Biomolecules & Therapeutics : eISSN 2005-4483 / pISSN 1976-9148

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Fig. 3. Inhibition of HAS2 expression sensitizes MCF7-DR cells to doxorubicin via reduction in NRF2 levels. (A) DR cells were transfected with non-specific control RNA (siCTRL) or CD44-specific siRNA (siCD44), and CD44 transcript levels were monitored by relative quantification real-time PCR analysis. HPRT1 was used as a housekeeping control gene. Data represent ratios with respect to siCTRL. (B) CD44, NRF2, and HAS2 protein levels were determined in siCTRL and CD44-specific siRNA-transfected siCD44 DR cells. (C, D) siCTRL and siCD44 DR cells were incubated with HA (50 ug/mL) for 24 h, and NRF2 and AKR1C1 (C) or p62 (D) levels were measured by western blotting. (E) DR cells were transfected with non-specific control RNA (siCTRL) or HAS2-specific siRNA (siHAS2) and HAS2 transcript levels were monitored by relative quantification real-time PCR analysis. HPRT1 was used as a housekeeping control gene. Data represent ratios with respect to siCTRL and are reported as the mean ± SD of two experiments. (F) CD44 protein levels were determined in siCTRL and siHAS2 DR cells using western blotting analysis. (G) NRF2, AKR1C1, and MDR1 protein levels were determined in siCTRL and siHAS2 DR cells. (H) AKR1C1 and MDR1 transcript levels were monitored in siCTRL and siHAS2 DR cells by relative quantification real-time PCR analysis. HPRT1 was used as a housekeeping control gene. Data represent ratios with respect to siCTRL and are reported as the mean ± SD of three experiments. (I) Cell viability was monitored after doxorubicin incubation for 72 h in siCTRL and siHAS2 DR cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Values represent the mean ± SD from four sampled wells. ap<0.05 compared with siCTRL. In the case of western blot results, similar blots were obtained in at least three experiments.
Biomolecules & Therapeutics 2022;30:368~379 https://doi.org/10.4062/biomolther.2022.074
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