Representative AML cell line data
Cell line . | Expression, mean ± SD (no. independent assays) . | . | . | . | Efflux, mean ± SD (no. independent assays) . | . | Cytotoxicity, mean % ± SD (no. independent assays) . | . | . | . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| . | CD33 . | pBzR . | Pgp . | MRP1 . | DiOC2 + CSA . | CDCF + MK571 . | GO - untrtd . | GO + PK - PK only . | GO + CSA - CSA only . | GO + MK - MK only . | |||||||
| TF1 | 263 ± 38 (11) | 2577 ± 434 (3) | 1.5 ± 0.2 (11) | 1.9 ± 0.1 (9) | 6.1 ± 0.9 (9) | 52.9 ± 10.0 (9) | − 4 ± 1 (5) | − 20 ± 4 | − 17 ± 5 | − 2 ± 1 | |||||||
| P | — | — | — | — | — | — | < .01* | < .01† < .01‡ | NS, < .05‡ | NS, NS | |||||||
| ML-1 | 62 ± 19 (3) | 4551 ± 925 (5) | 0.8 (1) | 1.0 (1) | 1.0 ± 0 (3) | 3 ± 0 (3) | − 36 ± 2 (5) | − 31 ± 2 | − 34 ± 3 | − 24 ± 2 | |||||||
| P | — | — | — | — | — | — | < .0001* | NS, < .0001‡ | NS, < .0001‡ | NS, < .0001‡ | |||||||
| NB4 | 48 ± 8 (7) | 973 ± 157 (3) | 0.9 ± 0 (5) | 1.9 ± 0.1 (5) | 1.1 ± 0 (3) | 4.6 ± 1.1 (4) | − 15 ± 3 (4) | − 34 ± 5 | − 34 ± 1 | ± 17 ± 4 | |||||||
| P | — | — | — | — | — | — | < .05* | < .01†, < .0001‡ | < .001†, < .0001‡ | NS, NS | |||||||
| HL-60 | 106 ± 2 (29) | 3228 ± 700 (3) | 0.9 ± 0.1 (19) | 1.8 ± 0.1 (18) | 1.1 ± 0 (11) | 19.3 ± 3.5 (12) | − 26 ± 4 (5) | − 42 ± 4 | − 46 ± 1 | − 32 ± 6 | |||||||
| P | — | — | — | — | — | — | < .0001* | < .01†, < .0001‡ | < .001†, < .01‡ | NS, < .01‡ | |||||||
Cell line . | Expression, mean ± SD (no. independent assays) . | . | . | . | Efflux, mean ± SD (no. independent assays) . | . | Cytotoxicity, mean % ± SD (no. independent assays) . | . | . | . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| . | CD33 . | pBzR . | Pgp . | MRP1 . | DiOC2 + CSA . | CDCF + MK571 . | GO - untrtd . | GO + PK - PK only . | GO + CSA - CSA only . | GO + MK - MK only . | |||||||
| TF1 | 263 ± 38 (11) | 2577 ± 434 (3) | 1.5 ± 0.2 (11) | 1.9 ± 0.1 (9) | 6.1 ± 0.9 (9) | 52.9 ± 10.0 (9) | − 4 ± 1 (5) | − 20 ± 4 | − 17 ± 5 | − 2 ± 1 | |||||||
| P | — | — | — | — | — | — | < .01* | < .01† < .01‡ | NS, < .05‡ | NS, NS | |||||||
| ML-1 | 62 ± 19 (3) | 4551 ± 925 (5) | 0.8 (1) | 1.0 (1) | 1.0 ± 0 (3) | 3 ± 0 (3) | − 36 ± 2 (5) | − 31 ± 2 | − 34 ± 3 | − 24 ± 2 | |||||||
| P | — | — | — | — | — | — | < .0001* | NS, < .0001‡ | NS, < .0001‡ | NS, < .0001‡ | |||||||
| NB4 | 48 ± 8 (7) | 973 ± 157 (3) | 0.9 ± 0 (5) | 1.9 ± 0.1 (5) | 1.1 ± 0 (3) | 4.6 ± 1.1 (4) | − 15 ± 3 (4) | − 34 ± 5 | − 34 ± 1 | ± 17 ± 4 | |||||||
| P | — | — | — | — | — | — | < .05* | < .01†, < .0001‡ | < .001†, < .0001‡ | NS, NS | |||||||
| HL-60 | 106 ± 2 (29) | 3228 ± 700 (3) | 0.9 ± 0.1 (19) | 1.8 ± 0.1 (18) | 1.1 ± 0 (11) | 19.3 ± 3.5 (12) | − 26 ± 4 (5) | − 42 ± 4 | − 46 ± 1 | − 32 ± 6 | |||||||
| P | — | — | — | — | — | — | < .0001* | < .01†, < .0001‡ | < .001†, < .01‡ | NS, < .01‡ | |||||||
Expression data are specific immunostaining measures, except for pBzR expression data that represent tritiated PK 11195 binding blocked by excess unlabeled PK11195. Efflux data represent dye accumulation that was reduced by transporter-selective blockers, CSA or MK-571. Cells that were Pl− and did not show reduced DiOC6(3) staining were scored as “live” in flow cytometry toxicity assays. GO cytotoxicity is presented as specific live cell fractions, relative to untreated live cell fractions. The cytotoxicity of GO + sensitizer treatments is expressed with sensitizer effects also subtracted. Shown for TF1, NB4, and HL-60 cells are 10 ng/mL GO cytotoxicity data, whereas 0.25 ng/mL GO data are shown for ML-1 cells, due to their much higher GO sensitivity (see “Results”). GO exposures were without or with 75 μM (TF1) or 50 μM (all other cell lines) PK 11195 (PK), 2.5 μg/mL CSA, or 10 μM MK-571 (MK). PK11195 binding assays were performed in triplicate, and toxicity assays were performed with 2 to 3 replicates for each treatment condition. Additional cytotoxicity data are presented for TF1 and HL-60 cells in Figures 2 and 3. Additional experimental details are presented in “Materials and methods.”
TF1 refers to Pgp+ and MRP+ cells; ML-1, to cells that are MDR−; NB4, to cells that are MRP+/−; and HL-60, to cells that are MRP+. Untrtd indicates untreated; —, not calculated; NS, not significant (ie, P ≥ .05 compared to treatment with GO or sensitizer only).
Compared to viability of untreated cells.
Compared to cells treated with GO only.
Compared to treatment with sensitizer alone.