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Erlotinib inhibits osteolytic bone invasion of human non-small-cell lung cancer cell line NCI-H292.

Furugaki K, Moriya Y, Iwai T, Yorozu K, Yanagisawa M, Kondoh K, Fujimoto-Ohuchi K, Mori K - Clin. Exp. Metastasis (2011)

Bottom Line: In this model, NCI-H292 cells markedly activated osteoclasts in tibia, which resulted in osteolytic bone destruction.Furthermore, erlotinib also inhibited osteoblast/stromal cell proliferation in vitro and the development of osteoclasts induced by RANKL in vitro.In conclusion, erlotinib inhibits tumor-induced osteolytic invasion in bone metastasis by suppressing osteoclast activation through inhibiting tumor growth at the bone metastatic sites, osteolytic factor production in tumor cells, osteoblast/stromal cell proliferation and osteoclast differentiation from mouse bone marrow cells.

View Article: PubMed Central - PubMed

Affiliation: Product Research Department, Kamakura Research Laboratories, Chugai Pharmaceutical Co., Ltd., Kajiwara, Kamakura, Kanagawa, Japan.

ABSTRACT
Previous preclinical and clinical findings have suggested a potential role of epidermal growth factor receptor (EGFR) in osteoclast differentiation and the pathogenesis of bone metastasis in cancer. In this study, we investigated the effect of erlotinib, an orally active EGFR tyrosine kinase inhibitor (TKI), on the bone invasion of human non-small-cell lung cancer (NSCLC) cell line NCI-H292. First, we established a novel osteolytic bone invasion model of NCI-H292 cells which was made by inoculating cancer cells into the tibia of scid mice. In this model, NCI-H292 cells markedly activated osteoclasts in tibia, which resulted in osteolytic bone destruction. Erlotinib treatment suppressed osteoclast activation to the basal level through suppressing receptor activator of NF-κB ligand (RANKL) expression in osteoblast/stromal cell at the bone metastatic sites, which leads to inhibition of osteolytic bone destruction caused by NCI-H292 cells. Erlotinib inhibited the proliferation of NCI-H292 cells in in vitro. Erlotinib suppressed the production of osteolytic factors, such as parathyroid hormone-related protein (PTHrP), IL-8, IL-11 and vascular endothelial growth factor (VEGF) in NCI-H292 cells. Furthermore, erlotinib also inhibited osteoblast/stromal cell proliferation in vitro and the development of osteoclasts induced by RANKL in vitro. In conclusion, erlotinib inhibits tumor-induced osteolytic invasion in bone metastasis by suppressing osteoclast activation through inhibiting tumor growth at the bone metastatic sites, osteolytic factor production in tumor cells, osteoblast/stromal cell proliferation and osteoclast differentiation from mouse bone marrow cells.

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Effects of erlotinib on cell proliferation of NCI-H292 cells. a NCI-H292 tumor cells grew in tibia of mice treated with or without 75 mg/kg of erlotinib. Histological pictures on day 14 stained with HE. Arrowheads indicate the metastatic tumor area. b The metastatic tumor area were measured. Each point represents the mean + SE of sex tuplicates. c The number of Ki-67-positive tumor cells was counted. Each point represents the mean + SE of sex tuplicates. d In vitro cell proliferation determined by MTT assay after 4 days of treatment with erlotinib. Each point represents the mean + SD of triplicates. e Immunoblots of cell lysates from cells treated with or without erlotinib. f Histological pictures on day 14 stained with phosphorylated EGFR
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Fig3: Effects of erlotinib on cell proliferation of NCI-H292 cells. a NCI-H292 tumor cells grew in tibia of mice treated with or without 75 mg/kg of erlotinib. Histological pictures on day 14 stained with HE. Arrowheads indicate the metastatic tumor area. b The metastatic tumor area were measured. Each point represents the mean + SE of sex tuplicates. c The number of Ki-67-positive tumor cells was counted. Each point represents the mean + SE of sex tuplicates. d In vitro cell proliferation determined by MTT assay after 4 days of treatment with erlotinib. Each point represents the mean + SD of triplicates. e Immunoblots of cell lysates from cells treated with or without erlotinib. f Histological pictures on day 14 stained with phosphorylated EGFR

Mentions: Administration of erlotinib tended to inhibit the metastatic tumor growth in this osteolytic bone invasion model (P = 0.24) (Fig. 3a, b). The reduced cell proliferative activity of NCI-H292 cells was observed by immunostaining of proliferative markers Ki-67 (P = 0.24) (Fig. 3c). Furthermore, erlotinib showed a marked inhibition of NCI-H292 cell proliferation in vitro in a dose-dependent manner, and the IC50 value was 0.11 μM (Fig. 3d). An addition of 1 μM of erlotinib markedly blocked not only EGFR phosphorylation but also ERK and Akt phosphorylation which are the downstream of EGFR signaling cascades in NCI-H292 cells (Fig. 3e). Meanwhile, phosphorylated EGFR expression in tumor cells at the metastatic sites was detected in not only the vehicle treatment but also the erlotinib treatment, a difference of the level of EGFR phosphorylation was not observed with or without erlotinib treatment (Fig. 3f).Fig. 3


Erlotinib inhibits osteolytic bone invasion of human non-small-cell lung cancer cell line NCI-H292.

Furugaki K, Moriya Y, Iwai T, Yorozu K, Yanagisawa M, Kondoh K, Fujimoto-Ohuchi K, Mori K - Clin. Exp. Metastasis (2011)

Effects of erlotinib on cell proliferation of NCI-H292 cells. a NCI-H292 tumor cells grew in tibia of mice treated with or without 75 mg/kg of erlotinib. Histological pictures on day 14 stained with HE. Arrowheads indicate the metastatic tumor area. b The metastatic tumor area were measured. Each point represents the mean + SE of sex tuplicates. c The number of Ki-67-positive tumor cells was counted. Each point represents the mean + SE of sex tuplicates. d In vitro cell proliferation determined by MTT assay after 4 days of treatment with erlotinib. Each point represents the mean + SD of triplicates. e Immunoblots of cell lysates from cells treated with or without erlotinib. f Histological pictures on day 14 stained with phosphorylated EGFR
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Fig3: Effects of erlotinib on cell proliferation of NCI-H292 cells. a NCI-H292 tumor cells grew in tibia of mice treated with or without 75 mg/kg of erlotinib. Histological pictures on day 14 stained with HE. Arrowheads indicate the metastatic tumor area. b The metastatic tumor area were measured. Each point represents the mean + SE of sex tuplicates. c The number of Ki-67-positive tumor cells was counted. Each point represents the mean + SE of sex tuplicates. d In vitro cell proliferation determined by MTT assay after 4 days of treatment with erlotinib. Each point represents the mean + SD of triplicates. e Immunoblots of cell lysates from cells treated with or without erlotinib. f Histological pictures on day 14 stained with phosphorylated EGFR
Mentions: Administration of erlotinib tended to inhibit the metastatic tumor growth in this osteolytic bone invasion model (P = 0.24) (Fig. 3a, b). The reduced cell proliferative activity of NCI-H292 cells was observed by immunostaining of proliferative markers Ki-67 (P = 0.24) (Fig. 3c). Furthermore, erlotinib showed a marked inhibition of NCI-H292 cell proliferation in vitro in a dose-dependent manner, and the IC50 value was 0.11 μM (Fig. 3d). An addition of 1 μM of erlotinib markedly blocked not only EGFR phosphorylation but also ERK and Akt phosphorylation which are the downstream of EGFR signaling cascades in NCI-H292 cells (Fig. 3e). Meanwhile, phosphorylated EGFR expression in tumor cells at the metastatic sites was detected in not only the vehicle treatment but also the erlotinib treatment, a difference of the level of EGFR phosphorylation was not observed with or without erlotinib treatment (Fig. 3f).Fig. 3

Bottom Line: In this model, NCI-H292 cells markedly activated osteoclasts in tibia, which resulted in osteolytic bone destruction.Furthermore, erlotinib also inhibited osteoblast/stromal cell proliferation in vitro and the development of osteoclasts induced by RANKL in vitro.In conclusion, erlotinib inhibits tumor-induced osteolytic invasion in bone metastasis by suppressing osteoclast activation through inhibiting tumor growth at the bone metastatic sites, osteolytic factor production in tumor cells, osteoblast/stromal cell proliferation and osteoclast differentiation from mouse bone marrow cells.

View Article: PubMed Central - PubMed

Affiliation: Product Research Department, Kamakura Research Laboratories, Chugai Pharmaceutical Co., Ltd., Kajiwara, Kamakura, Kanagawa, Japan.

ABSTRACT
Previous preclinical and clinical findings have suggested a potential role of epidermal growth factor receptor (EGFR) in osteoclast differentiation and the pathogenesis of bone metastasis in cancer. In this study, we investigated the effect of erlotinib, an orally active EGFR tyrosine kinase inhibitor (TKI), on the bone invasion of human non-small-cell lung cancer (NSCLC) cell line NCI-H292. First, we established a novel osteolytic bone invasion model of NCI-H292 cells which was made by inoculating cancer cells into the tibia of scid mice. In this model, NCI-H292 cells markedly activated osteoclasts in tibia, which resulted in osteolytic bone destruction. Erlotinib treatment suppressed osteoclast activation to the basal level through suppressing receptor activator of NF-κB ligand (RANKL) expression in osteoblast/stromal cell at the bone metastatic sites, which leads to inhibition of osteolytic bone destruction caused by NCI-H292 cells. Erlotinib inhibited the proliferation of NCI-H292 cells in in vitro. Erlotinib suppressed the production of osteolytic factors, such as parathyroid hormone-related protein (PTHrP), IL-8, IL-11 and vascular endothelial growth factor (VEGF) in NCI-H292 cells. Furthermore, erlotinib also inhibited osteoblast/stromal cell proliferation in vitro and the development of osteoclasts induced by RANKL in vitro. In conclusion, erlotinib inhibits tumor-induced osteolytic invasion in bone metastasis by suppressing osteoclast activation through inhibiting tumor growth at the bone metastatic sites, osteolytic factor production in tumor cells, osteoblast/stromal cell proliferation and osteoclast differentiation from mouse bone marrow cells.

Show MeSH
Related in: MedlinePlus