法检测AQP3蛋白质水平在胎盘和胎膜的表达.结果 RT-PCR显示AQP3 mRNA在胎盘和胎膜组织均有表达.Western印迹结果显示胎盘组织在29 000左右有一特异性条带.免疫组织化学结果显示AQP3表达于合体滋养细胞,而在羊膜上皮细胞未见表达.结论 AQP3在胎盘母儿液体平衡中可能发挥重要作用.     

Pathological study of carbohydrate antigens in human lung cancer with monoclonal antibodies]

The expressions of carbohydrate antigens were examined with panel of specific anti-carbohydrate monoclonal antibodies (MAbs) on human lung cancer tissues. The MAbs used were SH1, SH2, SNH3, AH6, CA3F4, TKH2, TKH6 and TKH5 which define Lex, dimeric Lex, sialosyl Lex, Ley, Lea, sialosyl Tn, Tn and fucosyl GM1, respectively. Paraffin-embedded tissue sections (30 squamous cell carcinomas, 30 adenocarcinomas and 27 small cell carcinomas) were tested by immunohistological staining. Evaluation of stained specimens was performed by taking mean value of scores evaluated by three independent examiners. In squamous cell carcinoma, expression of Ley, sialosyl Tn, sialosyl Lex and Lea was significantly higher than other antigens. Lex was also expressed especially in keratinized tissues. In adenocarcinoma, Lea was expressed most remarkably. Sialosyl Lex, Ley, sialosyl Tn were also highly expressed in malignant cells. There was no significant difference in staining patterns between well and poorly differentiated adenocarcinomas. Sialosyl Tn and sialosyl Lex were positive in morphologically normal mucous glands adjacent to tumors. In small cell carcinomas, Ley was expressed more than other types of tumors, whereas Lex and sialosyl Tn were less than others. Tn antigen was observed throughout adenocarcinomas, squamous and small cell carcinomas in a relatively weak manner. Dimeric Lex and fucosyl GM1 antigens were not detected. Most of normal lung sections showed negative staining with those MAbs. These findings indicate that there are differential expressions of certain carbohydrate antigens in different types of human lung cancers based on their origins.