![Figure 6. miRNA-15a and -16 target MM cells in the context of bone marrow (BM) milieu in vitro and in vivo. MM cells (pre-miRNA-15a-, -16-1 precursors probe–, control probe–transfected MM.1S cells) were harvested at 24 hours after transfection. Nontransfected MM.1S cells were used as control (ctrl). (A) Transwell migration assay. SDF-1 (30 nM) was placed in the lower chambers and migration was determined after 2 hours. Control probe–transfected and not-transfected cells showed significant migration with SDF-1 30 nM, whereas the pre-miRNA-15a and -16-1 probe–transfected cells showed minimal migration in response to SDF-1. (B) Adhesion assay to primary BMSCs. Control probe–transfected and nontransfected cells showed significant increase in adhesion to BMSCs, compared with control (BMSCs-non coated wells). Pre-miR-15a and -16-1 probe–transfected cells showed lower adhesion to BMSCs. (C) MM cells (pre-miRNA-15a–, -16-1 probe–, control probe–transfected MM.1S cells) cells were cultured for 48 hours in the presence or absence of BMSCs, and cell proliferation was assessed using [3H]-thymidine uptake assay. All data represent mean (SD) of triplicate experiments. (D) In vivo confocal imaging. GFP+ MM cells were transfected using either control-probe or pre-miRNA-15a, -16-1 probes, and then injected in mice 24 hours after transfection. GFP+ MM cells (green color) were excited with a 491-nm solid-state laser. Blood vessels (red color) were imaged using Evans blue excited with a 635-nm laser. Images of parasagittal vasculature and GFP+ tumor cells in the mouse skull BM. Numbers 1 through 8 indicate the 8 areas of the BM niches that were imaged. Control probe–transfected cells homed and adhered more than the miRNA-15a, -16-1–transfected counterparts. (E) Bioluminescence imaging (BLI) demonstrates tumor progression in the control mice with minimal to no BLI signal in the mice injected with miRNA-15a, -16-1–transfected MM cells. (a: dorsal image; b: ventral image). BLI signal has been quantitated using ImageJ software (P values are shown; *, **P < .001).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/113/26/10.1182_blood-2009-01-198408/4/zh89990938240006.jpeg?Expires=1722409900&Signature=37ZVMvlWtcA2iqjEyKgg-cHQHgktBrCV6zZntdlcrOSBXVsHNyZbSsU0ZH8iGF~0PyV5N3QQsN3W6xDsBGw-hHitAuyinVm4rdiRI74T3GQmMsZRkX9-iJ38HU6ktyYAtJ7uVqlIgCJWv-LqE7gEiENERiQfreyIco9-BHXgqcRj-WMEM3oJJlQFIn3p-mZcnunTeA~0nNnWgY-gpNucS4PENVxKkm2JBH1hbrBWAxUXP5jx3MufpAFcl4a1KKKmloofOW9NLB5BH9gxR3ix21iXYv10I9nA97vws-MjgrAVywSPZGEx3KS1j90-gNq1Davv9RFRFngxBhDy09bpsQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
miRNA-15a and -16 target MM cells in the context of bone marrow (BM) milieu in vitro and in vivo. MM cells (pre-miRNA-15a-, -16-1 precursors probe–, control probe–transfected MM.1S cells) were harvested at 24 hours after transfection. Nontransfected MM.1S cells were used as control (ctrl). (A) Transwell migration assay. SDF-1 (30 nM) was placed in the lower chambers and migration was determined after 2 hours. Control probe–transfected and not-transfected cells showed significant migration with SDF-1 30 nM, whereas the pre-miRNA-15a and -16-1 probe–transfected cells showed minimal migration in response to SDF-1. (B) Adhesion assay to primary BMSCs. Control probe–transfected and nontransfected cells showed significant increase in adhesion to BMSCs, compared with control (BMSCs-non coated wells). Pre-miR-15a and -16-1 probe–transfected cells showed lower adhesion to BMSCs. (C) MM cells (pre-miRNA-15a–, -16-1 probe–, control probe–transfected MM.1S cells) cells were cultured for 48 hours in the presence or absence of BMSCs, and cell proliferation was assessed using [3H]-thymidine uptake assay. All data represent mean (SD) of triplicate experiments. (D) In vivo confocal imaging. GFP+ MM cells were transfected using either control-probe or pre-miRNA-15a, -16-1 probes, and then injected in mice 24 hours after transfection. GFP+ MM cells (green color) were excited with a 491-nm solid-state laser. Blood vessels (red color) were imaged using Evans blue excited with a 635-nm laser. Images of parasagittal vasculature and GFP+ tumor cells in the mouse skull BM. Numbers 1 through 8 indicate the 8 areas of the BM niches that were imaged. Control probe–transfected cells homed and adhered more than the miRNA-15a, -16-1–transfected counterparts. (E) Bioluminescence imaging (BLI) demonstrates tumor progression in the control mice with minimal to no BLI signal in the mice injected with miRNA-15a, -16-1–transfected MM cells. (a: dorsal image; b: ventral image). BLI signal has been quantitated using ImageJ software (P values are shown; *, **P < .001).
