Eps8 Gene On The Proliferation Of Glioma Cells

As one of the most common malignant brain tumor, glioma harms human life and health seriously. But its pathogenesis is not yet fully understood.Most reports show that Eps8 is related to the rapid cell growth and cell migration in many cancers.However, the role of Eps8 in glioma has not been studied until now.Immunohistochemical analysis of 32 glioma tissues and 3 adjacent normal brain tissues, the results showed that Eps8 was highly expressed in 17 samples of IV glioma, moderately expressed in 14 samples of IV glioma, but lowly expressed in only 1 sample ofⅢglioma, none in adjacent normal brain tissues, which indicated that Eps8 has a high expression level in most gliomas and the expression levels of Eps8 may be related to malignanty of glioma.Besides, we detected the expression levels of Eps8 in human glioma cells by Western blotting, and as a result, we found Eps8 express in all of these cell lines, but lower in U251 than those in SHG44 and A172. It suggested that Eps8 has different expression levels in different cell lines.In order to further study the role of Eps8 in glioma development,we constructed eukaryotic expression vector pEGFP-C3-Eps8 by the sub-cloning.Then,we transfected it into U251 cells with low expression of Eps8 by lipofectamine TM 2000.After screening and expanding cells by 400μg/mL G418,we successfully established U251 cell line stably expressing Eps8, which was demonstrated by fluorescence localization assays and western blotting.Cell survival and MTT assays confirmed that Eps8 can increase cell proliferation and improve the relative activity of U251 cells. At the same time,colony formation assay showed that Eps8 can improve the ability of U251 cells to form colonies. These results indicated that Eps8 can facilitate U251 cell growth and proliferation.We also found that the number of U251-GFP/Eps8 cells in G1 phase was less than the other two control groups by analyzing cell cycle with Flow cytometry. Moreover, the number of the S phase cells was more than control groups, which showed that Eps8 can promote U251 cells from G1 phase to S phase. Finally,in order to search the molecule mechanism that Eps8 can promote U251 cell proliferation,we found that Eps8 overexpression can enhanceβ-catenin expression and suppress caspase 3 expression. These suggested Eps8 can increase glioma proliferation through activating Wnt/β-catenin signal pathway and inhibiting apoptosis marker gene caspase 3.In conclusion, these results support that Eps8 acts as an oncogene to promote cell proliferation.Eps8 plays an important role in the development of glioma.It may be a potential therapeutic target in clinical diagnosis and glioma treatment.