RESUMO
Objective:To investigate mechanisms underlying the signal crosstalk of VEGF-IL-6-STAT3 between cutaneous melanoma cells and vascular endothelial cells.Methods:EC-304 vascular endothelial cells were divided into 3 groups: control group cultured in conventional endothelial cell-conditioned medium, vascular endothelial growth factor (VEGF) group cultured in endothelial cell-conditioned medium containing 50 μg/L VEGF 165, A375 co-culture group co-cultured with a melanoma cell line A375. After 24-, 48- and 72-hour treatment, the culture medium was collected, and enzyme-linked immunosorbent assay was performed to detect the level of interleukin-6 (IL-6) . Cultured A375 cells were divided into 4 groups: control group receiving conventional culture in Dulbecco′s modified Eagle′s medium (DMEM) , A375+ EC-304 group co-cultured with EC-304 cells, A375+ EC-304+ IL-6 group co-cultured with EC-304 cells in DMEM containing 50 μg/L IL-6 (an agonist of the signal transducer and activator of transcription-3 [STAT3] pathway) , A375+ EC-304+ JSI-124 group co-cultured with EC-304 cells in DMEM containing 1 μmol/L JSI-124 (a STAT3 pathway inhibitor) . After 24-, 48- and 72-hour treatment, cells were collected, and Western blot analysis, cell counting kit-8 (CCK8) assay and Transwell invasion assay were performed to determine the protein expression of STAT3 and phosphorylated (p) -STAT3, cellular proliferative activity and invasive activity, respectively. Two-way analysis of variance and t test were used for statistical analysis. Results:The level of IL-6 significantly increased in the culture medium of EC-304 cells in the VEGF group and A375 co-culture group compared with the control group ( FVEGF = 29.63, P < 0.001; FA375 = 11.09, P = 0.020) . Compared with the control group, the A375+ EC-304 group showed significantly enhanced protein expression of p-STAT3 in A375 cells ( P < 0.001) , increased cell activity ( P < 0.001) , and increased number of invasive cells (152.66 ± 16.04 vs. 86.13 ± 7.24, t= 4.43, P < 0.001) ; compared with the A375+ EC-304 group, the A375+ EC-304+ IL-6 group showed significantly increased protein expression of p-STAT3 ( P < 0.001) , enhanced cell activity ( P < 0.001) , and increased number of invasive cells (187.34 ± 14.38, t= 2.17, P < 0.001) ; compared with the A375+ EC-304 group, the A375+ EC-304+ JSI-124 group showed significantly decreased protein expression of p-STAT3 ( P < 0.001) , decreased cell activity ( P < 0.001) , and decreased number of invasive cells (124.92 ± 8.72, t=-1.86, P < 0.001) . Conclusion:There is a signal crosstalk of VEGF-IL-6-STAT3 between cutaneous melanoma cells and vascular endothelial cells, which may play an important role in the proliferation and invasion of A375 cells.
RESUMO
Objective To summarize our experience in microsurgical treatment of dumbbell tumors. Methods The clinical features,surgical approaches,operative techniques and surgical results of 21 cases of dumbbell tumors were analyzed retrospectively.Four surgical approaches were adopted according to the size and location of tumors,including,posterior midline approach(in 7 cases),modified far lateral approach(in 5 cases),posterior midline supplied by anterior cervical approach(in 2 cases)and paramidline approach(in 7 cases). Results In this series,10 cases of tumors were situated in the cervical,6 in thoracic,4 in lumbar and 1 in sacral segment of the spine.Total removal were achieved in 95% (20/21),and subtotal removal in 5%(1/21).Postoperative pathological diagnosis revealed 15 neurinomas,5 spinal meningiomas and 1 metastatic adenocarcinoma.14 cases of meningiomas and 1 metastatic adenocarcinoma were followed up for 1 to 48 months.11 of the 14 patients recovered completely,with 3 of them living daily life independently,4 maintainly radical hypoesthesia,and one having decubital ulcer unhealed.No postoperative malformation and dysfunction of the disposed spine occurred in these patients.Magnetic resonance imaging scans in 11 of the 14 follow-up cases revealed no tumor recurrence,of which inhanced MRI scans were performed in 8 cases. Conclusions dumbbell tumors should be treated by one-stage operation.Microsurgical technique could improve the rate of total removal,and decrease postoperative complications.