Erbin表达缺失诱导MDA-453人乳腺癌细胞对曲妥珠单抗耐药

Deficiency of Erbin in MDA-453 human breast cancer cells induces trastuzumab resistance in vitro

目的探索Erbin表达缺失导致Her2阳性的人乳腺癌细胞MDA-453对曲妥珠单抗敏感性的影响。方法设计、构建含人Erbin基因特异性的短发夹RNA(shRNA)及对照shRNA的干扰载体,转染MDA-453人乳腺癌细胞。经G418加压筛选,获得稳定敲低Erbin的MDA-453细胞(MDA-453-Erbin sh)和稳定表达对照shRNA的MDA-453细胞(MDA-453-NC),以后者作为对照细胞。应用Western印迹法检测MDA-453-NC和MDA-453-Erbin sh细胞中Erbin表达水平。分别通过细胞增殖活性实验及细胞集落形成实验,观察敲低Erbin的MDA-453细胞对曲妥珠单抗敏感性的变化。通过免疫组化染色法,分析Erbin在人乳腺癌及正常乳腺组织中的表达。结果建立了稳定敲低Erbin的MDA-453细胞株,发现敲低Erbin表达可诱导MDA-453细胞对曲妥珠单抗产生抗性。与正常乳腺上皮组织相比,人乳腺癌组织标本中Erbin表达水平降低。结论 Erbin表达缺失可能影响乳腺癌对曲妥珠单抗治疗的敏感性。

Objective To explore the effect of Erbin deficiency in MDA-453 cells on trastuzumab（Herceptin） resist-ance .Methods The specific short hairpin RNA （ shRNA） targeting Erbin was designed and cloned into plasmid pSuppres-sor, which was subsequently transfected into MDA-453 human breast cancer cells .After being selected by G418, MDA-453 cells stably expressing Erbin shRNA were obtained and nominated as MDA-453-Erbin sh.The MDA-453 cells express-ing control shRNA （ MDA-453-NC） were used as control cells .The expression of Erbin at the protein level in MDA-453-NC and MDA-453-Erbin sh cells was analyzed by Western blotting .Cell proliferation and colony formation assays were em-ployed to investigate the effect of Erbin knockdown on the sensitivity of MDA -453 cells to trastuzumab in vitro.The levels of Erbin expression in human breast cancer tissue and normal breast tissue samples were evaluated by immunohistochemistry . Results MDA-453 cells, in which Erbin expression was stably knocked-down, were established.Deficiency of Erbin in MDA-453 cells could antagonize the anti-proliferation effects of trastuzumab in vitro.The level of Erbin expression was de-creased in some breast cancer tissue samples compared with normal breast tissue samples .Conclusion Erbin deficiency may induce the resistance of breast cancer cells to trastuzumab therapy .