Molecular mechanisms involved in transcriptional activation of the human Sia-alpha2,3-Gal-beta1,4-GlcNAc-R:alpha2,8-sialyltransferase (hST8Sia III) gene induced by KCl in human glioblastoma cells.
In the present study, we have shown that KCl known as an inducer for differentiation of neuronal cells increases the human Sia-alpha2,3-Gal-beta1,4-GlcNAc-R:alpha2,8-sialyltransferase (hST8Sia III) gene transcription via phosphoinositide 3 kinase (PI-3K) in glioblastoma U-87MG cells. The induction of hST8Sia III by KCl is regulated at the transcriptional level in a dose- and time-dependent manner as evidenced by reverse transcription-polymerase chain reaction (RT-PCR). To elucidate the mechanism underlying the regulation of hST8Sia III gene expression in U-87MG cells induced by KCl, we characterized the promoter region of the hST8Sia III gene. Functional analysis of the 5'-flanking region of the hST8Sia III gene by the transient expression method showed that the -1194 to -816 region functions as the KCl-inducible promoter in U-87MG cells. Furthermore, as evidenced by Western blot analysis and RT-PCR, KCl-induced expression of hST8Sia III gene was dependent on the PI-3K signal transduction pathway during the neuronal differentiation of U-87 cells, as an increase in beta-tubulin III known as a neuronal differentiation marker was observed. In KCl-depolarization on U-87 cells, the PI-3K-dependent promoter activation at the -1194 to -816 region up-regulated expression of hST8Sia III gene. These results suggest that the expression of hST8Sia III gene via the PI-3K signaling pathway is enhanced during KCl-induced differentiation of U-87 cells by increasing expression of beta-tubulin III.
Kim
Seok-Jo
SJ
Department of Biotechnology, Dong-A University, Busan 604-714, Republic of Korea.
Chung
Tae-Wook
TW
Jin
Un-Ho
UH
Suh
Seok-Jong
SJ
Lee
Young-Choon
YC
Kim
Cheorl-Ho
CH
D024506:5' Flanking Region; D001483:Base Sequence; D002454:Cell Differentiation / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D005909:Glioblastoma; D006801:Humans; D008969:Molecular Sequence Data; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011189:Potassium Chloride / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D012799:Sialyltransferases / Q000235:genetics*; D024363:Transcription Initiation Site; D014158:Transcription, Genetic / Q000187:drug effects; D015533:Transcriptional Activation; D014404:Tubulin / Q000378:metabolism
C498585:TUBB3 protein, human; D014404:Tubulin; D011189:Potassium Chloride; C094133:Sia(alpha2,3)Gal(beta1,4)GlcNAc alpha-2,8-sialyltransferase; D012799:Sialyltransferases; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt