ESTABLISHMENT OF A NEW HUMAN CANCER CELL-LINE SECRETING PROTEASE NEXIN-II AMYLOID BETA-PROTEIN PRECURSOR DERIVED FROM SQUAMOUS-CELL CARCINOMA OF LUNG
ESTABLISHMENT OF A NEW HUMAN CANCER CELL-LINE SECRETING PROTEASE NEXIN-II AMYLOID BETA-PROTEIN PRECURSOR DERIVED FROM SQUAMOUS-CELL CARCINOMA OF LUNG
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DOI：
10.1002/ijc.2910490322
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发表时间：
1991-09-30
影响因子：
6.4
通讯作者：
KOONO, M
中科院分区：
文献类型：
--
作者：
ITOH, H;KATAOKA, H;KOONO, M
从手术切除的原发性肺癌标本中建立了一个新的人肺鳞癌细胞系（LC-1/sq）。 在无血清培养基中连续繁殖后，它分泌胰蛋白酶抑制剂到条件培养基中。 胰蛋白酶抑制剂（TI-1）的主要部分通过阴离子交换和凝胶过滤高效液相色谱（HPLC）和十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）纯化至表观均一性，然后通过transblotting至Immoblastin。 TI-1对胰蛋白酶有明显的抑制作用。 糜蛋白酶，纤溶酶和激肽释放酶的抑制程度较小，但尿激酶型纤溶酶原激活剂，弹性蛋白酶，凝血酶和木瓜蛋白酶没有受到抑制。 TI-1具有耐酸、耐热的活性，SDS-PAGE测得其分子量为115 kDa。 它具有单一的NH 2-末端序列，其NH 2-末端的前20个氨基酸残基与蛋白酶连接蛋白-II（PN-II）/淀粉样β蛋白前体（APP）的氨基酸残基相同。 LC-1/sq是第一个在体外分泌功能活性胰蛋白酶抑制剂PN-II/APP的肺鳞癌细胞系，为研究其在恶性肿瘤中的生物学意义奠定了基础。
A new cell line (LC-1/sq) of human lung squamous-cell carcinoma was established from a surgically resected specimen of primary lung cancer. Upon continuous propagation in serum-free culture medium, it secreted trypsin inhibitors into the conditioned medium. The major fraction of the trypsin inhibitor (TI-1) was purified to apparent homogeneity by anion-exchange and gel-filtration high-performance liquid chromatography (HPLC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by transblotting to Immobilion. TI-1 effectively inhibited trypsin. Chymotrypsin, plasmin and kallikrein were inhibited to a lesser extent, but urokinase-type plasminogen activator, elastase, thrombin and papain were not inhibited. The activity of TI-1 was acid-stable and heat-resistant, and its molecular weight was 115 kDa by SDS-PAGE. It exhibited single NH2-terminal sequence, and its first 20 NH2-terminal amino-acid residues were identical with those of protease nexin-II (PN-II)/amyloid beta-protein precursor (APP). These characteristics of TI-1 suggest that the major trypsin inhibitor secreted by LC-1/sq is indistinguishable from PN-II/APP. LC-1/sq is the first lung squamous carcinoma cell line that secretes functionally active trypsin inhibitor, PN-II/APP, in vitro and is useful for studying its biological significance in malignant tumor.