Minimal and inducible regulation of tissue factor pathway inhibitor-2 in human gliomas

Santhi D. Konduri, Francis Ali Osman, Chilukuri N. Rao, Harish Srinivas, Niranjan Yanamandra, Anastasia Tasiou, Dzung H. Dinh, William C. Olivero, Meena Gujrati, Donald C. Foster, Walter Kisiel, Gregory Kouraklis, Jasti S. Rao

Research output: Contribution to journalArticle

Abstract

Tissue factor pathway inhibitor-2 (TFPI-2), a serine protease inhibitor abundant in the extra cellular matrix, is highly expressed in non-invasive cells but undetectable levels in highly invasive human glioma cells. The mechanisms responsible for its transcriptional regulation are not well elucidated. In this study, we made several deletion constructs from a 3.6 kb genomic fragment from Hs683 cells containing the 5′-flanking region of the TFPI-2 gene, transiently transfected with these constructs into non-invasive (Hs683) and highly invasive (SNB19) human glioma cells, and assessed their expression by using a luciferase reporter gene. Three constructs showed high promoter activity (pTF5, -670 to +1; pTF6, -312 to +1; pTF2, -1511 to +1). Another construct, pTF8 (-81 to +1), showed no activity. PTF9, a variant of pTF5 in which a further 231 bp fragment (-312 to -81) was deleted, from the [-670 to +1] pTF5 region, also showed no promoter activity. Hence, (-312 to -81) this region is essential for the transcription of TFPI-2 in glioma cells. Sequencing of this promoter region revealed that it has a high G+C content, contains potential SP1 and AP1 binding motifs, and lacks canonical TATA and CAAT boxes immediately upstream of the major transcriptional initiation site, although CAAT boxes were found about -3000 bp upstream of the transcription start site. We also found a strong repressor in the region between -927 to -1181, upstream of the major transcriptional initiation site, followed by positive elements or enhancers between -1511 to -1181. These positive elements masked the silencer effect. Finally TFPI-2 was induced in Hs683 cells transfected with the pTF6 construct (-312 to +1) and stimulated with phorbol-12-myristate-13-acetate (PMA). We conclude that the -312 to + 1 region is critical for the minimal and inducible regulation of TFPI-2 in non-invasive (Hs683) and highly invasive (SNB19) human glioma cell lines.

Original languageEnglish (US)
Pages (from-to)921-928
Number of pages8
JournalOncogene
Volume21
Issue number6
DOIs
StatePublished - Jan 31 2002

Fingerprint

Glioma
Transcriptional Silencer Elements
TATA Box
Serine Proteinase Inhibitors
5' Flanking Region
Transcription Initiation Site
Base Composition
Luciferases
Reporter Genes
Genetic Promoter Regions
tissue-factor-pathway inhibitor 2
Acetates
Transcription Factors
Cell Line
Genes

Keywords

  • Gliomas
  • Promoter
  • TFPI-2
  • Transcriptional regulation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

Cite this

Konduri, S. D., Osman, F. A., Rao, C. N., Srinivas, H., Yanamandra, N., Tasiou, A., ... Rao, J. S. (2002). Minimal and inducible regulation of tissue factor pathway inhibitor-2 in human gliomas. Oncogene, 21(6), 921-928. https://doi.org/10.1038/sj/onc/1204983

Minimal and inducible regulation of tissue factor pathway inhibitor-2 in human gliomas. / Konduri, Santhi D.; Osman, Francis Ali; Rao, Chilukuri N.; Srinivas, Harish; Yanamandra, Niranjan; Tasiou, Anastasia; Dinh, Dzung H.; Olivero, William C.; Gujrati, Meena; Foster, Donald C.; Kisiel, Walter; Kouraklis, Gregory; Rao, Jasti S.

In: Oncogene, Vol. 21, No. 6, 31.01.2002, p. 921-928.

Research output: Contribution to journalArticle

Konduri, SD, Osman, FA, Rao, CN, Srinivas, H, Yanamandra, N, Tasiou, A, Dinh, DH, Olivero, WC, Gujrati, M, Foster, DC, Kisiel, W, Kouraklis, G & Rao, JS 2002, 'Minimal and inducible regulation of tissue factor pathway inhibitor-2 in human gliomas', Oncogene, vol. 21, no. 6, pp. 921-928. https://doi.org/10.1038/sj/onc/1204983
Konduri SD, Osman FA, Rao CN, Srinivas H, Yanamandra N, Tasiou A et al. Minimal and inducible regulation of tissue factor pathway inhibitor-2 in human gliomas. Oncogene. 2002 Jan 31;21(6):921-928. https://doi.org/10.1038/sj/onc/1204983
Konduri, Santhi D. ; Osman, Francis Ali ; Rao, Chilukuri N. ; Srinivas, Harish ; Yanamandra, Niranjan ; Tasiou, Anastasia ; Dinh, Dzung H. ; Olivero, William C. ; Gujrati, Meena ; Foster, Donald C. ; Kisiel, Walter ; Kouraklis, Gregory ; Rao, Jasti S. / Minimal and inducible regulation of tissue factor pathway inhibitor-2 in human gliomas. In: Oncogene. 2002 ; Vol. 21, No. 6. pp. 921-928.
@article{79ee30ae09994defaf5b092a86d880d2,
title = "Minimal and inducible regulation of tissue factor pathway inhibitor-2 in human gliomas",
abstract = "Tissue factor pathway inhibitor-2 (TFPI-2), a serine protease inhibitor abundant in the extra cellular matrix, is highly expressed in non-invasive cells but undetectable levels in highly invasive human glioma cells. The mechanisms responsible for its transcriptional regulation are not well elucidated. In this study, we made several deletion constructs from a 3.6 kb genomic fragment from Hs683 cells containing the 5′-flanking region of the TFPI-2 gene, transiently transfected with these constructs into non-invasive (Hs683) and highly invasive (SNB19) human glioma cells, and assessed their expression by using a luciferase reporter gene. Three constructs showed high promoter activity (pTF5, -670 to +1; pTF6, -312 to +1; pTF2, -1511 to +1). Another construct, pTF8 (-81 to +1), showed no activity. PTF9, a variant of pTF5 in which a further 231 bp fragment (-312 to -81) was deleted, from the [-670 to +1] pTF5 region, also showed no promoter activity. Hence, (-312 to -81) this region is essential for the transcription of TFPI-2 in glioma cells. Sequencing of this promoter region revealed that it has a high G+C content, contains potential SP1 and AP1 binding motifs, and lacks canonical TATA and CAAT boxes immediately upstream of the major transcriptional initiation site, although CAAT boxes were found about -3000 bp upstream of the transcription start site. We also found a strong repressor in the region between -927 to -1181, upstream of the major transcriptional initiation site, followed by positive elements or enhancers between -1511 to -1181. These positive elements masked the silencer effect. Finally TFPI-2 was induced in Hs683 cells transfected with the pTF6 construct (-312 to +1) and stimulated with phorbol-12-myristate-13-acetate (PMA). We conclude that the -312 to + 1 region is critical for the minimal and inducible regulation of TFPI-2 in non-invasive (Hs683) and highly invasive (SNB19) human glioma cell lines.",
keywords = "Gliomas, Promoter, TFPI-2, Transcriptional regulation",
author = "Konduri, {Santhi D.} and Osman, {Francis Ali} and Rao, {Chilukuri N.} and Harish Srinivas and Niranjan Yanamandra and Anastasia Tasiou and Dinh, {Dzung H.} and Olivero, {William C.} and Meena Gujrati and Foster, {Donald C.} and Walter Kisiel and Gregory Kouraklis and Rao, {Jasti S.}",
year = "2002",
month = "1",
day = "31",
doi = "10.1038/sj/onc/1204983",
language = "English (US)",
volume = "21",
pages = "921--928",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "6",

}

TY - JOUR

T1 - Minimal and inducible regulation of tissue factor pathway inhibitor-2 in human gliomas

AU - Konduri, Santhi D.

AU - Osman, Francis Ali

AU - Rao, Chilukuri N.

AU - Srinivas, Harish

AU - Yanamandra, Niranjan

AU - Tasiou, Anastasia

AU - Dinh, Dzung H.

AU - Olivero, William C.

AU - Gujrati, Meena

AU - Foster, Donald C.

AU - Kisiel, Walter

AU - Kouraklis, Gregory

AU - Rao, Jasti S.

PY - 2002/1/31

Y1 - 2002/1/31

N2 - Tissue factor pathway inhibitor-2 (TFPI-2), a serine protease inhibitor abundant in the extra cellular matrix, is highly expressed in non-invasive cells but undetectable levels in highly invasive human glioma cells. The mechanisms responsible for its transcriptional regulation are not well elucidated. In this study, we made several deletion constructs from a 3.6 kb genomic fragment from Hs683 cells containing the 5′-flanking region of the TFPI-2 gene, transiently transfected with these constructs into non-invasive (Hs683) and highly invasive (SNB19) human glioma cells, and assessed their expression by using a luciferase reporter gene. Three constructs showed high promoter activity (pTF5, -670 to +1; pTF6, -312 to +1; pTF2, -1511 to +1). Another construct, pTF8 (-81 to +1), showed no activity. PTF9, a variant of pTF5 in which a further 231 bp fragment (-312 to -81) was deleted, from the [-670 to +1] pTF5 region, also showed no promoter activity. Hence, (-312 to -81) this region is essential for the transcription of TFPI-2 in glioma cells. Sequencing of this promoter region revealed that it has a high G+C content, contains potential SP1 and AP1 binding motifs, and lacks canonical TATA and CAAT boxes immediately upstream of the major transcriptional initiation site, although CAAT boxes were found about -3000 bp upstream of the transcription start site. We also found a strong repressor in the region between -927 to -1181, upstream of the major transcriptional initiation site, followed by positive elements or enhancers between -1511 to -1181. These positive elements masked the silencer effect. Finally TFPI-2 was induced in Hs683 cells transfected with the pTF6 construct (-312 to +1) and stimulated with phorbol-12-myristate-13-acetate (PMA). We conclude that the -312 to + 1 region is critical for the minimal and inducible regulation of TFPI-2 in non-invasive (Hs683) and highly invasive (SNB19) human glioma cell lines.

AB - Tissue factor pathway inhibitor-2 (TFPI-2), a serine protease inhibitor abundant in the extra cellular matrix, is highly expressed in non-invasive cells but undetectable levels in highly invasive human glioma cells. The mechanisms responsible for its transcriptional regulation are not well elucidated. In this study, we made several deletion constructs from a 3.6 kb genomic fragment from Hs683 cells containing the 5′-flanking region of the TFPI-2 gene, transiently transfected with these constructs into non-invasive (Hs683) and highly invasive (SNB19) human glioma cells, and assessed their expression by using a luciferase reporter gene. Three constructs showed high promoter activity (pTF5, -670 to +1; pTF6, -312 to +1; pTF2, -1511 to +1). Another construct, pTF8 (-81 to +1), showed no activity. PTF9, a variant of pTF5 in which a further 231 bp fragment (-312 to -81) was deleted, from the [-670 to +1] pTF5 region, also showed no promoter activity. Hence, (-312 to -81) this region is essential for the transcription of TFPI-2 in glioma cells. Sequencing of this promoter region revealed that it has a high G+C content, contains potential SP1 and AP1 binding motifs, and lacks canonical TATA and CAAT boxes immediately upstream of the major transcriptional initiation site, although CAAT boxes were found about -3000 bp upstream of the transcription start site. We also found a strong repressor in the region between -927 to -1181, upstream of the major transcriptional initiation site, followed by positive elements or enhancers between -1511 to -1181. These positive elements masked the silencer effect. Finally TFPI-2 was induced in Hs683 cells transfected with the pTF6 construct (-312 to +1) and stimulated with phorbol-12-myristate-13-acetate (PMA). We conclude that the -312 to + 1 region is critical for the minimal and inducible regulation of TFPI-2 in non-invasive (Hs683) and highly invasive (SNB19) human glioma cell lines.

KW - Gliomas

KW - Promoter

KW - TFPI-2

KW - Transcriptional regulation

UR - http://www.scopus.com/inward/record.url?scp=18244374006&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=18244374006&partnerID=8YFLogxK

U2 - 10.1038/sj/onc/1204983

DO - 10.1038/sj/onc/1204983

M3 - Article

VL - 21

SP - 921

EP - 928

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 6

ER -