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Inhibition growth of multidrug resistant KBV200 cells by MDR1 antisense RNA
Abstract:
Acquisition of resistance to multiple drugs of tumor cell caused by overexpression of the MDR1 gene is one of major obstacles in cancer chemotherapy. We have attempted to reverse the multidrug resistance (MDR) phenotype by treating vincristine (VCR) and adriamycin (ADM) resistant KBV200 cells with MDR1 antisense RNA. Retroviral vector expressing the antisense RNA was transfected into KBV200. In the transfected cells, a stable expression of antisense RNA and a reduction of cellular MDR1 mRNA could be detected by RT-PCR, and a reduction of MDR1 specific P-glycoprotein (P-gp) was also detected by Western blot, whereas an increase of the drug concentration in the cells was detected by FACS. The IC50 of transfected cells to VCR and ADM was reduced by 65 and 47%. This study demonstrates that antisense RNA can increase the sensitivity of tumor cells to anticancer drug by decreasing the expression of the MDR1 gene. This strategy may be applicable to cure cancer patients with P-gp mediated MDR phenotype.
Insights
Antisense RNA targeting the MDR1 gene reduced drug resistance in cancer cells. This approach decreased P-glycoprotein expression, increasing sensitivity to chemotherapy drugs like vincristine and adriamycin.
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Therapy
Background:
- Multidrug resistance (MDR) in tumor cells, often due to MDR1 gene overexpression, is a significant challenge in cancer chemotherapy.
- P-glycoprotein (P-gp) is a key mediator of the MDR phenotype, pumping drugs out of cancer cells.
Purpose of the Study:
- To investigate the potential of MDR1 antisense RNA to reverse the multidrug resistance phenotype in cancer cells.
- To assess the impact of MDR1 gene silencing on drug sensitivity and P-gp expression.
Main Methods:
- Transfection of KBV200 cells (resistant to vincristine and adriamycin) with a retroviral vector expressing MDR1 antisense RNA.
- Detection of antisense RNA expression, MDR1 mRNA reduction (RT-PCR), P-gp reduction (Western blot), and intracellular drug concentration (FACS).
Main Results:
- Stable expression of antisense RNA was achieved in transfected cells.
- Significant reduction in MDR1 mRNA and P-glycoprotein levels was observed.
- Increased intracellular concentrations of vincristine and adriamycin were detected.
- The IC50 values for vincristine and adriamycin decreased by 65% and 47%, respectively.
Conclusions:
- MDR1 antisense RNA effectively reduces P-gp expression, thereby increasing tumor cell sensitivity to anticancer drugs.
- This strategy holds promise for treating cancer patients with P-gp-mediated multidrug resistance.