CAJ | 학술논문

目的：克隆人长链非编码 RNA H19基因，构建表达载体，研究 H19表达对 MCF-7增殖的影响作用。方法制备乳腺癌 MCF-7细胞总 RNA，RT-PCR 扩增长链非编码 RNA H19全长序列，分子克隆至 pcDNA3．1（－）真核表达载体；分别转染 HEK-293T 和 COS 7细胞并行 Real-time qPCR 验证载体构建是否成功；转染 H19表达载体入 MCF-7细胞株，转染0、24和48 h 后行 MTS 检测 H19表达，同时设计 siRNA 小分子片段干扰 H19的表达，观察对 MCF-7细胞增殖活性的影响。结果成功克隆和构建了 hH19-pcDNA3.1（－）表达载体；转染入 MCF-7细胞48h 后，H19过表达，MTS 检测 H19表达载体组吸光度明显高于空载体组和空白对照组；而转染H19 siRNA 小分子片段后能干扰其表达，同时抑制细胞增殖。结论H19过表达能够促进乳腺癌 MCF-7细胞增殖。
목적：극륭인장련비편마 RNA H19기인，구건표체재체，연구 H19표체대 MCF-7증식적영향작용。방법제비유선암 MCF-7세포총 RNA，RT-PCR 확증장련비편마 RNA H19전장서렬，분자극륭지 pcDNA3．1（－）진핵표체재체；분별전염 HEK-293T 화 COS 7세포병행 Real-time qPCR 험증재체구건시부성공；전염 H19표체재체입 MCF-7세포주，전염0、24화48 h 후행 MTS 검측 H19표체，동시설계 siRNA 소분자편단간우 H19적표체，관찰대 MCF-7세포증식활성적영향。결과성공극륭화구건료 hH19-pcDNA3.1（－）표체재체；전염입 MCF-7세포48h 후，H19과표체，MTS 검측 H19표체재체조흡광도명현고우공재체조화공백대조조；이전염H19 siRNA 소분자편단후능간우기표체，동시억제세포증식。결론H19과표체능구촉진유선암 MCF-7세포증식。
Aims To construct an expression vector of human lncRNA H 1 9 ,and to determine the effect of H1 9 overexpression on MCF-7 cell proliferation. Methods Total RNA was extracted from MCF-7 cells,and the full-length of H1 9 lncRNA was amplified by RT-PCR and subcloned into pcDNA3.1 (-)ex-pression vector.The constructed H1 9 expression vector was transfected into HEK-293T and COS-7 cells and the H1 9 lncRNA expression was evaluated by real-time PCR.Following the transfection of H1 9 expression vec-tor into MCF-7 cells for 0,24h and 48h and H1 9 siR-NA interference fragment into MCF-7 cells for 24h, MCF-7 cell proliferation was determined by MTS as-say.Results A hH1 9-pcDNA3.1 (-)expression vector was successfully constructed. At Forty-eight hours after the transfection with H1 9 expression vector in to MCF-7 cells,cell proliferation was significantly increased in the transfected group compared to those without transfection and to those transfected with a neg-ative control vector,while twenty-four hours after the transfection with H1 9 siRNA interference fragment into MCF-7 cells,cell proliferation was significantly de-creased in the transfected group compared to those transfected with a negative control vector.Conclusion Ectopic overexpression of H1 9 lncRNA can promote breast cancer MCF-7 cell proliferation.