CAJ | 학술논문

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。
목적：탐색Erbin표체결실도치Her2양성적인유선암세포MDA-453대곡타주단항민감성적영향。방법설계、구건함인Erbin기인특이성적단발협RNA（ shRNA）급대조shRNA적간우재체，전염MDA-453인유선암세포。경G418가압사선，획득은정고저Erbin적MDA-453세포（ MDA-453-Erbin sh）화은정표체대조shRNA적MDA-453세포（ MDA-453-NC），이후자작위대조세포。응용Western인적법검측MDA-453-NC화MDA-453-Erbin sh세포중Erbin표체수평。분별통과세포증식활성실험급세포집락형성실험，관찰고저Erbin적MDA-453세포대곡타주단항민감성적변화。통과면역조화염색법，분석Erbin재인유선암급정상유선조직중적표체。결과건립료은정고저Erbin적MDA-453세포주，발현고저Erbin표체가유도MDA-453세포대곡타주단항산생항성。여정상유선상피조직상비，인유선암조직표본중Erbin표체수평강저。결론 Erbin표체결실가능영향유선암대곡타주단항치료적민감성。
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 .