Haptoglobin gene expression in human glioblastoma cell lines.
Increases in cerebrospinal fluid (CSF) levels of the acute phase protein haptoglobin (Hp) occur in central nervous system (CNS) disorders such as Alzheimer's disease. To establish if Hp CSF level increases can be associated with Hp expression in brain, reverse transcription-polymerase chain reaction (RT-PCR) experiments were conducted to determine if the Hp mRNA transcript is expressed in human glioblastoma cells. Furthermore, Western blots and immunoprecipitations were performed to elucidate if Hp protein is synthesized and secreted by human glioblastoma cells. The Hp mRNA (alpha2beta) transcript (1155 bp) was detected both in U-87MG and U-138MG cells, and was positively verified by nested PCR in which a part of the beta sequence (482 bp) was targeted for amplification. Despite the presence of Hp mRNA, Hp protein was not secreted by U-87MG cells as compared to the hepatoma cell line, HepG2, where Hp protein (approximately 46 kDa) was detected in the media. The results suggest the expression of Hp protein by glioblastoma cells is possible since the Hp mRNA transcript exist, but whether or not Hp mRNA is contained in a storage pool requiring a specific signal for translation or is transiently expressed remains to be uncovered in future studies.
Sanchez
D J
DJ
Department of Biology, University of Texas at San Antonio, 6900 NW Loop 1604, San Antonio, TX 78249, USA.
Armstrong
L
L
Aguilar
R
R
Adrian
G S
GS
Haro
L
L
Martinez
A O
AO
D000544:Alzheimer Disease / Q000134:cerebrospinal fluid* / Q000235:genetics / Q000503:physiopathology; D001932:Brain Neoplasms / Q000134:cerebrospinal fluid* / Q000235:genetics / Q000503:physiopathology; D004660:Encephalitis / Q000134:cerebrospinal fluid / Q000235:genetics / Q000503:physiopathology; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D005909:Glioblastoma / Q000134:cerebrospinal fluid* / Q000235:genetics / Q000503:physiopathology; D006242:Haptoglobins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D012333:RNA, Messenger / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism*
D006242:Haptoglobins; D012333:RNA, Messenger