Lung cells support osteosarcoma cell migration and survival

Abstract

BackgroundOsteosarcoma (OS) is the most common primary bone tumor, with a propensity to metastasize to the lungs. Five-year survival for metastatic OS is below 30%, and has not improved for several decades despite the introduction of multi-agent chemotherapy. Understanding OS cell migration to the lungs requires an evaluation of the lung microenvironment. Here we utilized an in vitro lung cell and OS cell co-culture model to explore the interactions between OS and lung cells, hypothesizing that lung cells would promote OS cell migration and survival. The impact of a novel anti-OS chemotherapy on OS migration and survival in the lung microenvironment was also examined.MethodsThree human OS cell lines (SJSA-1, Saos-2, U-2) and two human lung cell lines (HULEC-5a, MRC-5) were cultured according to American Type Culture Collection recommendations. Human lung cell lines were cultured in growth medium for 72 h to create conditioned media. OS proliferation was evaluated in lung co-culture and conditioned media microenvironment, with a murine fibroblast cell line (NIH-3 T3) in fresh growth medium as controls. Migration and invasion were measured using a real-time cell analysis system. Real-time PCR was utilized to probe for Aldehyde Dehydrogenase (ALDH1) expression. Osteosarcoma cells were also transduced with a lentivirus encoding for GFP to permit morphologic analysis with fluorescence microscopy. The anti-OS efficacy of Disulfiram, an ALDH-inhibitor previously shown to inhibit OS cell proliferation and metastasis in vitro, was evaluated in each microenvironment.ResultsLung-cell conditioned medium promoted osteosarcoma cell migration, with a significantly higher attractive effect on all three osteosarcoma cell lines compared to basic growth medium, 10% serum containing medium, and NIH-3 T3 conditioned medium (p <0.05). Lung cell conditioned medium induced cell morphologic changes, as demonstrated with GFP-labeled cells. OS cells cultured in lung cell conditioned medium had increased alkaline phosphatase staining.ConclusionsLung endothelial HULEC-5a cells are attractants for OS cell migration, proliferation, and survival. The SJSA-1 osteosarcoma cell line demonstrated greater metastatic potential than Saos-2 and U-2 cells. ALDH appears to be involved in the interaction between lung and OS cells, and ALP may be a valuable biomarker for monitoring functional OS changes during metastasis.

<p>circPVT1 Facilitates Invasion and Metastasis by Regulating miR-205-5p/c-FLIP Axis in Osteosarcoma</p>

“…4 A considerable proportion of patients already exhibit metastasis at the time of diagnosis, and the most frequent target organ of metastasis is lung. 5,6 The subsequent respiratory impairment is a leading cause of the death of OS patients. 7 These lung-metastasis OS patients have the poorest overall prognosis, with only a 19% five-year survival rate.…”

ONZIN Upregulation by Mutant p53 Contributes to Osteosarcoma Metastasis Through the CXCL5-MAPK Signaling Pathway

“…Osteosarcoma is the most common primary bone tumor in young adults [1,2] and frequently occurs in the femur (42%), tibia (19%), and humerus (10%) [3]. Since the 1970s, despite the use of multi-agent chemotherapy and improvement in surgical techniques, the 5-year overall survival rate of osteosarcoma remains approximately 60% [4].…”

Untangling the response of bone tumor cells and bone forming cells to matrix stiffness and adhesion ligand density by means of hydrogels

“…However, in the hydrogels with the same stiffness, positive staining and cell numbers varied little with the change in the density of adhesion ligands. ALDH plays a vital role in clinical osteosarcoma metastasis[24], and CD133 is also an osteosarcoma marker of metastasis and invasion[33, 34]. As shown in Fig.…”

“…The activity of ALDH that is a marker for the malignancy of osteosarcoma [24] was tested. Briefly, after intrascaffold cultured for 7 days, the cell-gel composites of osteosarcoma were first placed on ice then ALDH was detected by an ALDH test kit (Solarbio, Beijing, China) as indicated by the manufacturer.…”