传代次数对人胶质瘤U87细胞系生物学特性的影响 postprint

Abstract: Objective To unravel the differences of biological characteristics and the uderlying molecular mechanisms betweendifferent passages of U87 glioblastoma cell line.Methods U87 (I) and U87 (II), with less or more passage number respectively,were established examined separately by MTT assay, transwell chamber assay, boyden chamber assay. Western blot was usedto analyze the expression of CTHRC1, FOXM1, PLOD2, MMP9, TGF-β, E-cadherin, Slug, Snail, Vimentin, PI3K, p-PI3K, Aktand p-Akt separately in these two types of cells.Results Compared to U87 (II), U87 (I) was more prone to form tubules andshowed more invasiveness, while there were no differences between U87 (I) and U87 (II) in proliferation and migration. Theexpression of markers of EMT varies in these two types of cells, with more expression of E-cadherin and Slug and lessexpression of Vimentin and Snail in U87 (II). Moreover, U87 (II) was found to have greater amount of Akt and p-Akt, but havesmaller amount of p-PI3k. No significant difference was found on the expression of PI3K between these cells. Additionally, theexpression of PLO2, CTHRC1 and MMP9 were higher in U87 (I) than that in U87 (II), while the expression of TGF-β in U87 (I)was lower than that in the latter one.Conclusion As the passage number increased, U87 cell lines exhibited changes ininvasiveness as well as some oncogenic genes’ expressions. It may lead to the different results during different periods, makingthe results inconvincible. Together, our results showed that, to lessen the influence of passage numbers on cells’ biologicalcharacteristics, scientists should use cells with same passage numbers and finish the experiments as quickly as they can.