敲低E2F1基因对肾透明细胞癌细胞株Caki-2细胞增殖和侵袭力的影响  

Effect of E2F1 Knockdown on proliferation and invasion of clear cell renal cell carcinoma cell lineCaki -2

作　　者：高宇[1] 范阳[1] 陈伟浩[1] 黄庆波[1] 艾青[1] 倪栋[1] 马鑫[1] 张旭[1] 

机构地区：[1]解放军总医院泌尿外科,北京100853

出　　处：《中华实验外科杂志》2013年第2期333-335,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30972982）;国家高技术研究发展计划（863计划）资助项目（2012AA02101）

摘　　要：目的观察敲低E2F1基因后对肾透明细胞癌细胞株Caki-2细胞增殖、迁移和侵袭的影响。方法选取肾透明细胞癌细胞株Caki-2，应用脂质体lip02000将E2F1小干扰RNA（siRNA）序列和阴性对照E2F1序列转染进细胞株，采用实时定量聚合酶链反应（Real—timePCR）和Westernblot法检测转染后E2F1的表达，并通过MTS和Transwell法检测细胞增殖、迁移和侵袭能力的变化。结果转染siRNA—E2F1组的E2F1mRNA水平为（37．770±3．782）×10-5，比较未转染组（130．100±6．301）×10。和阴性对照组（134．600±7．163）×10。明显降低（P〈0．05），E2F1蛋白水平也明显降低。在Caki-2中敲低E2F1后24、48、72、96hMTS吸光度（A）值明显下降（P〈0．05），转染后48h细胞迁移实验中，转染siRNA—E2F1组的穿膜细胞数为（38．330±4．041），比较未转染组（83．000±8．888）和阴性对照组（87．330±6．429），穿膜细胞数明显减少（P〈0．01）；在细胞侵袭能力实验中，转染siRNA．E2F1组的穿膜细胞数为（78．330±7．572），比较未转染组（143．300±10．066）和阴性对照组（147．000±14．000），穿膜细胞数明显减少（P〈0．01）。结论在肾透明细胞癌细胞株Caki-2中敲低E2F1基因表达后，其细胞增殖、迁移和侵袭能力明显下降。Objective To study the effect of E2F1 knockdown on proliferation, migration and in- vasion of clear cell renal cell carcinoma cell line Caki-2. Methods Small interfering RNA （siRNA） -E2F1 and negative control sequences were transfected into Caki-2 cells by Lipo2000. The E2F1 expression levels were detected by using real-time quantitative polymerase chain reaction （PCR） and Western blotting re- spectively after transfection. MTS and Transwell assays were used to detect variations in cell proliferation, migration and invasion. Results Compared to untransfected group （ 130. 100 ± 6. 301 ） x 10 -5 and negative control group （134. 600 _± 7. 163 ） x 10-5, the mRNA levels in siRNA-E2F1 group （37. 770 ± 3.782） x 10-5 were significantly reduced （P 〈 0. 01 ）. The protein levels were consistent with mRNA levels. MTS assay demonstrated the absorbance in siRNA-E2F1 group was significantly decreased at the time points of 24, 48, 72 and 96 h after transfeetion （ P 〈 0. 05 ） , and Transwell assay revealed that the number of migrating cells in siRNA-E2F1 group （38. 330 ±4. 041 ） was significantly reduced at 48 h after transfection as com- pared with untransfected group （ 83. 000 ± 8. 888 ） and negative control group （ 87. 330 ± 6. 429 ） （ P 〈 O. 01 ）, and the number of invasive cells in siRNA-E2F1 group （78. 330 ±7. 572） was strongly reduced at 48 h after transfection as compared with untransfeeted group （ 143.30 ± 10. 066） and negative control group （ 147. 000 ± 14. 000） （ P 〈 0. 01 ）. Conclusion Knockdown of E2F1 significantly decreased proliferation, migration and invasion of Caki-2 cells.

关 键 词：肾透明细胞癌 E2F1基因 增殖 迁移 侵袭 

分 类 号：R737[医药卫生—肿瘤]