Fig. 2: Both IBSP and miR-19a are essential for bone metastasis of ER+ breast cancer. | Nature Communications

Fig. 2: Both IBSP and miR-19a are essential for bone metastasis of ER+ breast cancer.

From: Exosomal miR-19a and IBSP cooperate to induce osteolytic bone metastasis of estrogen receptor-positive breast cancer

Fig. 2

a miR-19a and/or IBSP were ectopically expressed in MCF7, and they were transplanted into mammary fat pads of female nude mice. The growth of the tumor was monitored by measuring the luciferase activity by IVIS Bioimager. p = 0.3110 (MCF7/pLenti/GFP vs MCF7/IBSP/miR-19a; n = 6 vs 6), p = 0.2649 (MCF7/pLenti/GFP vs MCF7/IBSP/GFP; n = 6 vs 6), p = 0.4739 (MCF7/pLenti/GFP vs MCF7/pLenti/miR-19a; n = 6 vs 8). Data are presented as mean values ± SEM. b At day 21, tumors were removed from the mammary fat pad and their weights were measured. p = 0.3971 (MCF7/pLenti/GFP vs MCF7/IBSP/miR-19a; n = 6 vs 6), p = 0.2178 (MCF7/pLenti/GFP vs MCF7/IBSP/GFP; n = 6 vs 6), p = 0.7632 (MCF7/pLenti/GFP vs MCF7/pLenti/miR-19a; n = 6 vs 8). Data are presented as mean values ± SEM. c miR-19a and/or IBSP were ectopically expressed, and they were transplanted into female nude mice via intracardiac injection. The incidence of bone metastasis was monitored by measuring the luciferase activity by IVIS Bioimager. Log-rank (Mantel–Cox) test was performed to calculate the p-value. p = 0.0105 (MCF7/pLenti/GFP vs MCF7/IBSP/miR-19a), p = 0.3982 (MCF7/pLenti/GFP vs MCF7/IBSP/GFP), p = 0.9230 (MCF7/pLenti/GFP vs MCF7/pLenti/miR-19a). d The legs of mice were imaged by X-ray and the bone density was measured by ImageJ. p = 0.0007 (MCF7/pLenti/GFP vs MCF7/IBSP/miR-19a; n = 6 vs 12), p = 0.8621 (MCF7/pLenti/GFP vs MCF7/IBSP/GFP, n = 6 vs 7), p = 0.6397 (MCF7/pLenti/GFP vs MCF7/pLenti/miR-19a, n = 6 vs 7). Two-sided student’s t-tests were performed. Data are presented as mean values ± SEM. e TRAP staining was performed in tumor-bearing bones from mice. The total OC surface area relative to the bone surface area was measured by ImageJ, then calculated and compared among different groups. H&E staining of the same field was shown together with the TRAP staining. Scale bar, 100 µm. p < 0.0001 (MCF7/pLenti/GFP vs MCF7/IBSP/miR-19a, n = 6 vs 12), p = 0.1598 (MCF7/pLenti/GFP vs MCF7/IBSP/GFP, n = 6 vs 7), p = 0.0867(MCF7/pLenti/GFP vs MCF7/pLenti/miR-19a, n = 6 vs 7). Two-sided student’s t-tests were performed. Data are presented as mean values ± SEM. f miR-19a and/or IBSP were ectopically expressed, and they were transplanted into female nude mice via intra-tibia injection. The growth of tumor in bone was monitored by measuring the luciferase activity by IVIS Bioimager. p = 0.0175 (T47D/pLenti/GFP vs T47D/IBSP/miR-19a), p = 0.4877 (T47D/pLenti/GFP vs T47D/IBSP/GFP), p = 0.3305 (T47D/pLenti/GFP vs T47D/pLenti/miR-19a). Data are presented as mean values ± SEM. g The legs of mice were imaged by X-ray and the bone density was measured by ImageJ. p = 0.0034 (T47D/pLenti/GFP vs T47D/IBSP/miR-19a n = 5 vs 6), p = 0.5750 (T47D/pLenti/GFP vs T47D/IBSP/GFP, n = 5 vs 4), p = 0.3809 (T47D/pLenti/GFP vs T47D/pLenti/miR-19a, n = 5 vs 6). Two-sided student’s t-tests were performed. Data are presented as mean values ± SEM. h TRAP staining was performed in tumor-bearing bones from the mice. The OC surface relative to the bone surface was calculated and compared among different groups. H&E staining of the same field was shown together with the TRAP staining. Scale bar, 100 µm. p = 0.0001 (T47D/pLenti/GFP vs T47D/IBSP/miR-19a, n = 5 vs 6), p = 0.7711 (T47D/pLenti/GFP vs T47D/IBSP/GFP, n = 5 vs 4), p = 0.5701 (T47D/pLenti/GFP vs T47D/pLenti/miR-19a, n = 5 vs 6). Two-sided student’s t-tests were performed. Data are presented as mean values ± SEM.

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