补中益气汤含药血清联合siRNA对肺腺癌A549/DDP细胞的LRP表达影响

Effect of Center-Supplementing and Qi-Boosting Decoction combined siRNA on LRP expression of A549/DDP cell

摘要:

目的：运用小干扰RNA片段（small interfering RNA，siRNA）的方法特异性沉默肺耐药蛋白（lung resistance protein，LRP）基因，观测该方法同补中益气汤含药血清联合应用逆转人肺腺癌耐药细胞A549/DDP的效果。方法：设计并合成针对LRP基因对应序列的siRNA，采用转染试剂对细胞进行转染。实验分组为：空白对照组、补中益气汤含药血清组、siRNA组及补中益气汤含药血清+siRNA组，利用逆转录聚合酶链反应（reverse transcription-polymerase chain reaction，RT-PCR）检测对该基因的抑制作用，Western blot及免疫细胞化学法检测蛋白质表达。结果：参照空白对照组，补中益气汤含药血清组、siRNA组及补中益气汤含药血清+siRNA组mRNA及LRP蛋白表达均减少，差异有统计学意义（P<0.05）。结论：补中益气汤含药血清和siRNA均能降低A549/DDP的LRP蛋白表达，并且2种因素之间存在交互作用。

Abstract:

Objective：To discuss the effect of concomitant application of small interfering RNA（siRNA） after targeted resection of lung resistance protein（LRP） and Buzhongyiqitang serum on the human lung adenocarcinoma drug-resistant cell-line A549/DDP. Methods：The siRNAs which had the same sequences of LRP gene were designed and synthesized，then the human lung adenocarcinoma drug-resistant cell-line A549/DPP was transected with transfection reagent. The experimental cells were grouped into control group，Buzhongyiqitang serum group，siRNA-processed group and Buzhongyiqitang plus siRNA-processed group. RT-PCR was applied to de－termine the mRNA expression of LRP. Western blot and immunocytochemical method were adopted to determine the protein expres－sion of LRP. Results：Protein and mRNA expression of LRP in Buzhongxiqitang serum group，siRNA group and Buzhongyiqitang plus siRNA-processed group was decreased significantly compared with that in control group（P<0.05）. Conclusion：Both Buzhongyiqitang serum and siRNA serve effectively in decreasing protein expression of LRP，and Buzhongyiqitang serum combined siRNA show syner－gistic effect.