A novel antisense oligodeoxynucleotide-doxorubicin conjugate against the multidrug resistance gene mdr1:inhibition to KB-A-1 cell and P-glycoprotein expression

Abstract

Abstract: AIM: Over-expression of P-glycoprotein (P-gp) on the surface of tumor cells that works as a membrane pump enhancing the drug efflux is regarded as a main candidate mechanism of multidrug resistance (MDR).And mdr1 is one of the two main genes that encode P-gp in human cells.In this study, a novel conjugate made of mdr1-anti-sense oligodeoxynucleotide (ODN) and a potent anticancer drug doxorubicin was constructed.The cytotoxicity of the conjugate as well as the effect of the conjugate on the modulation of P-gp-mediated MDR in KB-A-1 cell lines was studied.The molecular mechanism of its regulation effect was also investigated. METHODS: Multidrug resistant KB-A-1 cell lines were used.MTT was applied to measure the cytotoxicity of the conjugate and doxorubicin to KB-A-1 cells. Effect of the conjugate on intracellular doxorubicin accumulation was determined by HPLC.RT-PCR and Western blot were used to determine the expression of mdr1 gene and P-glycoprotein in KB-A-1 cells.RESULTS: The results showed that the conjugate exhibited more cytotoxicity than ODN, and also it could promote the cytotoxicity of doxorubicin to KB-A-1 cells.The IC50 of doxorubicin dropped from 24 to 18 μg·ml^-1.The results of RT-PCR and Western blot analysis illuminated that conjugate might down-regulate the P-gp expression.CONCLUSION: All the findings suggest that the conjugate is more potential for efficient reversal of MDR phenomenon than free ODN. The study indicated that such a conjugate should be a new feasible and efficient anti-sense reagent both in lab research and further in clinical therapy.

Key words: oligodeoxynucleotide, conjugate, cytotoxicity, multidrug resistance, P-glycoprotein

CLC Number:

ZHAN Xiao-Yun, REN Yu-Hong, ZHANG Qiang, LU Yan-Hua, WEI Dong-Zhi, LIU Jian-Wen. A novel antisense oligodeoxynucleotide-doxorubicin conjugate against the multidrug resistance gene mdr1:inhibition to KB-A-1 cell and P-glycoprotein expression[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2004, 9(4): 403-410.

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