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Objective To investigate the effect of miR-181c-5p on vasculogenic mimicry (VM) in ovarian cancer stem-like cells (OCS-LCs) and its mechanism. Methods OVCAR3 cells were induced into OCS-LCs by serum-free suspension culture. OVCAR3 cells were divided into groups A, B, and C and were transfected with NC-miR-181c-5p, siRNA-miR-181c-5p, and pRNA-miR-181c-5p, respectively. The sphere-forming experiment was used to assess the sphere formation ability of groups A, B, and C. RT-qPCR was used to measure the relative expression level of miR-181c-5p in cells of groups A, B, and C, and Western blot was used to measure the relative protein expression levels of Oct-4, Nanog, HIF-1α, and VEGF in cells of groups A, B, and C. CCK-8 assay was used to measure the viability of cells in groups A, B, and C, and the three-dimensional culture experiment was used to measure the angiogenesis rate of groups A, B, and C. Results OVCAR3 cells were successfully induced into OCS-LCs. RT-qPCR showed that group B had a significantly lower relative expression level of miR-181c-5p than group A, and group C had a significantly higher relative expression level than group A (t=2.25,8.68,P

Objective To investigate effects of alpinetin (APT) on angiogenesis in knee osteoarthritis (KOA) rats by regulating vascular endothelial growth factor/sphingosine kinase 1/sphingosine 1 phosphate (VEGF/SphK1/S1P) signaling pathway. Methods The KOA rat model was established by Videman method. Ninety rats were grouped into the control group, the model group, the low-dose kaempferol group (L-APT group), the high-dose kaempferol group (H-APT group), the high-dose kaempferol group+lentivirus negative control group (APT+NC group) and high-dose kaempferol+overexpression of SphK1 lentivirus group (APT+SphK1 group), with 15 rats in each group. Pathological changes of cartilage tissue in rats were observed by HE staining. Contents of IL-1β, TNF- α, IL-6 and MMP-13 in cartilage tissue were measured by enzyme linked immunosorbent assay. Chondrocyte apoptosis of cartilage tissue cells was detected by TUNEL. VEGF and CD31 protein positive expression levels were detected by immunohistochemistry assay. The p-VEGFR2, VEGFR2, SphK1 and S1P protein levels were detected by Western blot assay. Results Rats in the model group showed pathological damage. Compared with the control group, the apoptosis rate, IL-1b, TNF-a, IL-6, MMP-13 levels, VEGF positive expression, CD31 positive expression, p-VEGFR2, SphK1 and S1P protein expression levels were increased in the model group (P＜ 0.05). Compared with the model group, the pathological damage was obviously reduced in the L-APT group and the M-APT group, and cell apoptosis rate, IL-1β, TNF-α, IL-6, MMP-13 levels, VEGF positive expression, CD31 positive expression, p-VEGFR2, SphK1 and S1P protein expression levels were obviously reduced (P＜0.05). Compared with the APT+NC group, the pathological injury of cartilage tissue increased in the APT+SphK1 group, cell apoptosis rate, IL-1β, TNF-α, IL-6, MMP-13 levels, VEGF positive expression, CD31 positive expression, p-VEGFR2, SphK1 and S1P protein expression levels were obviously increased (P＜0.05). Conclusion APT inhibits angiogenesis in knee osteoarthritis rats by inhibiting the VEGF/SphK1/S1P signaling pathway. [ABSTRACT FROM AUTHOR]