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The Expression and Relationship of NANOG ,CD133 and GFAP in Gliomas

Author: LiuYuHai
Tutor: NiuChaoShi；FuXianMing
School: Anhui Medical University,
Course: Surgery
Keywords: NANOG Immunohistochemical double CD133 Cancer stem cells U87
CLC: R739.41
Type: Master's thesis
Year: 2011
Downloads: 58
Quote: 0
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Abstract

Objective NANOG, CD133, GFAP glioma tissue in vivo coexpression explore NANOG gene and CD133, GFAP expression of the relationship between; cultured cell lines U87 identified cancer stem cells, cell lines in vitro study on the level of NANOG and CD133 coexpression relationship analysis NANOG in gliomas in the development of the role played. Methods (1) double immunohistochemical assay NANOG, CD133, GFAP in 60 cases of glioma tissue expression to calculate NANOG ~ cells, CD133 ~ cells, GFAP ~ cells, NANOG ~ / CD133 ~ cells and NANOG ~ / GFAP ~ the percentage of cells and the expression of the positive indicators were statistically analyzed. (2) The method of serum deprivation in U87 glioma stem cells in culture, tumor cells were observed the formation of the ball; ball of the tumor inoculated in the serum medium in vitro differentiation observed characteristics; immunofluorescence of cells surviving the serum deprivation and proliferation of cells and the formation of tumors ball differentiated cells. (3) double immunocytochemical detection of tumor stem cells and CD133 ball NANOG co-expression, and to analyze the relationship between the expression. Results (a) levels in different pathological human glioma NANOG ~, CD133 ~ and GFAP ~ cell percentage difference was significant, and the expression of CD133 ~ NANOG ~ expression was positively correlated (r = 0.904, P lt; 0.001), NANOG ~ expression of GFAP ~ expression was negatively correlated (r = -0.907, P lt; 0.001). With the increase of pathological grade, NANOG ~ / CD133 ~ cell percentage difference was significant (F = 560.805, P lt; 0.001), NANOG ~ / GFAP ~ cells (F = 0.355, P gt; 0.1) percentage difference was not statistically significance, and NANOG ~ / CD133 ~ CD133 ~ expression was positively correlated (r = 0.823, P lt; 0.001). (2) using serum deprivation method was successfully screened out of the expression of CD133 cancer stem cells, and can form a tumor proliferation ball; ball can be more tumor differentiation, progeny table GFAP and NSE; dual immunocytochemical staining NANOG and CD133 CPC in cancer stem cell spheres expressed in stem cell differentiation ball around in semi-CD133 cells expressing only without express NANOG. Conclusions (1) at different levels showed positive glioma NANOG expression and glioma stem cell markers CD133 significant correlation between, and brain tumors CD133, NANOG positive cell ratio and the degree of malignancy with CD133 expression in tumor tissue also expressed NANOG, and there are co-expressed on the cell level. Pathology in glioma grading, NANOG ~ expression and GFAP ~ expression levels were negatively correlated. NANOG expression of tissue, GFAP expression levels are relatively low, most of the cells did not NANOG and GFAP co-expression of different pathological levels NANOG ~ / GFAP ~ no statistically significant difference in the percentage of cells (2) U87 cell lines exist self-renewal and multi-differentiation capacity glioma stem cells; initial serum deprivation method used successfully from U87 glioma cell lines were isolated and screened glioma stem cell proliferation and tumor formation balls. Immunohistochemical double discovery NANOG and CD133 in the level of expression of stem cells, NANOG in cancer stem cells with high expression, is not expressed in differentiated cells, stem cells may be maintained in an undifferentiated state plays a very important role.

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