MDR mRNA and MDR Proteins Expression (Measured by RT-PCR and by Flow Cytometry, Respectively), Effect of Probenecid on Calcein Efflux, and Effect of CsA on Calcein-AM Uptake for 13 Cell Lines
| Cell Lines . | mRNA Expression Measured by RT/PCR* (mean ± SD)-151 . | Protein Expression Measured by Flow Cytometry-152 (mean ± SD)-151 . | Effect of Probenecid on Calcein Efflux-153 (mean ± SD)-151 . | Effect of CsA on Calcein-AM Uptake-153 (mean ± SD)-151 . | ||||
|---|---|---|---|---|---|---|---|---|
| MDR1 . | MRP . | LRP . | Pgp . | MRP . | LRP . | |||
| A549 | 0 ± 0 | 1.11 ± 0.39 | 1.25 ± 0.33 | 1 ± 0.03 | 2.88 ± 0.24 | 13.2 ± 6.7 | 1.51 ± 0.08 | 1 ± 0 |
| K562 | 0 ± 0.01 | 0.1 ± 0.03 | 0.12 ± 0.02 | 1 ± 0.06 | 1.73 ± 0.24 | 15.1 ± 3 | 1.31 ± 0.05 | 1 ± 0 |
| HHT30 | 0.64 ± 0.53 | 0.46 ± 0.17 | 0.36 ± 0.02 | 1.4 ± 0.1 | 1.86 ± 0.14 | 12.5 ± 1.6 | 1.33 ± 0.04 | 1 ± 0 |
| HHT100 | 0.93 ± 0.68 | 0.08 ± 0.07 | 1.11 ± 0.14 | 6.8 ± 1.5 | 1.65 ± 0.11 | 11 ± 2.8 | 1.23 ± 0.08 | 4.9 ± 1 |
| HHT200 | 1.32 ± 0.6 | 0.21 ± 0.1 | 0.59 ± 0.31 | 17.2 ± 2.5 | 1.56 ± 0.1 | 11 ± 3.3 | 1.17 ± 0.05 | 16.3 ± 6.1 |
| HHT300 | 2.65 ± 1.06 | 0.33 ± 0.39 | 0.9 ± 0.26 | 71.3 ± 7.2 | 1.84 ± 0.18 | 19.3 ± 4.4 | 1.25 ± 0.11 | 39 ± 7.5 |
| HHT400 | 2.3 ± 0.79 | 0.38 ± 0.1 | 0.11 ± 0.01 | 40.8 ± 8.5 | 1.89 ± 0.19 | 24.2 ± 5.3 | 1.29 ± 0.06 | 24.5 ± 4.2 |
| HL60 | 0 ± 0.01 | 0.59 ± 0.11 | 0.24 ± 0.11 | 1.1 ± 0.2 | 2.18 ± 0.28 | 29.7 ± 7.5 | 1.32 ± 0.15 | 1 ± 0.1 |
| HL60 Pgp | 8.74 ± 2.16 | 0.38 ± 0.03 | 0.32 ± 0.06 | 125.3 ± 18.9 | 1.49 ± 0.22 | 13.8 ± 3.5 | 1.2 ± 0.05 | 46.7 ± 9.9 |
| HL60 MRP | 0.4 ± 0.51 | 2.14 ± 0.24 | 0.02 ± 0.04 | 2.5 ± 0.6 | 5 ± 2.4 | 1.1 ± 0.3 | 1.85 ± 0.13 | 10.9 ± 8.1 |
| CEM | 0.5 ± 0.24 | 0.21 ± 0.05 | 0.01 ± 0.01 | 2.2 ± 0.5 | 1.58 ± 0.4 | 1.2 ± 0.4 | 1.28 ± 0.04 | 1 ± 0.2 |
| CEM/VLB | 3.21 ± 0.97 | 0.26 ± 0.05 | 0.1 ± 0.02 | 52.8 ± 10.4 | 1.65 ± 0.21 | 1.3 ± 0.2 | 1.27 ± 0.06 | 18.5 ± 4.5 |
| U937 | 0 ± 0.02 | 0 ± 0.03 | 0.74 ± 0.27 | 1.2 ± 0.4 | 1.07 ± 0.13 | 12.6 ± 5.9 | 1.17 ± 0.02 | 1 ± 0.1 |
| Cell Lines . | mRNA Expression Measured by RT/PCR* (mean ± SD)-151 . | Protein Expression Measured by Flow Cytometry-152 (mean ± SD)-151 . | Effect of Probenecid on Calcein Efflux-153 (mean ± SD)-151 . | Effect of CsA on Calcein-AM Uptake-153 (mean ± SD)-151 . | ||||
|---|---|---|---|---|---|---|---|---|
| MDR1 . | MRP . | LRP . | Pgp . | MRP . | LRP . | |||
| A549 | 0 ± 0 | 1.11 ± 0.39 | 1.25 ± 0.33 | 1 ± 0.03 | 2.88 ± 0.24 | 13.2 ± 6.7 | 1.51 ± 0.08 | 1 ± 0 |
| K562 | 0 ± 0.01 | 0.1 ± 0.03 | 0.12 ± 0.02 | 1 ± 0.06 | 1.73 ± 0.24 | 15.1 ± 3 | 1.31 ± 0.05 | 1 ± 0 |
| HHT30 | 0.64 ± 0.53 | 0.46 ± 0.17 | 0.36 ± 0.02 | 1.4 ± 0.1 | 1.86 ± 0.14 | 12.5 ± 1.6 | 1.33 ± 0.04 | 1 ± 0 |
| HHT100 | 0.93 ± 0.68 | 0.08 ± 0.07 | 1.11 ± 0.14 | 6.8 ± 1.5 | 1.65 ± 0.11 | 11 ± 2.8 | 1.23 ± 0.08 | 4.9 ± 1 |
| HHT200 | 1.32 ± 0.6 | 0.21 ± 0.1 | 0.59 ± 0.31 | 17.2 ± 2.5 | 1.56 ± 0.1 | 11 ± 3.3 | 1.17 ± 0.05 | 16.3 ± 6.1 |
| HHT300 | 2.65 ± 1.06 | 0.33 ± 0.39 | 0.9 ± 0.26 | 71.3 ± 7.2 | 1.84 ± 0.18 | 19.3 ± 4.4 | 1.25 ± 0.11 | 39 ± 7.5 |
| HHT400 | 2.3 ± 0.79 | 0.38 ± 0.1 | 0.11 ± 0.01 | 40.8 ± 8.5 | 1.89 ± 0.19 | 24.2 ± 5.3 | 1.29 ± 0.06 | 24.5 ± 4.2 |
| HL60 | 0 ± 0.01 | 0.59 ± 0.11 | 0.24 ± 0.11 | 1.1 ± 0.2 | 2.18 ± 0.28 | 29.7 ± 7.5 | 1.32 ± 0.15 | 1 ± 0.1 |
| HL60 Pgp | 8.74 ± 2.16 | 0.38 ± 0.03 | 0.32 ± 0.06 | 125.3 ± 18.9 | 1.49 ± 0.22 | 13.8 ± 3.5 | 1.2 ± 0.05 | 46.7 ± 9.9 |
| HL60 MRP | 0.4 ± 0.51 | 2.14 ± 0.24 | 0.02 ± 0.04 | 2.5 ± 0.6 | 5 ± 2.4 | 1.1 ± 0.3 | 1.85 ± 0.13 | 10.9 ± 8.1 |
| CEM | 0.5 ± 0.24 | 0.21 ± 0.05 | 0.01 ± 0.01 | 2.2 ± 0.5 | 1.58 ± 0.4 | 1.2 ± 0.4 | 1.28 ± 0.04 | 1 ± 0.2 |
| CEM/VLB | 3.21 ± 0.97 | 0.26 ± 0.05 | 0.1 ± 0.02 | 52.8 ± 10.4 | 1.65 ± 0.21 | 1.3 ± 0.2 | 1.27 ± 0.06 | 18.5 ± 4.5 |
| U937 | 0 ± 0.02 | 0 ± 0.03 | 0.74 ± 0.27 | 1.2 ± 0.4 | 1.07 ± 0.13 | 12.6 ± 5.9 | 1.17 ± 0.02 | 1 ± 0.1 |
*The results were given as the ratio of quantities of MDR mRNA product/β2m mRNA product.
The experiments were performed in triplicate, and at three different times.
Protein values were expressed as adjusted for control, ie, as ratio of arithmetic mean fluorescence of UIC2/IgG2A control or MRPm6/IgG1 control or LRP56/IgG2b control.
All the data are ratios of drug fluorescence with modulator divided by drug fluorescence without modulator after subtraction of the fluorescence of the control.