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Elevated expression of annexin II (lipocortin II p36) in a multidrug resistant small cell lung cancer cell line.

机译:Annexin II(lipocortin IIp36)在多药耐药的小细胞肺癌细胞系中的表达升高。

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摘要

The doxorubicin-selected multidrug resistant small cell lung cancer cell line, H69AR, is cross-resistant to the Vinca alkaloids and epipodophyllotoxins, but does not overexpress P-glycoprotein, a 170 kDa plasma membrane efflux pump usually associated with this type of resistance. Monoclonal antibodies were raised against the H69AR cell line and one of these, MAb 3.186, recognises a peptide epitope on a 36 kDa phosphorylated protein that is membrane associated, but not presented on the external surface of H69AR cells (Mirski & Cole, 1991). In the present study, in vitro translation and molecular cloning techniques were used to determine the relative levels of mRNA corresponding to the 3.186 antigen. In addition, a cDNA clone containing an insert of approximately 1.4 kb was obtained by screening an H69AR cDNA library with 125I-MAb 3.186. Fragments of this cloned DNA hybridised to a single mRNA species of approximately 1.6 kb that was 5 to 6-fold elevated in H69AR cells. Partial DNA sequencing and restriction endonuclease mapping revealed identity of the cloned DNA with p36, a member of the annexin/lipocortin family of Ca2+ and phospholipid binding proteins.
机译:阿霉素选择的多药耐药小细胞肺癌细胞株H69AR对长春花生物碱和表鬼臼毒素具有交叉耐药性,但不会过表达P-糖蛋白,P-糖蛋白是一种170 kDa的质膜外排泵,通常与这种耐药有关。产生了针对H69AR细胞系的单克隆抗体,其中一种抗体MAb 3.186识别与膜相关但未在H69AR细胞外表面呈递的36 kDa磷酸化蛋白上的肽表位(Mirski&Cole,1991)。在本研究中,体外翻译和分子克隆技术用于确定对应于3.186抗原的mRNA的相对水平。另外,通过用125I-MAb 3.186筛选H69AR cDNA文库,获得了含有约1.4kb插入物的cDNA克隆。此克隆的DNA片段与大约1.6 kb的单个mRNA物种杂交,在H69AR细胞中升高了5至6倍。部分DNA测序和限制性核酸内切酶作图揭示了克隆的DNA与p36的同一性,p36是Ca2 +的膜联蛋白/脂皮质激素家族成员和磷脂结合蛋白。

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