中国病理生理杂志2011，Vol.27Issue(12)：2265-2269,5.DOI:10.3969/j.issn.1000-4718.2011.12.005
靶向MRP1基因的shRNA稳定逆转肺癌的多药耐药性
Stable reversal of multidrug resistance in A549/DDP cells by shRNA targeting MRP1 gene
摘要
Abstract
ADM: To reverse multidrug resistance ( MDR ) of A549/DDP cells with short hairpin RNA ( shR-NA ) expression vectors. METHODS: Two multidrug resistance - associated protein l( MRP1 ) gene - specific shRNA expression plasmids pSilencer 2. 1 - U6 neo - MRP1 were constructed and introduced into A549/DDP cells. MRP1 mRNA was assayed by real - time fluorescent quantitative PCR. The MRP1 function was determined by rhodamine 123( Rhol23 ) retention and the protein expression of MRP1 was detected by immunofluorescent staining. The viability of A549/DDP cells was evaluated by MTT method. RESULTS: MRP1 shRNA expression plasmids were successfully constructed. The expression of MRP1 at mRNA and protein levels was significantly decreased after sh - MRP1 -2. 1 - 1 and sh - MRP1 -2. 1 -2 were transfected into A549/DDP cells. The intracellular accumulation of Rhol23 significantly increased from( 16. 93 ± 0. 58 )% to ( 89. 02 ± 0. 59 )% and ( 82. 56 ± 1. 37 )%. IC50 of cisplatin were decreased from ( 101. 45 ± 0. 64 ) μmol/L to ( 38. 06 ±0. 05 ) μmol/L and ( 53. 72 ± 0. 36 ) μmol/L. lCm of 5 - fluorouracil were decreased from ( 263. 20 ± 2. 00 ) μmol/L to ( 98. 82 ± 1. 16 )μmol/L and ( 141. 81 ±0. 49 ) μmol/L. CONCLUSION: The shRNA expression plasmid pSilencer 2.1- U6 neo - MRP1 can stably and permanently inhibit MRP1 gene. The sensitivity of A549/DDP cells to drug is reversed.关键词
短发夹RNA/多药耐药/多药耐药相关蛋白1/RNA干扰/A549/DDP细胞Key words
Short hairpin RNA/ Multidrug resistance/ Multidrug resistance-associated protein 1/ RNA interference/ A549/DDP cells分类
医药卫生引用本文复制引用
邵淑丽,崔婷婷,贾红双,谢振丽,张伟伟,刘迁,陈薇薇,李爽,陈丽..靶向MRP1基因的shRNA稳定逆转肺癌的多药耐药性[J].中国病理生理杂志,2011,27(12):2265-2269,5.基金项目
黑龙江省自然科学基金资助项目(No.C200624) （No.C200624）
黑龙江省教育厅科学技术项目(No.11511447 （）
No.12511611) （）
