Reversal of multi-drug resistance by vector-based-ShRNA-Mdr1 <Emphasis Type="Italic">In Vitro</Emphasis> and <Emphasis Type="Italic">In Vivo</Emphasis> |
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Authors: | Shi Lu Qi Huang Zehua Wang Yinfeng Song Lijun Wang |
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Institution: | [1]Department of Obstetrics and Gynecology, Union Hospital, Tongfi Medical College, Huazhong University of Science and Technology, Wuhan 430030, China [2]Department of Thoracic and Cardiac Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China |
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Abstract: | In order to investigate the effects of vector-based hairpin small interference RNA (shRNA) on the reversal of multi-drug resistance
(mdr) of A2780/Taxol cells, a novel vector pEGFP-H1/mdr1 containing mdr1-shRNA targeting at position 2943–2963 of mdr1 was
designed and synthesized. Subsequently, A2780/Taxol cells were transfected with pEGFP-H1/mdr1, and the expression of mdr1
mRNA and P-gp was detected by using RT-PCR and Western blot respectively. MTT was used to measure the 50% inhibition concentration
(IC50) of Taxol to A2780/Taxol cells. The results showed that at the 24th and 48th h after transfection, the expression of mdr1
mRNA was decreased to (52.1±1.0)% and (0.01±1.7)%, and that of P-gp decreased to (88.3±2.1)% and 0%, respectively. At the
48th h after transfection, the relative reversal rate of A2780/Taxol cells to Taxol was 69.54%. In vivo, the nude mice xenografts were injected with pEGFP-H1/mdr1, and then administrated Taxol. The tumor volume in pEGFP-H1/mdr1-transfected
group was significantly reduced as compared with that in blank control group or pEGFP-H1-transfected group (807.20±103.16
vs 1563.78±210.54 or 1480.78±241.24 mm3, both P<0.01). These results suggested that transfection of pEGFP-H1/mdr1 could efficiently down-regulate the expression of mdr1
mRNA and P-gp in A2780/Taxol cells, and effectively restore the sensitivity of A2780/Taxol cells to Taxol both in vitro and in vivo. |
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Keywords: | RNA interference multi-drug resistance gene therapy |
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