Establishment and characterization of a murine osteosarcoma cell line (LM8) with high metastatic potential to the lung

Asai, Tatsuya; Ueda, Takafumi; Itoh, Kazuyuki; Yoshioka, Kiyoko; Aoki, Yasuaki; Mori, Shigeki; Yoshikawa, Hideki

doi:10.1002/(sici)1097-0215(19980504)76:3<418::aid-ijc21>3.0.co;2-5

Cited by 184 publications

(200 citation statements)

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“…The LM8 cell line was derived from the original Dunn cell line 24 by in vivo selection. 22 In this study, LM8 cells were seeded at a density of 2.0 Â 10 6 cells in 175 cm 2 culture flasks. When 490% confluency was reached at day 2, the cells were digested using 0.05% trypsin and encapsulated in 1.2% low-viscosity alginate (Keltone LV; Kelco, Chicago, IL) in 0.15 M sodium chloride (NaCl) at a concentration of 4.0 Â 10 6 cells per ml.…”

Section: Methodsmentioning

confidence: 99%

“…26 The LM8 parent Dunn cell line 22 was also evaluated. The NF-kB (p65) signaling activity of untreated LM8 cells (control) was significantly higher (Po0.01), about threefold, than that of the Dunn cells (Figure 2).…”

Section: Nf-kb Dna-binding Capacitymentioning

confidence: 99%

“…21 Therefore, we hypothesized that transfection with NF-B decoy ODN would suppress the tumor growth and pulmonary metastases of osteosarcoma. Using murine osteosarcoma cells from two metastatic clones, the parental Dunn cell line and its derivative LM8 with greater metastatic potential to the lung, 22 we have recently established a novel alginate-encapsulated tumor spheroid model 23 to mimic the in vivo microenvironment. The purpose of this study was to examine the effects of NF-B decoy ODN on tumor progression and metastasis-related gene expression in vitro, as well as pulmonary metastasis in vivo, using the LM8 cell line in the alginateencapsulated tumor spheroid model.…”

Section: Introductionmentioning

confidence: 99%

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Transfection of NF-κB decoy oligodeoxynucleotide suppresses pulmonary metastasis by murine osteosarcoma

et al. 2010

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Nuclear factor-kappa B (NF-kB) has a pivotal role in the progression and distant metastasis of cancers, including malignant bone tumors. To inhibit NF-kB activation, a new molecular therapy using synthetic double-stranded oligodeoxynucleotide (ODN) as a 'decoy' cis element against NF-kB has been developed. To determine whether pulmonary metastasis of osteosarcoma is reduced by inhibiting the action of NF-kB, NF-kB decoy ODN was transfected into the nuclei of murine osteosarcoma cells with high pulmonary metastatic potential, the LM8 cell line, using a three-dimensional alginate spheroid culture model. An in vitro study demonstrated the successful transfection of LM8 cells cultured in alginate beads by 'naked' NF-kB decoy ODN and that the activation of NF-kB signaling was significantly suppressed. Tumor growth was not affected by transfection of NF-kB decoy ODN, however, the expression of vascular endothelial growth factor (VEGF) and intercellular adhesion molecule 1 (ICAM-1) mRNA was markedly decreased. Furthermore, the transfection of 'naked' NF-kB decoy ODN effectively suppressed pulmonary metastasis in an in vivo alginate bead transplantation model. Our results suggest that NF-kB has a central and specific role in the regulation of tumor metastasis and could be a molecular target for development of anti-metastatic treatments for osteosarcoma.

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Section: Methodsmentioning

confidence: 99%

Section: Nf-kb Dna-binding Capacitymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Transfection of NF-κB decoy oligodeoxynucleotide suppresses pulmonary metastasis by murine osteosarcoma

et al. 2010

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“…P-glycoprotein (P-gp)-overexpressing cell line which was established by stepwise increments of DOX, MOS/ADR was generated as previously reported (Takeshita et al, 1996). Murine osteosarcoma cell line, LM8 was established from the murine Dunn osteosarcoma cell line (Asai et al, 1998). Normal human dermal fibroblasts (NHDF) cell line was purchased from Kurabo (Osaka, Japan).…”

Section: Cell Linesmentioning

confidence: 99%

Combined effects of a third-generation bisphosphonate, zoledronic acid with other anticancer agents against murine osteosarcoma

et al. 2007

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Bisphosphonates (BPs) are widely used to treat bone diseases and also appear to possess direct antitumour activity. We have previously reported that third-generation BPs such as zoledronic acid (ZOL) and minodronic acid (YM529) synergistically augment the effects of anticancer agents in various cancer cells. Recently, we have also reported the antitumour effects of YM529 on murine osteosarcoma cells. As YM529 has not been clinically available, we herein focused on the anti-osteosarcoma effects of ZOL which is clinically available. In addition to ZOL alone, we evaluated the concurrent or sequential combined effects of ZOL with other anticancer agents against murine osteosarcoma cell lines. ZOL showed almost same anti-osteosarcoma activity compared with YM529 and more sensitive growth inhibitory effects against osteosarcoma cells than normal cells. Moreover, ZOL acted synergistically in vitro when administered concurrently with paclitaxel (PAC) or gemcitabine (GEM), not only in wild-type osteosarcoma cells but also in P-glycoprotein (P-gp)-overexpressing osteosarcoma cells, which were much less sensitive against each anticancer agent. Furthermore, 24 h of ZOL pretreatment significantly augmented the sensitivity of doxorubicin (DOX), PAC or GEM against osteosarcoma cells. These findings suggest that combined administration of ZOL with other anticancer agents may improve the osteosarcoma treatment.

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“…Spontaneous microscopic lung metastases become evident 4 weeks after subcutaneous implantation of 5 3 10 6 LM8 cells into syngeneic immunocompetent C3H mice. 10 Both cell lines were maintained in DMEM media supplemented with nonessential amino acids, sodium pyruvate, L-glutamine and 10% fetal bovine serum. Cells were verified to be negative for mycoplasma using the Mycoplasma Plus PCR Primer set (Stratagen, Inc., La Jolla, CA) and for pathogenic murine viruses (M.A.…”

Section: Cell Lines and Animal Modelsmentioning

confidence: 99%

Aerosol gemcitabine inhibits the growth of primary osteosarcoma and osteosarcoma lung metastases

Koshkina

Kleinerman

2005

Intl Journal of Cancer

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Osteosacarcoma (OS) lung metastases are often resistant to chemotherapy. Most anticancer drugs are administered systemically. In many cases this is followed by dose-dependent toxicity, which may not allow the achievement of therapeutic levels in lungs to eradicate metastases. We determined the efficacy of gemcitabine (GCB) by administering it directly to the lungs via aerosol and studied the role of the Fas pathway in response to the therapy. We used 2 osteosarcoma lung metastases animal models: human LM7 cells that form lung metastases in mice following intravenous injection and murine LM8 cells, which grows subcutaneously in mice and spontaneously metastasize to the lung. Treatment was initiated when the presence of lung metastases had been established. Aerosol GCB inhibited the growth of lung metastases in mice. Intraperitoneal GCB administration at similar dosage had no effect on lung metastases. Besides its direct effect on lung metastases, aerosol GCB suppressed the growth of subcutaneous LM8 tumor. Histopathological examination of mice receiving aerosol GCB showed no evidence of toxicity. Lungs are distinguished from other tissues by the constitutive expression of FasL. Since exposure of tumor cells to GCB upregulated Fas expression, we hypothesized that the susceptibility of the tumor cells to ligandinduced cell death by resident lung cells may be increased. Therefore, the Fas pathway may contribute to the therapeutic effect of aerosol GCB. ' 2005 Wiley-Liss, Inc. Key words: Fas; lung environment; toxicityThe most common site for metastatic spread of OS is the lungs. Patients with OS lung metastases have a poor prognosis with limited therapeutic options. Metastatic and relapsed disease is often resistant to salvage chemotherapy. Our laboratory's goal is to identify new therapeutic approaches for these patients.GCB has been shown to have limited efficacy in the treatment of advanced sarcomas, including OS, 1 but this may be explained in part because systemic administration of GCB does not achieve therapeutic levels in the lungs. We hypothesized that targeted delivery of GCB to the lungs via aerosol will circumvent this problem, yielding a higher drug concentration in the area of the tumor and offer a unique therapeutic opportunity for patients with lung metastases. Indeed, we previously demonstrated that the intranasal administration of GCB at a dose 8-fold lower than the i.v. dose induced the regression of established OS lung metastases. 2 However, the intranasal delivery of drugs to the lungs in humans is not ideal as the drugs will not reach the peripheral regions of the lungs where metastases often develop.Aerosol delivery can bypass this limitation. The clinical feasibility of aerosol drug delivery has been demonstrated with both 9-nitrocamptothecin and GM-CSF. 3,4 In our study, we investigated the efficacy of aerosol GCB in treating OS lung metastases using 2 OS animal model. Aerosol GCB significantly inhibited the growth of primary tumors and of established lung metastases and also prevented metast...

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Establishment and characterization of a murine osteosarcoma cell line (LM8) with high metastatic potential to the lung

Cited by 184 publications

References 23 publications

Transfection of NF-κB decoy oligodeoxynucleotide suppresses pulmonary metastasis by murine osteosarcoma

Transfection of NF-κB decoy oligodeoxynucleotide suppresses pulmonary metastasis by murine osteosarcoma

Combined effects of a third-generation bisphosphonate, zoledronic acid with other anticancer agents against murine osteosarcoma

Aerosol gemcitabine inhibits the growth of primary osteosarcoma and osteosarcoma lung metastases

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