The reversion effect of the RNAi-silencing mdr1 gene on multidrug resistance of the leukemia cell HT9.
Overexpression of P-glycoprotein (P-gp), the mdr1 gene product, confers multidrug resistance (MDR) to tumor cells and often limits the efficacy of chemotherapy. This study evaluated RNAi for specific silencing of the mdr1 gene and reversion of multidrug resistance. Three different short hairpin RNAs (shRNAs) were designed and constructed in a pSilencer 3.1-H1 neo plasmid. The shRNA recombinant plasmids were transfected into HT9 leukemia cells. The RNAi effect was evaluated by real-time PCR, Western blotting and cell cytotoxicity assay. In the cell, shRNAs can specifically down-regulate the expression of mdr1, mRNA and P-gp. Resistance against harringtonine, doxorubicin and curcumin was decreased. The study indicated that shRNA recombinant plasmids could modulate MDR in vitro.
Shao
Shu-Li
SL
College of Life Science, Beijing Normal University, 19 Xinjiekouwai Street, Beijing 100875, China. shaoshuli2008@163.com
Sun
Ying-Yu
YY
Li
Xu-Yan
XY
Zhang
Wei-Wei
WW
Fu
Bo
B
Yun
Dong-Ze
DZ
Zuo
Ming-Xue
MX
D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D005822:Genetic Vectors; D018922:HL-60 Cells; D006801:Humans; D007938:Leukemia / Q000378:metabolism* / Q000473:pathology; D034622:RNA Interference; D014162:Transfection
C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1