Transcriptional activation of TMSG-1 by complex of KLF6 and Sp1 / 中华病理学杂志
Chinese Journal of Pathology
;
(12): 542-548, 2011.
Article
in Chinese
| WPRIM
| ID: wpr-358303
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the regulatory mechanism of the transcription of tumor metastasis suppressor gene TMSG-1.</p><p><b>METHODS</b>Luciferase reporter assay and site-directed mutagenesis were used to analyze the regulatory region of TMSG-1. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were carried out to verify the interaction of KLF6 and Sp1 with the regulatory region of TMSG-1. Co-immunoprecipitation (CoIP) was performed to analyze the interaction between KLF6 and Sp1. TMSG-1 and wt-KLF6 mRNA expressions in cells with different metastatic capacities were quantitated by real-time PCR. Cell invasive capability was determined by Matrigel invasion assay.</p><p><b>RESULTS</b>A 63 bp inducible regulatory region (+59 bp - +123 bp) in exon 1 was identified by luciferase assay using reporter plasmids with a series of TMSG-1 regulatory region deletions. Mutations in KLF6/Sp1 binding sites of this region resulted in a decrease of luciferase activity, while cotransfection with KLF6 or Sp1 expressing plasmids led to a remarkable increase of luciferase activity. EMSA and ChIP demonstrated that KLF6 as well as Sp1 interacted with this region. CoIP also indicated a possible interaction between KLF6 and Sp1 proteins. In the highly metastatic cell sublines, a low level of wild type KLF6 was associated synchronously with a low TMSG-1 level. Prostate carcinoma cells overexpressing KLF6 exhibited a higher TMSG-1 level and a lower invasive capability.</p><p><b>CONCLUSIONS</b>Transcription factor complex of KLF6 and Sp1 may participate in the inducible transcriptional regulation of TMSG-1, and a decreased wild type KLF6 expression is likely associated with a low TMSG-1 level in the highly metastatic cell sublines.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Prostatic Neoplasms
/
Binding Sites
/
RNA, Messenger
/
Transfection
/
Transcriptional Activation
/
Mutagenesis, Site-Directed
/
Sp1 Transcription Factor
/
Proto-Oncogene Proteins
/
Tumor Suppressor Proteins
Type of study:
Prognostic study
Limits:
Humans
/
Male
Language:
Chinese
Journal:
Chinese Journal of Pathology
Year:
2011
Type:
Article
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