The Study On The Expression Of BNIP3 In Medulloblastoma Cell

Objective BNIP3(Bcl-2/adenovirus E1B 19 kDa-interacting protein 3) is a type of mitochondrial protein,belong to apoptosis Bcl-2 protein family members, promote cell apoptosis. Research shows that, in the development of many tumors,BNIP3 plays an important role.Medulloblastoma is a high-degree malignant of intracranial neuroepithelial tumors,and little was reported about the expression of BNIP3 in medulloblastoma and so was its function of tumor development. The aim of this article is to detect the expression of BNIP3 in human medulloblastoma cell at the levels of proteins and genes, and used the remover of DNA methylation on the medulloblastoma cell, to investigate the relevance of overexpression of BNIP3 ,and the occurrence,development of human medulloblastoma;and investigate its relationship of induce the medulloblastoma cell apoptosis, discusses the BNIP3 whether can be considered as a treatment targets of medulloblastoma.Methods (1)Cultivated medulloblastoma cells(Med-341),and observed the growth of cells; ten normal cerebellum tissues (non-tumor cerebellum tissues) were obtained from Neurosurgery Department of the First Affiliated Hospital of Anhui Medical Universityas as normal control from surgical resections of trauma patients, for whom a partial resection of normal cerebellun tissue was required as decompression treatment for their severe head injuries to reduce increased intracranial pressure. The SP immunohistochemistry method was use to detect the expression of BNIP3 in medulloblastoma cell and 10 cases of non-tumor cerebellum tissue samples.(2) The medulloblastoma cell was collected;five fresh non-tumor cerebellum tissues were collected from the First Affiliated Hospital of Anhui Medical University during the operation of neurosurgery. All tissues were diagnosised as non-tumor cerebellum by two senior pathologists. The expression of BNIP3 mRNA was examined by semi-quantity reverse transcription-polymerase chain reaction (RT-RT-PCR) in medulloblastoma cell and 5 non-tumor cerebellum tissues,Hep-2 cell was chosen as positive control.(3) The medulloblastoma cell was cultivated in bottles, all was gived different concentration of remover of DNA methylation(5-Aza-2-Deoxycytidine) for 7 days , compared with the medulloblastoma cell with nothing.Then cultivated the cell in 96-bore borade, the influence of cell generation was examined by MTT.Rusluts (1)The expression of BNIP3 was positive by immunohistochemical and RT-PCR in medulloblastoma cell;in non-tumor cerebellum tissue samples, no expression of BNIP3 was detected.(2)In the experimentation of observation the influence of remover of DNA methylation (5-Aza-2-Deoxycytidine),we were discoverd that the cell was depressed by the concentration of 5-aza-cytosine deoxyriboside above 1.0μmol/L.This cued that remover of DNA methylation ( 5-Aza-2-Deoxycytidine ) may induse the expreesion of BNIP3.The inhibition rate up to 38% by the 5-Aza-2-Deoxycytidine of 1.0μmol/L.And this inhibitory activity had dose-response relationship in certain concentration.Conclusion (1)The level of BNIP3 in medulloblastoma cell is higher than in non-tumor cerebellun tissue.There was a relationship between overexpression of BNIP3 and the development of human medulloblastoma.(2)Remover of DNA methylation(5-Aza-2-Deoxycytidine)could inhibit proliferation of medulloblastoma cell,and in certain concentration with concentration increases, its effect was enhanced. This cued that remover of DNA methylation (5-Aza-2-Deoxycytidine)could induce the expression of BNIP3,and BNIP3 could be one treatment targets of medulloblastoma.