通过RNA干扰抑制人肝癌细胞中的MDR1基因来逆转多药耐药性。
Reversing multidrug resistance by RNA interference through the suppression of MDR1 gene in human hepatoma cells.
作者信息
Chen Xiao-Ping, Wang Qi, Guan Jian, Huang Zhi-Yong, Zhang Wan-Guang, Zhang Bi-Xiang
机构信息
Department of Surgery and Hepatic Surgery Center, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.
出版信息
World J Gastroenterol. 2006 Jun 7;12(21):3332-7. doi: 10.3748/wjg.v12.i21.3332.
AIM
To reverse the multidrug resistance (MDR) by RNA interference (RNAi)-mediated MDR1 suppression in hepatoma cells.
METHODS
For reversing MDR by RNAi technology, two different short hairpin RNAs (shRNAs) were designed and constructed into pGenSil-1 plasmid, respectively. They were then transfected into a highly adriamycin-resistant HepG2 hepatoma cell line (HepG2/ADM). The RNAi effect on MDR was evaluated by real-time PCR, cell cytotoxicity assay and rhodamine 123 (Rh123) efflux assy.
RESULTS
The stably-transfected clones showed various degrees of reversal of MDR phenotype. Surprisingly, the MDR phenotype was completely reversed in two transfected clones.
CONCLUSION
MDR can be reversed by the shRNA-mediated MDRI suppression in HepG2/ADM cells, which provides a valuable clue to make multidrug-resistant hepatoma cells sensitive to anti-cancer drugs.
目的
通过RNA干扰（RNAi）介导的多药耐药基因1（MDR1）抑制来逆转肝癌细胞中的多药耐药（MDR）。
方法
为利用RNAi技术逆转多药耐药，设计了两种不同的短发夹RNA（shRNA），并分别构建到pGenSil-1质粒中。然后将它们转染到高度耐阿霉素的HepG2肝癌细胞系（HepG2/ADM）中。通过实时定量聚合酶链反应（PCR）、细胞毒性试验和罗丹明123（Rh123）外排试验评估RNAi对多药耐药的影响。
结果
稳定转染的克隆显示出不同程度的多药耐药表型逆转。令人惊讶的是，在两个转染克隆中多药耐药表型完全逆转。
结论
shRNA介导的MDR1抑制可逆转HepG2/ADM细胞中的多药耐药，这为使多药耐药肝癌细胞对抗癌药物敏感提供了有价值的线索。
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