American Society of Hematology

Figure 6

$Figure 6. The inducible expression of S1PR2 delays tumor growth in vivo. (A-F) NOD/SCID/IL2Rγ−/− mice were subcutaneously inoculated in both flanks with 1 × 107 SU-DHL6 cells that had been transduced with pIND21-S1PR2 and sorted for enhanced GFP (eGFP). eGFP-negative cells were used as a negative control. Mice were switched to doxycycline-containing chow once tumors were palpable (on day 13 posttransplantation). Mice transplanted with eGFP-positive cells were maintained on normal chow as another negative control. The data shown were pooled from 2 independent experiments. (A) S1PR2 expression as determined by qRT-PCR (normalized to ACTIN) of resected tumors. (B-C) Tumor volume as determined over time (days posttransplantation, B; P values were calculated using 2-way analysis of variance with Tukey multiple comparison test) and at the study end point (C). (D) Tumor weight at the study end point. (E) Representative excised tumors of the indicated treatment groups. (F) eGFP-positive fraction of tumor cells at the study end point. (G-M) 1 × 107 subcutaneously passaged SU-DHL6 (pIND21-S1PR2) cells were sorted for eGFP expression and injected into the tail veins of NOD/SCID/IL2Rγ−/− mice; mice were treated with doxycycline starting on day 15 post injection as described in panel A-F. (G) Body weight per mouse as recorded every 3 days for the 3 treatment arms (means ± SEM; P values were calculated using 2-way ANOVA with Tukey multiple comparison test). (H) Body weight change relative to day 15 post–tumor cell injection. (I-J) Spleen weight and tumor cell burden per milligram of spleen at the study end point. (K) Tumor cell burden in the blood at the study end point, as determined by CD45 staining. (L-M) Fraction of eGFP-positive cells in % of all human CD45-positive tumor cells in the spleens (L) and blood (M) of the indicated groups at the study end point. Horizontal lines indicate the medians, each symbol represents one tumor. *P < .05, **P < .01, and ***P < .001 (2-tailed Mann-Whitney U test for all panels except B, G, and N). (N) Kaplan-Meyer plot of S1PR2+/+/Emu-MYC-transgenic (MYC-tg), S1PR2+/− and S1PR2+/−/MYC-tg mice (4 per group). **P < .01; calculated with log-rank (Mantel-Cox) test. DOX, doxycycline; n.s., not significant.$

The inducible expression of S1PR2 delays tumor growth in vivo. (A-F) NOD/SCID/IL2Rγ^−/− mice were subcutaneously inoculated in both flanks with 1 × 10⁷ SU-DHL6 cells that had been transduced with pIND21-S1PR2 and sorted for enhanced GFP (eGFP). eGFP-negative cells were used as a negative control. Mice were switched to doxycycline-containing chow once tumors were palpable (on day 13 posttransplantation). Mice transplanted with eGFP-positive cells were maintained on normal chow as another negative control. The data shown were pooled from 2 independent experiments. (A) S1PR2 expression as determined by qRT-PCR (normalized to ACTIN) of resected tumors. (B-C) Tumor volume as determined over time (days posttransplantation, B; P values were calculated using 2-way analysis of variance with Tukey multiple comparison test) and at the study end point (C). (D) Tumor weight at the study end point. (E) Representative excised tumors of the indicated treatment groups. (F) eGFP-positive fraction of tumor cells at the study end point. (G-M) 1 × 10⁷ subcutaneously passaged SU-DHL6 (pIND21-S1PR2) cells were sorted for eGFP expression and injected into the tail veins of NOD/SCID/IL2Rγ^−/− mice; mice were treated with doxycycline starting on day 15 post injection as described in panel A-F. (G) Body weight per mouse as recorded every 3 days for the 3 treatment arms (means ± SEM; P values were calculated using 2-way ANOVA with Tukey multiple comparison test). (H) Body weight change relative to day 15 post–tumor cell injection. (I-J) Spleen weight and tumor cell burden per milligram of spleen at the study end point. (K) Tumor cell burden in the blood at the study end point, as determined by CD45 staining. (L-M) Fraction of eGFP-positive cells in % of all human CD45-positive tumor cells in the spleens (L) and blood (M) of the indicated groups at the study end point. Horizontal lines indicate the medians, each symbol represents one tumor. *P < .05, **P < .01, and ***P < .001 (2-tailed Mann-Whitney U test for all panels except B, G, and N). (N) Kaplan-Meyer plot of S1PR2^+/+/Emu-MYC-transgenic (MYC-tg), S1PR2^+/− and S1PR2^+/−/MYC-tg mice (4 per group). **P < .01; calculated with log-rank (Mantel-Cox) test. DOX, doxycycline; n.s., not significant.

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