MicroRNA-27a-5p Inhibits Proliferation, Migration, and Invasion and Promotes Apoptosis of Wilms' Tumor Cell by Targeting PBOV1.

Wilms' tumor is the most common type of renal tumor in children. MicroRNAs (miRNAs) are small noncoding RNAs that play crucial regulatory roles in tumorigenesis. We aimed to study the expression profile and function of miR-27a-5p in Wilms' tumor. miR-27a-5p expression was downregulated in human Wilms' tumor tissues. Functionally, overexpression of miR-27a-5p promoted cell apoptosis of Wilms' tumor cells. Furthermore, upregulated miR-27a-5p delayed xenograft Wilms' tumor tumorigenesis in vivo. Bioinformatics analysis predicted that miR-27a-5p directly targeted the 3'-untranslated region (3'-UTR) of PBOV1, and luciferase reporter assay confirmed the interaction between miR-27a-5p and PBOV1. The function of PBOV1 in Wilms' tumor was evaluated in vitro, and knockdown of PBOV1 dampened cell migration. In addition, overexpression of PBOV1 antagonized the tumor-suppressive effect of miR-27a-5p in Wilms' tumor cells. Collectively, our findings reveal the regulatory axis of miR-27a-5p/PBOV1 in Wilms' tumor, and miR-27a-5p might serve as a novel therapeutic target in Wilms' tumor.

Propranolol induces regression of hemangioma cells via the down-regulation of the PI3K/Akt/eNOS/VEGF pathway.

BACKGROUND: Infantile hemangioma (IH) is a benign vascular neoplasm resulting from the abnormal proliferation of endothelial cells and pericytes in infants. Propranolol, a non-selective ß-adrenergic blocker, has recently emerged as an effective therapy for IH, causing regression. However, its potential therapeutic mechanism remains largely unknown. PROCEDURE: An XPTS-1 cell line was established by isolating hemangioma-derived endothelial cells (HemECs) from a specimen of human proliferating IH. Flow cytometer assay was performed to assess the effect of propranolol on cell cycle distribution. Western blot was employed to determine changes of protein expression. Matrigel invasion and tube formation assays were used to measure invasion ability and tube formation ability, respectively. Commercial kits were employed to quantify NO and VEGF levels. RESULTS: Propranolol blocked norepinephrine-induced HemECs cell cycle progression as well as the expression of cyclin A2 and cyclin D2; whereas p21 and p27 proteins were altered conversely. Propranolol inhibited norepinephrine-induced cell invasion by reducing the expression of MMP-9, VEGF, and p-cofilin. NO and VEGF release induced by norepinephrine was decreased by propranolol pretreatment, coincident with alterations in the phosphorylation of Akt, eNOS, and VEGFR-2. Tube formation ability and subsequent levels of NO and VEGF elevated by norepinephrine were distinctively counteracted in HemECs. CONCLUSIONS: The current study demonstrated the antiangiogenic properties of propranolol in vitro and that the drug was able to induce the regression of hemangioma cells via the inhibition of cell cycle progression, invasion, and tube formation, concomitantly with decreased NO and VEGF levels through the down-regulation of the PI3K/Akt/eNOS/VEGF pathway.

[Therapeutic mechanism of bleomycin A5 on infancy hemangioma: an experimental study].

OBJECTIVE: To investigate the therapeutic mechanism of Bleomycin A5 on infancy hemangioma. METHODS: After intralesional injection of Bleomycin A5 into the tumor of animal model of infancy hemangioma, the variation of tumor form was and the variation of tumor structure were observed using light microscope and electron microscope, the variation of tumor gene expression spectra was also tested by DNA microarray technique. RESULTS: After treatment, the tumor gradually shrunk, hardened, disappeared one month later. The tumor lost appearance of infancy hemangioma and replaced by lamellar collagen fibers and cellular nucleus scattered in the fibers, and almost all cells were necrotic and dissolved. Under electron microscope, only large stretches of dissolved cell could be seen without intact cells and blood vessels, but apoptotic cells and bodies could also be found. The results of DNA microarray analysis showed that 9 genes associated with apoptosis (murine double minute 2, heat-labile enterotoxin B subunit, lymphotoxin B receptor, tumor necrosis factor ligand superfamily 7, tumor necrosis factor receptor superfamily 21, tumor necrosis factor receptor superfamily 1A, myeloid cell leukemia-1, caspase3), 13 genes associated with cell proliferation and cell cycle (cell division cycle27, cell division cycle37, CDC28 protein kinase 1B, cycling B1, cullin 2, cullin 3, cullin 4A, growth arrest and DNA damage-inducible 45A, meiotic recombination 11 homolog B, forkhead box M1, minichromosome maintenance 7, antigen identified by monoclonal antibody ki 67, proliferating cell nuclear antigen), and 11 genes associated with cellular stress and toxic reaction (glutathione peroxidase 1, metallothioneins, superoxide dismutase-1, heat shock protein A1A, heat shock protein A2, heat shock protein A4, heat shock protein A5, heat shock protein 9B, heat shock protein CA, macrophage migration inhibitory factor, plasminogen activator inhibitor)were up or down regulated more than 2 folds in tumors treated with Bleomycin A5 compared with controls. CONCLUSIONS: The therapeutic effect of Bleomycin A5 on infancy hemangioma is the synthetic results of multiple factors. Bleomycin A5 could not only induce apoptosis and inhibit cell proliferation, but also depressed the ability of cell stress and toxic reaction.

[Effect of bleomycin A5 on the hemangioma-derived endothelial cell line XTPS-1].

OBJECTIVE: To investigate the effect of bleomycin A5 on the hemangioma-derived endothelial cell line XTPS-1. METHODS: Hemangioma-derived endothelial cell line XTPS-1 was cultured with different concentration of bleomycin A5 (1000, 100, 10, 1, 0 mg/L), and then the survival rate was measured by methyl thiazolyl terazolium (MTT), the variation of cell morphology was observed using inverted phase contrast microscope and electron microscope, the variation of cell cycle and apoptosis rate were measured using flow cytometry. RESULTS: After 24 hours culture the cell survival rate was (92.96 ± 3.66)% and (99.86 ± 0.12)% in lower saturation group (10 and 1 mg/L), but (34.08 ± 3.11)% and (43.28 ± 2.88)% in higher saturation group (1000 and 100 mg/L). The difference between them was more significant (P < 0.01). Lower saturation of bleomycin A5 (10 and 1 mg/L)could induce apoptosis but had almost no cytotoxic effect. Higher saturation of bleomycin A5 (1000 and 100 mg/L) not only induced apoptosis, but also had strong cytotoxic effect, which was concentration dependent. CONCLUSIONS: bleomycin A5 could induce apoptosis, inhibit cell proliferation and has direct cytotoxic effect.

[Establishment of human infancy hemangioma-derived endothelial cell line XPTS-1 and animal model of human infancy hemangioma].

OBJECTIVE: To establish an immortalized human infancy hemangioma-derived endothelial cell line (HemEC) and animal model of human infancy hemangioma. METHODS: Hemangioma-derived endothelial cells from specimen of human infancy hemangioma were cultured in vitro and monocloed, and then its growth curve was made, karyomorphism of chromosome analyzed, morphologic characteristics observe, factor VIII related antigen identified by immunohistochemical method.Vascular endothelial growth factor receptor 2 (VEGFR-2) was detected by flow cytometry. HemEC were inoculated subcutaneously in athymic mouse to establish animal model of infancy hemangioma. The animal model was observed closely and its pathological characteristic was also studied. RESULTS: The cultural cells grew active, and immortalized spontaneously when they were subcultured on sixteenth generation. This cell line was cultivated for more than 70 times within one year and in good condition after freezing and resuscitating once and again, and had the morphologic character of HemEC. The cell population doubling time was 22 h. Factor VIII and VEGFR-2 were expressed positively. Karyo type analysis of the cell line showed abnormal diploid with the modal chromosomal number varying between diploid and triploid. The cell line was then named XPTS-1. The animal model of infancy hemangioma was successfully established and its character of histopathology was similar with that of infancy hemangioma. CONCLUSIONS: The cell line of HemEC was successfully established and immortalized spontaneously, and had the morphologic and biological character of HemEC. The animal model of infancy hemangioma was successfully established and showed the character of histopathology similar with that of infancy hemangioma.