Research ArticleResearch Article

In Vitro Study of Ultrasound on Multidrug Resistance in MDR Human Hepatoma HepG2 Cells

Qiujun Qi, Baojin Zhai, Yumian Guo, Zhihong Wang and Feng Wu
Chinese Journal of Clinical Oncology June 2008, 5 (3) 165-171; DOI: https://doi.org/10.1007/s11805-008-0165-5

Article Figures & Data

Figures

  • Fig.1.
    Fig.1. (A, B and C) Target and ß-actin RT-PCR mRNA levels of mdr1, MRP and LRP.

    Total RNA (0.20 mg) extracted from parental, resistant sublines and US treated resistant sublines and mdr1 ß-actin (0.8 umol), MRP ß-actin and LRP ß-actin (0.8 μmol) were used in the PCR reactions. After 32 amplification cycles, the products were resolved on a 1.5% agarose gel and stained with ethidium bromide. Lane M, a 100-bp DNA ladder as a size marker; A, HepG2/ADM treated by LIPUS; B, HepG2/ADM; C, parental cell-HepG2.

  • Fig.2.
    Fig.2. Quantification of PCR. The ratio between the mdrl or lrp and ß-action gene is expressed as described in Materials and Methods.

    A, parental cell-HepG2/WT; B, HepG2/ADM; C, HepG2/ADM treated with low intensity pulse ultrasound.

  • Fig.3.
    Fig.3. Expression of resistance-associated markers on parental, resistant sublines and treated resistant sublines.

    P-gp, MRP or LRP expression was determined by flow cytometry using monoclonal antibodies MRK16, MRPr1 and LRP-56, respectively. The fluorescence of 5,000 cells was measured at an excitation wavelength of 488 nm. The vertical and horizontal axes indicate the relative cell number and fluorescence intensity. Expression of P-gp, MRP or LRP (A, B and C), left-HepG2 cells (control: a1, b1, c1); Right-HepG2/ADM cells (a2, b2, c2); middle-HepG2/ADM cells treated with US (a3, b3, c3).

  • Fig.4.
    Fig.4. Intracellular DNR concentrations in cells treated with low intensity ultrasound.

    A, HepG2/WT treated with DNR; B, HepG2/ADM treated with DNR; C, HepG2/ADM treated with US+DNR; D. HepG2/ADM treated with VER + DNR.

Tables

  • Table 1.

    Effect of US on cellular drug sensitivity (Embedded Image).

    IC50* (mg/L)RI**
    CellsDNRADMDNRADM
    HepG20.002 ± 0.00060.003 ± 0.00081.01.0
    HepG2/ADM0.202 ± 0.00760.315 ± 0.0254101105
    HepG2/ADM + US0.072 ± 0.00020.097 ± 0.000936.031.3
    HepG2/ADM + VER0.084 ± 0.00070.094 ± 0.00654231.3
    HepG2/ADM + VER + US0.012 ± 0.00110.027 ± 0.001469

    • ↵* mean standard deviation of at least three experiments;

    • ↵** mean of three experiments; RI, resistance index. All values were calculated as described in MATERIALS AND METHODS. The concentration of verapamil (VER) was15.7 μM and the treatment time was 4 h.

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