Construction and identification of U87 glioblastoma cell strain with a stable low expression of BAG3
LI Ji-qiang
YANG Ji-an
SHAO Ling-min
WU Ting-feng
LIU Bao-hui
CHEN Qian-xue
Abstract：Objective To construct and identify glioblastoma U87 cell strain with a stable low expression of BAG3. Methods The plasmids of pEGFP-BAG3-shRNA were extracted by RNA interference technique and then were sequence by gene sequencing instrument ABI3130. The plasmids of pEGFP-BAG3-shRNA which met the sequence identical outcomes were transfected into U87 glioblastoma cell strain. The transfected U87 glioblastoma cell strains were sifted out by puromycin and then were identified by RT-PCR, western-blotting and immunofluorescence techniques. Results The sequencing outcomes showed that pEGFP-BAG3-shRNA plasmid contained objective gene interfering sequence. The percentage of tranfected U87 glioblastoma in shBAG3 group and shCtrl group were 85%and 93%respectively. The levels of BAG3 mRNA and protein expressions were significantly lower in shBAG3 group than those in shCtrl group. Conclusion U87 glioblastoma cell strain constructed by the present study can stably express BAG3 at a low level and may be used for the further research of the role of BAG3 in the gliobalstoma and its associated signaling pathways.
Keywords：GlioblastomaU87 cell strainBAG3RNA interference
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 353-356 )