Inhibition of P-glycoprotein expression and reversal of drug resistance of human hepatoma HepG2 cells by multidrug resistance gene (mdr1) antisense RNA.
The development of multiple drug resistance in tumor cells is a significant problem in cancer therapy. In human, one of the reasons causing the resistance is due to the overexpression of the mdr1 gene product, P-glycoprotein. In our study, we had developed multiple drug resistant HepG2 cell line (HepG2/DR). To reverse the resistance, HepG2-DR cells were treated with antisense RNA against mdr1 gene. Total RNA and protein were extracted from the transfected cells. Northern analysis showed that mRNA level of mdr1 was decreased whereas a reduction in P-glycoprotein was detected by Western blot. By using flow cytometry, the ability of intracellular doxorubicin retention increased and drug efflux decreased in the treated cells. The result also showed that the cellular sensitivity to doxorubicin, vincristine and methotrexate measured in IC50 increased 83.3% 84.6% and 50% respectively. All these findings suggested that the expression of p-glycoprotein was successfully inhibited by antisense RNA and the drug resistance was reduced.
Chan
J Y
JY
Department of Biochemistry, The Chinese University of Hong Kong, Shatin, NT.
Chu
A C
AC
Fung
K P
KP
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000187:drug effects / Q000235:genetics*; D000970:Antineoplastic Agents / Q000493:pharmacokinetics / Q000633:toxicity*; D006528:Carcinoma, Hepatocellular; D002470:Cell Survival / Q000187:drug effects; D003001:Cloning, Molecular; D004317:Doxorubicin / Q000493:pharmacokinetics / Q000633:toxicity; D018432:Drug Resistance, Multiple / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D008113:Liver Neoplasms; D008727:Methotrexate / Q000493:pharmacokinetics / Q000633:toxicity; D016372:RNA, Antisense / Q000235:genetics / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection / Q000379:methods; D014407:Tumor Cells, Cultured; D014750:Vincristine / Q000493:pharmacokinetics / Q000633:toxicity
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000970:Antineoplastic Agents; D016372:RNA, Antisense; D014750:Vincristine; D004317:Doxorubicin; D008727:Methotrexate