siRNA抑制NF-κBp65对Bcl-2及肝癌细胞凋亡的影响  

SiRNA-mediated inhibition of NF-κB p65 down-regulations Bcl-2 expression and promotes apoptosis in hepatocellular carcinoma cell lines

作　　者：赵心恺[1,2] 宁巧明[1] 孙晓宁[1] 田德安[2] 

机构地区：[1]海南省人民医院,海南省海口市570311 [2]华中科技大学同济医学院同济医院消化内科,湖北省武汉市430030

出　　处：《世界华人消化杂志》2011年第22期2358-2362,共5页World Chinese Journal of Digestology

基　　金：海南省卫生厅科研立项课题;No.琼卫2010-8~~

摘　　要：目的:研究siRNA抑制NF-κBp65后Bcl-2在肝癌细胞中的变化及对肝癌细胞凋亡的影响.方法:选择肝癌细胞HepG2、SMMC7721和人胚胎肝细胞LO2细胞株,应用Westernblot法分别检测NF-κBp65、Bcl-2在不同细胞中的表达;应用siRNA方法抑制NF-κBp65在肝癌细胞中的表达,观察NF-κBp65、Bcl-2在肝癌细胞中的变化及应用流式细胞仪观察肝癌细胞的凋亡情况.结果:Westernblot法检测显示NF-κBp65、Bcl-2在肝癌细胞HepG2、SMMC7721中的表达明显高于人胚胎肝细胞LO2(2.14±0.19,2.09±0.27vs0.54±0.11;1.42±0.15,1.47±0.14vs0.60±0.08,均P<0.05),而NF-κBp65、Bcl-2在肝癌细胞HepG2、SMMC7721中的表达差异无统计学意义;应用siRNA抑制NF-κBp65的表达后,应用Westernblot法检测显示NF-κBp65在肝癌细胞HepG2、SMMC7721中的表达较未转染组明显下降(2.08±0.19vs0.99±0.12;2.03±0.17vs0.94±0.14,均P<0.05),说明转染成功.随后应用Westernblot法检测显示Bcl-2在肝癌细胞HepG2、SMMC7721中的表达较未转染组明显下降(1.37±0.05vs0.72±0.02;1.44±0.03vs0.69±0.03,均P<0.05),且肝癌细胞凋亡较未转染组明显增加(5.12%±0.61%vs37.87%±4.10%;5.80%±0.71%vs40.19%±3.78%,均P<0.05).结论:siRNA可以成功抑制NF-κBp65在肝癌细胞中的活性,进一步证实NF-κBp65对Bcl-2具有调控作用,他们共同参与了肝癌的发生发展过程.AIM： To investigate the impact of small interfering RNA （siRNA）-mediated inhibition of nuclear factor-κB P65 （NF-κB p65） on Bcl-2 expression and apoptosis in human hepatocellular carcinoma （HCC） cell lines.METHODS： HCC cell lines HepG2, SMMC7721 and human fetal liver cell line LO2 were used in the study. The expression of NF-κB p65 and Bcl-2 in the above three cell lines was detected by Western blot. SiRNA technology was then used to inhibit NF-κB p65 to observe the effect of NF-κB p65 knockdown on Bcl-2 expression and cell apoptosis.RESULTS： The expression levels of NF-κB p65 and Bcl-2 in HepG2 and SMMC7721 cells were significantly higher than those in LO2 cells （2.14 ± 0.19, 2.09 ± 0.27 vs 0.54 ± 0.11; 1.42 ± 0.15, 1.47 ± 0.14 vs 0.60 ± 0.08, all P 〈 0.05）. No significant difference was detected in the expression levels of NF-κB p65 and Bcl-2 between HepG2 and SMMC7721 ceils. SiRNA transfection significantly down-regulated NF-κB p65 expression in HepG2 and SMMC7721 ceils compared to non- transfected cells （2.08 ± 0.19 vs 0.99 ± 0.12; 2.03 ± 0.17 vs 0.94 ± 0.14, both P 〈 0.05）. SiRNA-mediated NF-κB p65 knockdown significantly downregulated Bcl-2 expression （1.37 ± 0.05 vs 0.72 ± 0.02; 1.44 ± 0.03 vs 0.69 ± 0.03, both P 〈 0.05） and increased apoptosis （5.12% ± 0.61% vs 37.87% ± 4.10%; 5.80% ± 0.71% vs 40.19% ± 3.78%, both P 〈 0.05） in HepG2 and SMMC7721 cells compared to non-transfected ceils. CONCLUSION： SiRNA-mediated NF-κB p65 knockdown significantly down-regulates Bcl-2 expression and promotes apoptosis in HepG2 and SMMC7721 cells.

关 键 词：核因子ΚB P65 BCL-2 肝细胞癌 

分 类 号：R735.7[医药卫生—肿瘤]