| Purpose: to investigate the changes of moleculal markers in cultured skin stem cells exposure to UVB in vitro.Methods: The skin stem cells were isolated and cultured according to their adhered ability and identified by K15 andβ-integrin expression. The skin stem cells were exposured in the UVB irradiation. The UVB′s peak value was 305nm and the dosages was 10 mJ/cm2. The morphological changes of skin stem cells after UVB irradiation (experimental group) or not (control group) were observed. The changes of CD34,beta-catenin,P53 of skin stem cells were detected by immunohistochemisty and compared in UVB exposure group or not.Results:①The skin stem cells were successfully separated and cultured. They expressed characteristic skin stem cell marker K15 andβintegrin.②Cells of control group had a dense density.They were round or polygon. the cells were found to have high N/C ratios and the dividing cells can be found. The morphology was clear,and the cytoplasm was well-distributed. In contrast, the skin stem cells in experimental group, grew slowly the cell morphology was deformities and anomalous, vacuoles were detected in cytoplasm. The cells were found to have low N/C ratios.a proportion of cells were karyopyknosis or apoptosis.③In the control group of skin stem cells, beta-Catenin were expressed premodinantly in the cell membrane or cytoplasm and its membrane and nucleus stained positive rates were 64.74%, 8.4%.;P53 were expressed mainly in the cytoplasm and their nuclei stained positive rate were 6.9%. In contrast,in the experimental group of skin stem cells,beta-Catenin expression mainly in the cytoplasm and the nucleus, the cell membrane and stained positive rates were 64.74% ,0. P53 expression mainly in the nucleus, the nucleus staining was 100%. CD34 was not expressed in both cases.④K15 andβ1 integrin expression in the skin stem cells of control group were in the cytoplasm,and the expression of K15 andβ1 integrin were all disappeared after UVB irradiationConclusion:1.Using rapid adhesion method could separate skin stem cells from keratinocytes. They expressed characteristic skin stem cell marker K15 andβintegrin. 2.UVB can promote beta-catenin to accumulate in cytoplasm and immigrate to nucleus.3.P53 were located in the cytoplasm and immigrated to nucleus when exposed in UVB irradiation.4.The expression of K15 andβ1 integrin in the skin stem cells were all disappeared after UVB irradiation... |
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