ABSTRACT
OBJECTIVE@#To investigate the effect of JAG1 on the malignant phenotype of triple-negative breast cancer (TNBC) and its role in angiogenesis in breast cancer microenvironment.@*METHODS@#The expressions of Notch molecules were detected in human TNBC 231 and 231B cells using RT-qPCR. Five female nude mice were inoculated with 231 cells and another 5 with 231B cells into the mammary fat pads, and 4-6 weeks later, the tumors were collected for immunohistochemical and immunofluorescence tests. 231 cells and 231B cells were treated with recombinant JAG (rJAG) protein and DAPT, respectively, and changes in their malignant phenotypes were assessed using CCK-8 assay, Hoechst 33258 staining, wound healing assay, Transwell chamber assay and endothelial cell adhesion assay. Western blotting was used to detect the changes in the expressions of proteins related with the malignant phenotypes of 231 and 231B cells. The effects of conditioned medium (CM) derived from untreated 231 and 231 B cells, rJAG1-treated 231 cells and DAPT-treated 231B cells on proliferation and tube formation ability of cultured human umbilical vein endothelial cells (HUVECs) were evaluated using CCK-8 assay and tube-forming assay.@*RESULTS@#The expression of JAG1 was higher in 231B cells than in 231 cells (P < 0.05). Tumor 231B showed higher expression of VEGFA and CD31. Compared with 231-Blank group, the migration, invasion and adhesion of 231 cells in 231-rJAG1 were significantly enhanced (P < 0.05). Protein levels of Twist1 and Snail increased (P < 0.01), anti-apoptotic protein Bcl-2 increased (P < 0.05), while DAPT inhibited the related phenomena and indicators of 231B. The 231-rJAG1-CM increased the cell number and tubule number of HUVEC (P < 0.05).@*CONCLUSION@#JAG1 may affect the malignant phenotype of TNBC and promote angiogenesis in the tumor microenvironment.
Subject(s)
Animals , Female , Humans , Mice , Cell Line, Tumor , Cell Movement , Cell Proliferation , Culture Media, Conditioned , Human Umbilical Vein Endothelial Cells/metabolism , Jagged-1 Protein/metabolism , Mice, Nude , Neovascularization, Pathologic/metabolism , Platelet Aggregation Inhibitors , Sincalide/metabolism , Triple Negative Breast Neoplasms/metabolism , Tumor MicroenvironmentABSTRACT
[Abstract] Objective To study the clinical diagnosis and treatment of parkinsonism (PDS) with freezing of gait (FoG), so as to provide clues to delay the progress of the symptom. Methods A prospective study was designed. The outpatients of PDS with the main complaint of FoG were included and followed up for 2-6 years in the Department of Neurology, Changzheng Hospital, Naval Medical University (Second Military Medical University) from Nov. 2010 to Jan. 2016. The patients were given L-dopa first, and then antidepressants and other therapies (including other medication and surgery) were given if the previous treatments were not effective. The motor function of patients was evaluated by Hoehn-Yahr staging scale and the second and third part of the unified Parkinson disease rating scale (UPDRS); the general mental, behavior and emotional state were evaluated by the first part of UPDRS; the cognition was evaluated by minimum mental state examination (MMSE); depression and anxiety were evaluated by 17-item Hamilton depression scale (HAMD-17) and Hamilton anxiety scale (HAMA); and the severity of FoG was evaluated by the timed up and go test (TUGT). Results Six of the 15 cases with FoG were diagnosed as Parkinson disease (PD), and 9 had other disorders (2 with progressive supranuclear palsy, 3 with primary progressive FoG, 1 with frontotemporal dementia, 1 with vascular PDS, 1 with drug-induced PDS, and 1 with unknown-cause PDS). There were no significant differences in age, gender, severity of symptom or mental state (Hoehn-Yahr stage, UPDRS- score, UPDRS-Ⅱ score, UPDRS-III score, MMSE score, HAMD-17 score, HAMA score and TUGT time) between PD group and non-PD group (all P0.05). At the baseline, the FoG duration of PD patients ([7.50±2.66] years) was longer than that of non-PD patients ([2.56±0.88] years, P0.01). After treatment with increasing dose of L-dopa, 4 PD patients were improved while non-PD patients had no responses (4/6 vs 0/9, P＝0.01). Conclusion The causes of PDS with FoG are heterogeneous. The duration of FoG is helpful for diagnosis of idiopathic PD, while the severity of FoG has little value for etiological analysis. Increasing the dose of L-dopa is effective for FoG in advanced PD, while it has uncertain effect for FoG of other reasons.