摘要
目的观察核转录子κB(NF-κB)decoy寡核苷酸(ODNs)转染对结肠癌细胞NF-κB DNA结合活性的影响。方法将体外培养的人结肠癌SW480细胞随机分为5组。对照组:不作处理;脂多糖(LPS)组:LPS刺激3 h;NODN组:LPS刺激3 h后转染NF-κB decoy ODNs 6 h;SODN组:LPS刺激3 h后转染Scrambled ODNs 6 h;Lipo-fectamine2000组:LPS刺激3 h后加入同等剂量Lipofectamine2000 6 h。之后提取各组细胞核蛋白,用Western blot法检测NF-κB p65,用TransAMTM法检测细胞核NF-κB p65的DNA结合活性(用吸光度值,即A值表示)。结果细胞核中NF-κB p65的相对含量在对照组为10.1%±3.2%、LPS组为91.3%±17.8%、NODN组为86.5%±13.4%、SODN组为90.2%±14.9%、Lipofectamine2000组为89.7%±15.1%;LPS组、NODN组、SODN组、Lipo-fectamine2000组与对照组相比,P均<0.01。细胞核NF-κB p65的DNA结合活性(A值)在对照组为0.124±0.210、LPS组为1.547±0.110、NODN组为0.327±0.053、SODN组为1.509±0.172、Lipofectamine2000组为1.497±0.167;LPS组、NODN组、SODN组、Lipofectamine2000组与对照组相比,P均<0.01;NODN组与LPS组、SODN组、Lipofectamine2000组相比,P均<0.01;SODN组与Lipofectamine2000组相比,P>0.05。结论用NF-κB decoy寡核苷酸干预,可以明显抑制结肠癌细胞NF-κB p65的DNA结合活性。
Objective To observe the influence of the nuclear transcription factor kappa B （NF-KB） decoy oligode- oxynucleotides （ODNs） transfection on NF-KB DNA binding activity of colon cancer cells. Methods The cultured human colon cancer SW480 ceils in vitro were randomly divided into five groups. Control group： no treatment; the lipopolysaccha- ride （LPS） group： LPS stimulated for 3 h; NODN groups： LPS stimulated for 3 h, transfected NF-KB decoy ODNs for 6 h; SODN group ： LPS stimulated for 3 h, transfected Scrambled ODNs for 6 h ; Lipofectamine2000 group ： LPS stimulated for 3 h, added an equal dose Lipofectamine2000 for 6 h. After extracted the nuclear protein in each group, the NF-KB p65 ucle- ar protein was tested by Western blot method, the NF-KB p65 DNA binding activity was tested by TransAMTM method （A value）. Results The relative content of NF-KB p65 in the nucleus, in control group was 10.1% ± 3.2%, in LPS group was 91.3% -± 17. 8%, in NODN group was 86.5% ± 13.4%, in SODN group was 90. 2% ± 14. 9%, in Lipo- fcctamine2000 group was 89.7% ~ 15.1%. LPS group, NODN group, SODN group, Lipofectamine2000 group compared with the control group, all P 〈 0.01. The nucleus of NF-KB p65 DNA binding activity （ A value） in control group was O. 124 ±0.210, in LPS group was 1. 547 ±0. 110, in NODN group was 0. 327 ±0. 053,in SODN group was 1. 509± 0. 172, in Lipofectamine2000 group was 1. 497 ~ O. 167. LPS group, NODN group, SODN group, Lipofectamine2000 group compared with control group, all P 〈 0.01 ; NODN group compared with LPS group, SODN group, and Lipofectamine2000 group, all P 〈0.01. SODN group compared with Lipofectamine2000 group, P 〉 0.05. Conclusion NF-KB decoy oligode- oxynucleotides intervention can significantly inhibit colon cancer cell of NF-KB p65 DNA binding activity. Key words： nuclear factor KB ; oligodeoxynucleotides ; SW480 cells
出处
《山东医药》
CAS
2012年第14期4-6,共3页
Shandong Medical Journal
基金
河北省卫生厅医学科学研究重点课题基金资助项目(07063)
