The Regulation Effect Of Monocyte/Macrophagy Chemotactic Factor CCL2 On Osteosarcoma Proliferation,Metastasis And Macrophagy Polarization

Objective: Osteosarcoma is a type of disease seriously harm to human life and health.But the five-years survival rate of osteosarcoma hasn't improved in last 20 years.Lung metastasis is the primary reason to cause death,once it occurred,the five-years survival rate is no more than 30%.Therefore,we need to find therapeutic targets that effectively control the growth and metastasis of osteosarcoma.The expression of CCL2 is related to tumor malignant behaviors in osteosarcoma.So exploration the effects of CCL2 on osteosarcoma proliferation and metastasis,especially the effects and mechanisms in osteosarcoma lung metastasis,are possible to find a therapeutic target in osteosarcoma progress.This study aimed to(1)Exploration the expression of CCL2 in osteosarcoma,analysis the correlation between CCL2 and osteosarcoma lung metastasis;(2)Exploration the effects and mechanisms of CCL2 regulate osteosarcoma proliferation,invasion and migration;(3)Investigation the effects of CCL2 regulates osteosarcoma polarization macrophages to M1 or M2 phenotype.Methods: Detection the differential expression of CCL2 in different lung metastasis potential osteosarcoma cell lines,osteosarcoma animal models and clinical specimens,via RT-PCR,western-blot and immunohistochemistry,research the correlation between the expression of CCL2 and osteosarcoma lung metastasis.Construct stable CCL2 RNA interference osteosarcoma cell lines with lentivirus vectors,perform CCK-8 experiment to research proliferation,wound healing experiment to inverstiage migration,flow cytometry to measure cell cycle and apoptosis,transwell experiment to measure matastases.Co-culture the macrophage with CCL2 RNA interfered osteosarcoma,detecting the mRNA expression of M1 and M2 related gene.Results: CCL2 have detected a highly expression in osteosarcoma cell lines and specimens.Where the expression in lung metastases were higher than primary,and the normal controls were the lowest;high pulmonary metastatic potential osteosarcoma cell lines were higher than low pulmonary metastatic potential osteosarcoma cell lines,and the osteoblast control cells were the lowest.Cell proliferation was slowed down;migration and invasion potential were reduced;Fas L apoptosis were increased.The M1 and M2 related mRNA expression of macrophage cocultured with CCL2 RNA interferition are reduced.Conclusion: This article confirmed the relationship between the expression of CCL2 and osteosarcoma,found CCL2 can promote the formation and metastasis of osteosarcoma,may through resistance the Fas L apoptotic and induce macrophage polarization to promote osteosarcoma growth and survival in the lung microenvironment.CCL2 is a potential therapeutic target to control the growth and lung metastasis of osteosarcoma.