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The Effect of mTOR/p70S6K Signal Pathways in Children Hemangioma

Author: LvXin
Tutor: YuSong
School: Zunyi Medical College,
Course: Pediatric Surgery
Keywords: Hemangioma Vascular endothelial cells Cell culture mTOR p70S6K Cell cycle Rapamycin
CLC: R732.2
Type: Master's thesis
Year: 2010
Downloads: 57
Quote: 0
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Abstract


Purpose to explore mTOR/p70S6K signaling pathways in children with hemangiomas occur, process development and subsided. Method 1. Collect the Affiliated Hospital of Zunyi Medical College, 2005 -2009 without other treatment, simple surgical excision by pathological diagnosis of hemangioma specimens of 31 cases, combined Mulliken classification and proliferating cell nuclear antigen (PCNA) expression of hemangioma classification and staging, immunohistochemical detection and proliferating hemangiomas and involuting hemangioma tissue of mTOR, p70S6K-α expression level. Using tissue explant culture of endothelial cells to be grown into a monolayer cells after passaging, the establishment of the Department of hemangioma vascular endothelial cells, to observe the proliferation characteristics and identification Ⅷ coagulation factor related antigen. Synchronization cultured endothelial cells, changes Western blot assay p70S6K-α expression levels in endothelial cells, the same period the hemangioma vascular endothelial cell cycle by flow cytometry.; MTOR, p70S6K-α expression levels correlation with hemangioma vascular endothelial cell cycle distribution and apoptosis. 3 When the cells in the logarithmic phase, for serum-free medium and incubated 48h cell synchronization, and then were added 0, rapamycin 10nmol / L, and continue for 24 hours, cells were collected by cell scraper, detection of endothelial cells mTOR p70S6K,-α the expression level, while detecting vascular tumor vascular endothelial cell cycle distribution. 1.18 cases of proliferative phase hemangioma mTOR, p70S6K-α integral optical density were 6336.48 ± 1655.89,588.72 ± 223.87; 13 cases of the resolution phase hemangioma mTOR, p70S6K-α integral optical density were 846.22 ± 297.09,3235.64 ± 947.77, ; hemangioma proliferation phase the p70S6K-α of integrated optical density was significantly lower than the resolution phase, the difference was significant (P lt; 0.01); the vascular the tumor proliferative phase mTOR integrated optical density was significantly higher than the resolution phase, the difference was statistically significant (P lt; 0.01). 2. Cultured endothelial cells in the four days after adherent growth, only a week after a small amount of cell spreading adherent growth, growth and division after two weeks, the cells began rapid growth after three weeks, four weeks after cell is covered with the bottom of the plate. Observed with a phase contrast inverted microscope, it is found that the morphology of the isolated endothelial cells goes to mesothelial cells. Cultured cells Ⅷ clotting factor associated antigen expression is positive. 3.mTOR higher levels of protein expression, the p70S6K-α protein expression level is lower, in the G0/G1 phase of the cell less (55.95 ± 4.38)%; mTOR protein expression levels, the p70S6K-α protein expression levels, in the Go / Gl phase cells increased significantly (77.86 ± 8.18)%. Rapamycin hemangioma vascular endothelial cells cultured in vitro for 24 hours, the cell cycle becomes arrested in the G1 phase, the difference was significant (P lt; 0.01) before and after the action of rapamycin; mTOR protein expression levels (P lt; 0.01); p70S6K-α protein levels (P lt; 0.01). Conclusion 1. Highly expressed in the proliferative phase of hemangioma mTOR may lead to the activation of p70S6K-α phosphorylation promote hemangioma vascular endothelial cells from G0/G1 into S phase, hemangioma was a rapid proliferation of state. Involuting hemangioma, in the role of some kind of mechanism, mTOR expression was reduced, inhibited the phosphorylation of p70S6K-α, and cell cycle arrest in G0/G1 phase, thereby promoting vascular tumor vascular endothelial cell apoptosis, hemangioma showed spontaneous regression of the trend. Hemangioma endothelial cells cultured in vitro, mTOR high expression led to of p70S6K activation of phosphorylation, promote vascular tumor vascular endothelial cells from G0/G1 into S phase, the rapid proliferation of vascular endothelial cells; decreased expression of mTOR, inhibited the phosphorylation of p70S6K and cell cycle arrest in G0/G1 phase, thus contributing to the vascular endothelial cell apoptosis. Rapamycin hemangioma endothelial cells cultured in vitro stranded at the G0/G1.

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