Study On CCL18 Regulates Osteosarcoma Metastasis And Breast Cancer Bone Metastasis

OBJECTIVE:1.To investigate the biological role and related molecular mechanism of Macrophage-derived CCL18 in the initiation and progression ofosteosarcoma(OS).2.To investigate the expression of macrophage-derived chemokine CCL18 in bone metastasis tissues of breast cancer and its correlation with the median time of bone metastases in patients3.To investigate the clinical efficacy and safety of percutaneous vertebroplasty(PVP)in the treatment of breast cancer patients with spinal metastases.METHODS:1.Immunohistochemical double staining was used to detect the expression of CCL18 + tumor-associated macrophages(TAMs)in both lung metastases and primary tissues of osteosarcoma.ELISA was used to detect the expression of CCL18 in sera of osteosarcoma patients and healthy people.2.CCK8,wound-healing assay,Transwell chamber assay were performed to detect the effect of CCL18 on the proliferation and migration of osteosarcoma cells.3.The difference in lncRNA expression profiles treated and untreated with CCL18 was analyzed by microarray and verified by QRT-PCR.The siRNA was transfected to interfere the expression of UCA1.CCK8,scratch and migration assay were used to detect the influence of si-UCA1 on the proliferation and metastasis of OS cells which treated with CCL18.4.The mRNA microarray was used to analyze the differential expression of transcription factors and to analyze downstream signaling pathways activated by CCL18 through pathway enrichment analysis.The mixture of siRNA were used to interfere the expression of EP300.QRT-PCR was used to observe the expression of UCA1.QRT-PCR and Western Blot were used to detect the expression of ?-catenin and GSK3? in NC and UCA1 knockout osteosarcoma cells treated with CCL18.5.The osteosarcoma nude mice models were constructed to verify the effect of CCL18 on osteosarcoma cell proliferation and migration.6.Immunohistochemistry was used to test the expression of CCL18 in 30 primary breast cancer tissues and bone metastasis tissues.Analysis of the correlation between CCL18 expression and the median time of bone metastases in patients7.Retrospective analysis of 83 breast cancer patients with spinal metastases underwent PVP.The scores of visual analogue scale?Karnofsky performance status?mobility were evaluated 24 h before PVP and 24 h,1 month,3 months,6 months after PVP.RESULTS:1.The CCL18+TAMs count in lung metastases tissue was significantly higher than that in primary tissue.The CCL18 level in serum of osteosarcoma patients was higher than that of healthy people,and CCL18 level was highest in patients with lung metastasis.2.CCL18 promotes the proliferation and metastasis of osteosarcoma cells.3.LncRNA UCA1 and transcription factor EP300 was upregulated in CCL18-treated 143 b osteosarcoma cells,and the downstream Wnt/?-catenin signaling pathway was activated.Silencing UCA1 can weaken the biological effects of CCL18 on osteosarcoma cells.Silencing UCA1 can weaken the mRNA and protein expression of the key molecules of Wnt/?-catenin signaling pathway in osteosarcoma cells treated with CCL18.Silencing EP300 weakened the up-regulation of UCA1 that induced by CCL18.4.The osteosarcoma nude mouse model verified that CCL18 can promote the metastasis of osteosarcoma.5.The expression of CCL18 was up-regulated in breast cancer bone metastasis tissue,and the expression of CCL18 in breast cancer primary tissue was correlated with the median time of bone metastases.6.83 patients were all successfully operated with no serious clinical complications.The pain,quality of life,activity were significantly improved after PVP.CONCLUSION:1.CCL18 promotes osteosarcoma metastasis,which may play this regulatory role through the EP300 / UCA1 / Wnt / ?-catenin pathway.2.The expression of CCL18 is up-regulated in breast cancer bone metastasis tissue and was closely related to median time of bone metastases,suggesting that CCL18 plays an important role in the process of bone metastasis of breast cancer.3.PVP has a good clinical effect on breast cancer patients with vertebral body metastases,which can significantly relieve pain,improve quality of life and has good safety.