补中益气汤与siRNA对肺腺癌A549/DDP细胞耐药逆转作用的比较  

Effect of drug resistance reversal of lung adnocarcinoma A540 /DDP by Center-supplementing and Qi-boosting decoction and siRNA

作　　者：李璐璐[1] 于丹[1] 王莹[1] 井欢[1] 刘春英[1] 

机构地区：[1]辽宁中医药大学基础医学院病理教研室,辽宁沈阳110032

出　　处：《中国老年学杂志》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)

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.

关 键 词：补中益气汤 SIRNA A549/DDP细胞 P-GP 耐药逆转 

分 类 号：R743.2[医药卫生—神经病学与精神病学]