pcDNA3.1-CD74-ROS1重组质粒构建与表达对肺癌细胞增殖及迁移的影响
DOI: 10.3971/j.issn.1000-8578.2018.17.1486
Keywords: CD74-ROS1,融合PCR,非小细胞肺癌,迁移,增殖,miR-601 Suppresses Migration and Invasion of Non-small Cell lung Cancer Cells by Targeting MMP-17,Effects of Sanguinarine on Invasion, Migration and Wnt/β-catenin Signaling Pathway of Lung Adenocarcinoma Cells,Expression and Functional of LncRNA XIST in MCF-7 Breast Cancer Cells,Autophagy Related Protein 5 Regulates Colon Cancer Cell Senescence Through p53/p21 Pathway,Effect of Down-regulation of HOXB7 Expression on Proliferation, Migration and Invasion Abilities of Hepatoma Cells,Curcumin Reverses Ionizing Radiation-induced Epithelial Mesenchymal Transformation of Non-small Cell Lung Cancer A549 Cells,Effects of miR-144 on Proliferation, Migration, Invasion and PI3K Pathway of Pancreatic Cancer SW1990 Cells,Effect of ANLN Gene Silencing on Invasion, Migration, Proliferation and Apoptosis of Gastric Cancer Cells,Research Progress on Resistance Mechanisms and Overcoming Strategies to Thirdgeneration EGFR-TKIs in Advanced Non-small Cell Lung Cancer,Advances in Prognostic Value of Peripheral Blood Inflammatory Indexes in Non-small Cell Lung Cancer,Mechanism of D-bifunctional Protein Promoting Formation of Hepatocellular Carcinoma in Rat via STAT3,Regulatory Effect of Astragaloside A on Proliferation and Apoptosis of Human Colon Cancer SW480 Cell Line,Effect of FBXL20 on Proliferation, Migration and Invasion of Gastric Cancer Cells,Challenges of Immune Checkpoint Inhibitors in Treatment of Non-small Cell Lung Cancer,Downregulation of Skp2 Expression Inhibits Malignant Phenotype of Osteosarcoma U2OS Cells
Abstract:
摘要 目的 构建C-ros致癌基因1（ROS1）融合基因CD74-ROS1重组质粒，并转染A549细胞株，检测其对肺癌细胞迁移能力的影响。方法 通过PCR技术获得CD74和ROS1基因片段，利用融合PCR法构建CD74-ROS1融合基因，构建pcDNA3.1-CD74-ROS1重组质粒，通过脂质体转染技术转入A549细胞中，利用Real-time PCR和Western blot技术检测CD74-ROS1的表达。MTT法观察细胞增殖情况，结晶紫染色法观察细胞形态变化，划痕实验检测CD74-ROS1对肺癌细胞迁移能力的影响。结果 酶切及测序结果表明CD74-ROS1融合基因构建成功，阳性克隆的CD74-ROS1蛋白表达显著增加。CD74-ROS1过表达导致细胞增殖能力增强，细胞形态上发生上皮-间质样转变且细胞迁移能力增强。结论 pcDNA3.1-CD74-ROS1重组质粒构建成功，并在A549细胞中稳定表达，CD74-ROS1过表达能够促进肺癌细胞增殖、形态转变且增强其迁移能力
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