| Cyclin D1基因沉默诱发肾癌ACHN细胞增殖受抑的研究 |
| |
| 引用本文： | 王江,郭风劲,陈安民,袁林,杨彩虹.Cyclin D1基因沉默诱发肾癌ACHN细胞增殖受抑的研究[J].华中科技大学学报(医学版),2010,39(2). |
| |
| 作者姓名： | 王江 郭风劲 陈安民 袁林 杨彩虹 |
| |
| 作者单位： | 华中科技大学同济医学院附属同济医院骨科,武汉,430030 |
| |
| 基金项目： | 国家重点基础研究发展计划资助项目 |
| |
| 摘 要： | 目的 证实Cyclin D1基因作为肿瘤基因治疗新靶点的可行性及发卡环RNA(shRNA)表达载体介导该基因沉默的有效性.方法 设计合成Cyclin D1-shRNA模板片段,与Pgenesil-1-U6载体连接,构建shRNA真核表达载体;将Pgenesil-Cyclin D1-shRNA转入ACHN肾癌细胞株.RT-PCR、Western blot分析Cyclin D1基因mRNA及蛋白表达;MTT比色法绘制细胞生长曲线;流式细胞术测细胞凋亡率;Transwell小室体外侵袭实验检测细胞侵袭.结果 Pgenesil-Cyclin D1-shRNA构建成功.Cyclin D1-shRNA实验组 Cyclin D1 mRNA抑制效果显著(0.10±0.04),明显高于Pgenesil-NC阴性对照组(0.92±0.03)及ACHN空白对照组(0.94±0.04)(均P<0.05).同样,实验组Cyclin D1蛋白表达明显下调.流式细胞术分析提示,Cyclin D1-shRNA组细胞早期及晚期凋亡细胞均较Pgenesil-NC组和ACHN组显著增高.生长曲线显示Cyclin D1-shRNA组细胞生长明显缓慢(P<0.05).Transwell实验同样发现该组细胞侵袭能力显著下降(P<0.05).结论 Cyclin D1可作为肾癌ACHN细胞生物学行为的评判标准.经载体介导shRNA干扰的高效性为肾癌基因治疗提供了实验依据.
|
| 关 键 词： | 肾ACHN细胞 RNA干扰 细胞周期蛋白D1 增殖 |
Inhibition of Proliferation Induced by Cyclin D1 Gene Silence in Human ACHN Cells |
| |
| Wang Jiang,Guo Fengjin,Chen Anmin et al.Inhibition of Proliferation Induced by Cyclin D1 Gene Silence in Human ACHN Cells[J].Journal of Huazhong University of Science and Technology(Health Sciences),2010,39(2). |
| |
| Authors: | Wang Jiang Guo Fengjin Chen Anmin |
| |
| Institution: | Wang Jiang,Guo Fengjin,Chen Anmin et al Department of Orthopedics,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China |
| |
| Abstract: | Objective To confirm feasibility of Cyclin D1 gene as a new target for cancer gene therapy and to verify the effectiveness of shRNA expression vector-mediated gene silencing.Methods A RNA interference DNA template targeting Cyclin D1 gene was designed and synthesized.By ligation,the fragment was inserted into Pgenesil-1-U6 to construct the recombinant plasmid Pgenesil-Cyclin D1-shRNA.The identified recombinant plasmid was transfected into ACHN cells with lipofactamine.Cyclin D1 mRNA and protein expression w... |
| |
| Keywords: | renal ACHN cells RNA interference Cyclin D1 proliferation |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
