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Insulin-like-growth-factor-binding-protein-3 (IGFBP-3) contrasts melanoma progression in vitro and in vivo.

Naspi A, Panasiti V, Abbate F, Roberti V, Devirgiliis V, Curzio M, Borghi M, Lozupone F, Carotti S, Morini S, Gaudio E, Calvieri S, Londei P - PLoS ONE (2014)

Bottom Line: Insulin-like-factor-binding-protein 3 (IGFBP-3) is known to modulate the activity of insulin-like growth factors (IGFs) besides having a number of IGF-independent effects on cell growth and survival.In the present work, we have evaluated the levels of IGFBP-3 in the blood serum and tissues of patients affected by cutaneous melanoma, showing that loss of IGFBP-3 from both is strongly correlated with disease progression and reduced survival.In summary, IGFBP-3 appears to exert a specific inhibitory effect on melanoma growth and dissemination, suggesting that it may qualify as a useful therapeutic agent in melanomas and perhaps other cancers, at the least as a valid adjuvant therapy during treatment with conventional anti-tumoral drugs.

View Article: PubMed Central - PubMed

Affiliation: Istituto Pasteur-Fondazione Cenci-Bolognetti, Dpt. Biotecnologie Cellulari ed Ematologia, University of Rome Sapienza, Rome, Italy.

ABSTRACT
Insulin-like-factor-binding-protein 3 (IGFBP-3) is known to modulate the activity of insulin-like growth factors (IGFs) besides having a number of IGF-independent effects on cell growth and survival. IGFBP-3 has been reported to decrease significantly in the blood serum of patients affected by certain cancers. In the present work, we have evaluated the levels of IGFBP-3 in the blood serum and tissues of patients affected by cutaneous melanoma, showing that loss of IGFBP-3 from both is strongly correlated with disease progression and reduced survival. In vitro treatment with IGFBP-3 of human and murine metastatic melanoma cell lines specifically inhibited the cells' migratory and invasive behaviour, inducing up-regulation of melanocytic differentiation markers such as tyrosinase activity and melanin content. A molecular analysis of the cellular pathways transducing the effect of IGFBP-3 implicated the Akt-GSK3β axis. Moreover, administration of IGFBP-3 in vivo to SCID mice inoculated with human metastatic melanoma cells strongly reduced or completely inhibited tumor growth. In summary, IGFBP-3 appears to exert a specific inhibitory effect on melanoma growth and dissemination, suggesting that it may qualify as a useful therapeutic agent in melanomas and perhaps other cancers, at the least as a valid adjuvant therapy during treatment with conventional anti-tumoral drugs.

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IGFBP-3 inhibits tumour growth in vivo.(A) Effect of treatment with low (0.37 mg/Kg, green curve) and high (1.87 mg/kg, orange curve) IGFBP-3 doses on the growth over time of Me501 xenografts in SCID mice. Mean results are representative of two different experiments (each experiment on at least 8 mice). Tumor size was measured three times per week with calipers, and volume was calculated as described in the “Methods” section. (B) Immuno-histochemical staining of the cell proliferation marker Ki-67 in Me501 xenografts in tumours excised from SCID mice untreated (A–B) or treated with 1.87 mg/kg IGFBP-3 (C–D). (F) Graph reporting the mean values of Ki-67 positive cells in melanomas from control mice and mice treated with low and high doses of IGFBP-3. The proliferation index was determined as the percentage of proliferating cells (Ki-67 positive cells) in a population of about 800 cells. For this purpose two representative fields were randomly selected in each section and blind-counted by two independent observers. P value was calculated by Mann-Whitney U test.
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pone-0098641-g009: IGFBP-3 inhibits tumour growth in vivo.(A) Effect of treatment with low (0.37 mg/Kg, green curve) and high (1.87 mg/kg, orange curve) IGFBP-3 doses on the growth over time of Me501 xenografts in SCID mice. Mean results are representative of two different experiments (each experiment on at least 8 mice). Tumor size was measured three times per week with calipers, and volume was calculated as described in the “Methods” section. (B) Immuno-histochemical staining of the cell proliferation marker Ki-67 in Me501 xenografts in tumours excised from SCID mice untreated (A–B) or treated with 1.87 mg/kg IGFBP-3 (C–D). (F) Graph reporting the mean values of Ki-67 positive cells in melanomas from control mice and mice treated with low and high doses of IGFBP-3. The proliferation index was determined as the percentage of proliferating cells (Ki-67 positive cells) in a population of about 800 cells. For this purpose two representative fields were randomly selected in each section and blind-counted by two independent observers. P value was calculated by Mann-Whitney U test.

Mentions: To assess the potential clinical relevance of the in vitro results, we evaluated in vivo the anti-tumoral activity of IGFBP-3 using SCID mice xenografted s.c. with Me501 cells. Three days after tumor injection, mice received intraperitoneal (i.p.) injections of saline, or 0.37 mg/Kg or 1.87 mg/kg IGFBP-3 three times a week (TTW) for three weeks and tumor growth was estimated during the follow-up. As illustrated in Fig. 9, the growth of Me501 tumors in mice treated with IGFBP-3 was significantly reduced as compared with control animals, with a dose-dependent efficacy, and the size of IGFBP-3-treated tumors was significantly smaller than control tumors at follow-up (P = 0.0005), indicating that IGFBP-3 treatment may exert an antineoplastic activity in vivo. More in detail, Tumor Volume Inhibition percent (TVI%) in IGFBP-3-treated mice was respectively of 40,33 and 94,61% at 28 days post-tumor implant. At sacrifice, tumor diameter was significantly lower (t-Test p<0.001) in IGFBP-3 (3±1,5 and 4.5±2 mm) in comparison to saline (9.0±3 mm) treated mice. Moreover, complete response (CR) was obtained in 2/8 mice treated with the lower IGFBP-3 concentration, in 3/8 mice treated with the higher IGFBP-3 concentration but in none of the saline group. Four weeks after tumor implant mice were sacrificed and tumors were collected for histological examination. We observed that the tumor mass of treated mice was occupied by large necrotic areas that accounted for most of the tumor size, indicating that the cytotoxic effects of IGFBP-3 were more profound than those anticipated by the simple evaluation of in vivo tumor size (not shown). We also noticed a marked reduction in Ki67 positive cells in the IGFBP-3-treated groups (Fig. 9). Moreover, tumors from IGFBP-3 treated mice were consistently darker than those from untreated mice, suggesting up regulation of melanin synthesis. Indeed, tyrosinase activity measured on lysates of tumour tissues was higher in the tumours excised from IGFBP-3-treated mice, confirming the observations made on cultured melanoma cells (not shown).


Insulin-like-growth-factor-binding-protein-3 (IGFBP-3) contrasts melanoma progression in vitro and in vivo.

Naspi A, Panasiti V, Abbate F, Roberti V, Devirgiliis V, Curzio M, Borghi M, Lozupone F, Carotti S, Morini S, Gaudio E, Calvieri S, Londei P - PLoS ONE (2014)

IGFBP-3 inhibits tumour growth in vivo.(A) Effect of treatment with low (0.37 mg/Kg, green curve) and high (1.87 mg/kg, orange curve) IGFBP-3 doses on the growth over time of Me501 xenografts in SCID mice. Mean results are representative of two different experiments (each experiment on at least 8 mice). Tumor size was measured three times per week with calipers, and volume was calculated as described in the “Methods” section. (B) Immuno-histochemical staining of the cell proliferation marker Ki-67 in Me501 xenografts in tumours excised from SCID mice untreated (A–B) or treated with 1.87 mg/kg IGFBP-3 (C–D). (F) Graph reporting the mean values of Ki-67 positive cells in melanomas from control mice and mice treated with low and high doses of IGFBP-3. The proliferation index was determined as the percentage of proliferating cells (Ki-67 positive cells) in a population of about 800 cells. For this purpose two representative fields were randomly selected in each section and blind-counted by two independent observers. P value was calculated by Mann-Whitney U test.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4048209&req=5

pone-0098641-g009: IGFBP-3 inhibits tumour growth in vivo.(A) Effect of treatment with low (0.37 mg/Kg, green curve) and high (1.87 mg/kg, orange curve) IGFBP-3 doses on the growth over time of Me501 xenografts in SCID mice. Mean results are representative of two different experiments (each experiment on at least 8 mice). Tumor size was measured three times per week with calipers, and volume was calculated as described in the “Methods” section. (B) Immuno-histochemical staining of the cell proliferation marker Ki-67 in Me501 xenografts in tumours excised from SCID mice untreated (A–B) or treated with 1.87 mg/kg IGFBP-3 (C–D). (F) Graph reporting the mean values of Ki-67 positive cells in melanomas from control mice and mice treated with low and high doses of IGFBP-3. The proliferation index was determined as the percentage of proliferating cells (Ki-67 positive cells) in a population of about 800 cells. For this purpose two representative fields were randomly selected in each section and blind-counted by two independent observers. P value was calculated by Mann-Whitney U test.
Mentions: To assess the potential clinical relevance of the in vitro results, we evaluated in vivo the anti-tumoral activity of IGFBP-3 using SCID mice xenografted s.c. with Me501 cells. Three days after tumor injection, mice received intraperitoneal (i.p.) injections of saline, or 0.37 mg/Kg or 1.87 mg/kg IGFBP-3 three times a week (TTW) for three weeks and tumor growth was estimated during the follow-up. As illustrated in Fig. 9, the growth of Me501 tumors in mice treated with IGFBP-3 was significantly reduced as compared with control animals, with a dose-dependent efficacy, and the size of IGFBP-3-treated tumors was significantly smaller than control tumors at follow-up (P = 0.0005), indicating that IGFBP-3 treatment may exert an antineoplastic activity in vivo. More in detail, Tumor Volume Inhibition percent (TVI%) in IGFBP-3-treated mice was respectively of 40,33 and 94,61% at 28 days post-tumor implant. At sacrifice, tumor diameter was significantly lower (t-Test p<0.001) in IGFBP-3 (3±1,5 and 4.5±2 mm) in comparison to saline (9.0±3 mm) treated mice. Moreover, complete response (CR) was obtained in 2/8 mice treated with the lower IGFBP-3 concentration, in 3/8 mice treated with the higher IGFBP-3 concentration but in none of the saline group. Four weeks after tumor implant mice were sacrificed and tumors were collected for histological examination. We observed that the tumor mass of treated mice was occupied by large necrotic areas that accounted for most of the tumor size, indicating that the cytotoxic effects of IGFBP-3 were more profound than those anticipated by the simple evaluation of in vivo tumor size (not shown). We also noticed a marked reduction in Ki67 positive cells in the IGFBP-3-treated groups (Fig. 9). Moreover, tumors from IGFBP-3 treated mice were consistently darker than those from untreated mice, suggesting up regulation of melanin synthesis. Indeed, tyrosinase activity measured on lysates of tumour tissues was higher in the tumours excised from IGFBP-3-treated mice, confirming the observations made on cultured melanoma cells (not shown).

Bottom Line: Insulin-like-factor-binding-protein 3 (IGFBP-3) is known to modulate the activity of insulin-like growth factors (IGFs) besides having a number of IGF-independent effects on cell growth and survival.In the present work, we have evaluated the levels of IGFBP-3 in the blood serum and tissues of patients affected by cutaneous melanoma, showing that loss of IGFBP-3 from both is strongly correlated with disease progression and reduced survival.In summary, IGFBP-3 appears to exert a specific inhibitory effect on melanoma growth and dissemination, suggesting that it may qualify as a useful therapeutic agent in melanomas and perhaps other cancers, at the least as a valid adjuvant therapy during treatment with conventional anti-tumoral drugs.

View Article: PubMed Central - PubMed

Affiliation: Istituto Pasteur-Fondazione Cenci-Bolognetti, Dpt. Biotecnologie Cellulari ed Ematologia, University of Rome Sapienza, Rome, Italy.

ABSTRACT
Insulin-like-factor-binding-protein 3 (IGFBP-3) is known to modulate the activity of insulin-like growth factors (IGFs) besides having a number of IGF-independent effects on cell growth and survival. IGFBP-3 has been reported to decrease significantly in the blood serum of patients affected by certain cancers. In the present work, we have evaluated the levels of IGFBP-3 in the blood serum and tissues of patients affected by cutaneous melanoma, showing that loss of IGFBP-3 from both is strongly correlated with disease progression and reduced survival. In vitro treatment with IGFBP-3 of human and murine metastatic melanoma cell lines specifically inhibited the cells' migratory and invasive behaviour, inducing up-regulation of melanocytic differentiation markers such as tyrosinase activity and melanin content. A molecular analysis of the cellular pathways transducing the effect of IGFBP-3 implicated the Akt-GSK3β axis. Moreover, administration of IGFBP-3 in vivo to SCID mice inoculated with human metastatic melanoma cells strongly reduced or completely inhibited tumor growth. In summary, IGFBP-3 appears to exert a specific inhibitory effect on melanoma growth and dissemination, suggesting that it may qualify as a useful therapeutic agent in melanomas and perhaps other cancers, at the least as a valid adjuvant therapy during treatment with conventional anti-tumoral drugs.

Show MeSH
Related in: MedlinePlus