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Regulation of MCP-1 chemokine transcription by p53.

Hacke K, Rincon-Orozco B, Buchwalter G, Siehler SY, Wasylyk B, Wiesmüller L, Rösl F - Mol. Cancer (2010)

Bottom Line: In both cases, non-functional p53 leads to diminished MCP-1 transcription upon TNF-alpha treatment.In addition, siRNA directed against p53 decreased MCP-1 transcription after TNF-alpha addition, directly confirming a crosstalk between p53 and MCP-1.These data support the concept that p53 inactivation during carcinogenesis also affects immune surveillance by interfering with chemokine expression and in turn communication with cells of the immunological compartment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Deutsches Krebsforschungszentrum, Forschungsschwerpunkt Infektion und Krebs, Abteilung Virale Transformationsmechanismen, Heidelberg, Germany.

ABSTRACT

Background: Our previous studies showed that the expression of the monocyte-chemoattractant protein (MCP)-1, a chemokine, which triggers the infiltration and activation of cells of the monocyte-macrophage lineage, is abrogated in human papillomavirus (HPV)-positive premalignant and malignant cells. In silico analysis of the MCP-1 upstream region proposed a putative p53 binding side about 2.5 kb upstream of the transcriptional start. The aim of this study is to monitor a physiological role of p53 in this process.

Results: The proposed p53 binding side could be confirmed in vitro by electrophoretic-mobility-shift assays and in vivo by chromatin immunoprecipitation. Moreover, the availability of p53 is apparently important for chemokine regulation, since TNF-alpha can induce MCP-1 only in human keratinocytes expressing the viral oncoprotein E7, but not in HPV16 E6 positive cells, where p53 becomes degraded. A general physiological role of p53 in MCP-1 regulation was further substantiated in HPV-negative cells harboring a temperature-sensitive mutant of p53 and in Li-Fraumeni cells, carrying a germ-line mutation of p53. In both cases, non-functional p53 leads to diminished MCP-1 transcription upon TNF-alpha treatment. In addition, siRNA directed against p53 decreased MCP-1 transcription after TNF-alpha addition, directly confirming a crosstalk between p53 and MCP-1.

Conclusion: These data support the concept that p53 inactivation during carcinogenesis also affects immune surveillance by interfering with chemokine expression and in turn communication with cells of the immunological compartment.

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Related in: MedlinePlus

MCP-1 induction by TNF-α after reconstitution of wild-type p53 activities. 4Bv (A) and BT2E (B) cells were cultivated at 37°C or shifted to 32°C overnight. Stimulation was done with TNF-α for 4 h or 6 h. Total cellular RNA was separated in a 1% agarose gel and transferred to a Gene screen Plus membrane. The filter was subsequently hybridized with a p21, MCP-1 and c-myc cDNA probe. The positions of the 28S and 18S ribosomal RNA are indicated.
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Figure 2: MCP-1 induction by TNF-α after reconstitution of wild-type p53 activities. 4Bv (A) and BT2E (B) cells were cultivated at 37°C or shifted to 32°C overnight. Stimulation was done with TNF-α for 4 h or 6 h. Total cellular RNA was separated in a 1% agarose gel and transferred to a Gene screen Plus membrane. The filter was subsequently hybridized with a p21, MCP-1 and c-myc cDNA probe. The positions of the 28S and 18S ribosomal RNA are indicated.

Mentions: As depicted in Fig. 2A, transcription of p21 mRNA was strongly induced in 4Bv cells after the shift to 32°C, when compared to the cells left at 37°C, independently of whether TNF-α was added or not. In BT2E control cells, however, no p21 induction was observed (Fig. 2B). This clearly indicates that p53 in 4Bv cells was functionally reconstituted at 32°C.


Regulation of MCP-1 chemokine transcription by p53.

Hacke K, Rincon-Orozco B, Buchwalter G, Siehler SY, Wasylyk B, Wiesmüller L, Rösl F - Mol. Cancer (2010)

MCP-1 induction by TNF-α after reconstitution of wild-type p53 activities. 4Bv (A) and BT2E (B) cells were cultivated at 37°C or shifted to 32°C overnight. Stimulation was done with TNF-α for 4 h or 6 h. Total cellular RNA was separated in a 1% agarose gel and transferred to a Gene screen Plus membrane. The filter was subsequently hybridized with a p21, MCP-1 and c-myc cDNA probe. The positions of the 28S and 18S ribosomal RNA are indicated.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2864217&req=5

Figure 2: MCP-1 induction by TNF-α after reconstitution of wild-type p53 activities. 4Bv (A) and BT2E (B) cells were cultivated at 37°C or shifted to 32°C overnight. Stimulation was done with TNF-α for 4 h or 6 h. Total cellular RNA was separated in a 1% agarose gel and transferred to a Gene screen Plus membrane. The filter was subsequently hybridized with a p21, MCP-1 and c-myc cDNA probe. The positions of the 28S and 18S ribosomal RNA are indicated.
Mentions: As depicted in Fig. 2A, transcription of p21 mRNA was strongly induced in 4Bv cells after the shift to 32°C, when compared to the cells left at 37°C, independently of whether TNF-α was added or not. In BT2E control cells, however, no p21 induction was observed (Fig. 2B). This clearly indicates that p53 in 4Bv cells was functionally reconstituted at 32°C.

Bottom Line: In both cases, non-functional p53 leads to diminished MCP-1 transcription upon TNF-alpha treatment.In addition, siRNA directed against p53 decreased MCP-1 transcription after TNF-alpha addition, directly confirming a crosstalk between p53 and MCP-1.These data support the concept that p53 inactivation during carcinogenesis also affects immune surveillance by interfering with chemokine expression and in turn communication with cells of the immunological compartment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Deutsches Krebsforschungszentrum, Forschungsschwerpunkt Infektion und Krebs, Abteilung Virale Transformationsmechanismen, Heidelberg, Germany.

ABSTRACT

Background: Our previous studies showed that the expression of the monocyte-chemoattractant protein (MCP)-1, a chemokine, which triggers the infiltration and activation of cells of the monocyte-macrophage lineage, is abrogated in human papillomavirus (HPV)-positive premalignant and malignant cells. In silico analysis of the MCP-1 upstream region proposed a putative p53 binding side about 2.5 kb upstream of the transcriptional start. The aim of this study is to monitor a physiological role of p53 in this process.

Results: The proposed p53 binding side could be confirmed in vitro by electrophoretic-mobility-shift assays and in vivo by chromatin immunoprecipitation. Moreover, the availability of p53 is apparently important for chemokine regulation, since TNF-alpha can induce MCP-1 only in human keratinocytes expressing the viral oncoprotein E7, but not in HPV16 E6 positive cells, where p53 becomes degraded. A general physiological role of p53 in MCP-1 regulation was further substantiated in HPV-negative cells harboring a temperature-sensitive mutant of p53 and in Li-Fraumeni cells, carrying a germ-line mutation of p53. In both cases, non-functional p53 leads to diminished MCP-1 transcription upon TNF-alpha treatment. In addition, siRNA directed against p53 decreased MCP-1 transcription after TNF-alpha addition, directly confirming a crosstalk between p53 and MCP-1.

Conclusion: These data support the concept that p53 inactivation during carcinogenesis also affects immune surveillance by interfering with chemokine expression and in turn communication with cells of the immunological compartment.

Show MeSH
Related in: MedlinePlus