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Nm23-H1核内定位对人肺腺癌A549细胞增殖的影响
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Nm23-H1核内定位对人肺腺癌A549细胞增殖的影响
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Journal Article
Nm23-H1核内定位对人肺腺癌A549细胞增殖的影响
2017
Overview
背景与目的 现有研究发现Nm23-H1还存在胞核表达,而既往的研究都是以过表达或抑制胞浆Nm23-H1为研究手段,由于Nm23-H1本身缺乏核引导序列,其研究结果并不能真实反映或重复临床中Nm23-H1以胞核定位为主的实际生物学效应。因此,本研究通过构建带有核引导序列的Nm23-H1载体并转染A549细胞以探讨Nm23-H1从胞浆向胞核转位对肺癌细胞增殖的影响。方法 采用基因重组技术构建带核定位信号序列的p Lentis-CMV-NME1-IRES2-PURO慢病毒载体,酶切和测序鉴定正确后,稳定转染A549细胞后用Western blot和激光共聚焦检测Nm23-H1蛋白的定位和表达,用CCK-8法检测细胞的增殖,流式细胞术检测细胞周期变化。结果 成功构建了核内定向表达Nm23-H1的慢病毒载体。转染组在72 h、96 h和120 h时增殖率与空载体组相比均显著升高（P〈0.000,1）。空载体组A549细胞在G0期/G1期所占比例为35.69%,高于转染组的28.28%（t=1.461,P=0.217）;而转染组细胞在G2期/M期所占比例为58.7%,空载体组为31.30%（t=4.560,P=0.010）。结论 Nm23-H1在人肺腺癌A549细胞的核内过表达使细胞主要分布在G2期/M期并促进了细胞的体外增殖。
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