摘要
目的比较补中益气汤含药血清与siRNA技术(RNA)对人肺腺癌耐顺铂细胞A549/DDP的耐药效果及对多药耐药蛋白P-gp表达的影响。方法制备补中益气汤含药血清,设计并合成针对P-gp基因对应序列的siRNA,转染A549/DDP细胞;实验分组为空白对照组、补中益气汤含药血清处理组、siRNA处理组,利用RT-PCR检测P-gp mRNA,Western印迹及免疫细胞化学法检测P-gp蛋白质的表达;MTT法检测A549/DDP细胞耐药逆转效果。结果补中益气汤含药血清和siRNA均能降低P-gp mRNA和蛋白质表达,恢复顺铂敏感性,耐药倍数分别降至2.01、2.73,具有统计学意义(P<0.05)。结论补中益气汤含药血清和siRNA均能有效逆转人肺腺癌耐药细胞A549/DDP的多药耐药。
Objective To make a comparative analysis of the effect of drug-resistance on the human lung adenocarcinoma drug-re-sistant cell-line A549/DDP between Center-Supplementing and Qi-Boosting Decoction (CQD) serum and small interfering RNA (siRNA) technology , and to discuss their effect on the expression on multi-drug resistance protein P-glycoprotein ( P-gp).Methods The serum con-taining CQD was prepared , and synthesized siRNA with sequence was designed corresponding to that of P -gp, then had the A549/DDP trans-fected.The cells were divided into control , CQD and siRNA-processed groups.Reverse transcription-polymerase chain reaction ( RT-PCR) was applied to determine the mRNA expressions of P-gp mRNA, Western blot and immunocytochemistry were adopted to determine the pro-tein expression of P-gp, and MTT was used to evaluate the effect of resistance reversal on A 549/DDP.Results Both CQD serum and siRNA significantly inhibited the mRNA and protein expressions of P-gp and restored sensitivity to cis-platinum with the resistance indexes reducing to 2.01 and 2.73 respectively ( P<0.05 ).Conclusions Both CQD and siRNA serve are effectively to reverse the multi-drug resistance of the human lung adenocarcinoma drug-resistant cell-line A549/DDP.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2014年第9期2473-2475,共3页
Chinese Journal of Gerontology
基金
国家自然科学基金资助项目(No.81072743)
辽宁省教育厅资助项目(L2010355)
关键词
补中益气汤
SIRNA
A549/DDP细胞
P-GP
耐药逆转
Center-Supplementing and Qi-Boosting Decoction
siRNA
A549/DDP cell
P-glycoprotein
Drug-resistance reversal
