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CXC receptor-4 mRNA silencing abrogates CXCL12-induced migration of colorectal cancer cells.

Rubie C, Frick VO, Ghadjar P, Wagner M, Justinger C, Faust SK, Vicinus B, Gräber S, Kollmar O, Schilling MK - J Transl Med (2011)

Bottom Line: In CRC tissues CXCL12 was significantly down-regulated and CXCR4 was significantly up-regulated compared to the corresponding normal tissues.This effect was significantly abrogated by neutralizing anti-CXCR4 antibody as well as by CXCR4 siRNAs (P < 0.05).Thus, the expression and functionality of CXCR4 might be associated with the metastatic potential of CRC cells and CXCL12/CXCR4 interactions might therefore constitute a promising target for specific treatment interventions.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General -, Visceral-, Vascular - and Paediatric Surgery, University of the Saarland, 66421 Homburg/Saar, Germany. claudia.rubie@uks.eu

ABSTRACT

Background: Interactions between CXCR4 and its ligand CXCL12 have been shown to be involved in cancer progression in colorectal cancer (CRC). We performed a comparative CXCL12/CXCR4 expression analysis and assessed the effect of external CXCL12 stimulation on migration of CRC cells without and with CXCR4 inhibition.

Methods: Expression of CXCL12/CXCR4 was assessed by quantitative real-time PCR, ELISA and immunohistochemistry in resection specimens of 50 CRC patients as well as in the corresponding normal tissues and in three human CRC cell lines with different metastatic potential (Caco-2, SW480 and HT-29). Migration assays were performed after stimulation with CXCL12 and CXCR4 was inhibited by siRNA and neutralizing antibodies.

Results: In CRC tissues CXCL12 was significantly down-regulated and CXCR4 was significantly up-regulated compared to the corresponding normal tissues. In cell lines CXCR4 was predominantly expressed in SW480 and less pronounced in HT-29 cells. CXCL12 was only detectable in Caco-2 cells. CXCL12 stimulation had no impact on Caco-2 cells but significantly increased migration of CXCR4 bearing SW480 and HT-29 cells. This effect was significantly abrogated by neutralizing anti-CXCR4 antibody as well as by CXCR4 siRNAs (P < 0.05).

Conclusions: CXCR4 expression was up-regulated in CRC and CXCL12 stimulation increased migration in CXCR4 bearing cell lines. Migration was inhibited by both neutralizing CXCR4 antibodies and CXCR4 siRNAs. Thus, the expression and functionality of CXCR4 might be associated with the metastatic potential of CRC cells and CXCL12/CXCR4 interactions might therefore constitute a promising target for specific treatment interventions.

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CXCR4 protein inhibition abrogates migration of colorectal cancer cells. (A) Percent Caco-2 cell migration related to a positive control containing 20% FCS (PC) without inhibition and after inhibition with anti-CXCR4 antibody. (B) Percent HT-29 cell migration related to PC without inhibition and after inhibition with anti-CXCR4 antibody. (C) Percent SW480 cell migration related to PC without inhibition and after inhibition with anti-CXCR4 antibody.
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Figure 4: CXCR4 protein inhibition abrogates migration of colorectal cancer cells. (A) Percent Caco-2 cell migration related to a positive control containing 20% FCS (PC) without inhibition and after inhibition with anti-CXCR4 antibody. (B) Percent HT-29 cell migration related to PC without inhibition and after inhibition with anti-CXCR4 antibody. (C) Percent SW480 cell migration related to PC without inhibition and after inhibition with anti-CXCR4 antibody.

Mentions: All three cell lines were stimulated with 10, 50 and 100 ng/ml CXCL12, respectively, and incubated for 48 hours. While Caco-2 cells were not stimulated by any concentration of CXCL12 (Figure 4A), we observed a significant CXCL12 dose-independent stimulation of migration for HT-29 and SW480 cells (Figure 4B and 4C, respectively) (P < 0.05). Thus, CXCL12 was shown to be chemotactic for HT-29 and SW480 cells.


CXC receptor-4 mRNA silencing abrogates CXCL12-induced migration of colorectal cancer cells.

Rubie C, Frick VO, Ghadjar P, Wagner M, Justinger C, Faust SK, Vicinus B, Gräber S, Kollmar O, Schilling MK - J Transl Med (2011)

CXCR4 protein inhibition abrogates migration of colorectal cancer cells. (A) Percent Caco-2 cell migration related to a positive control containing 20% FCS (PC) without inhibition and after inhibition with anti-CXCR4 antibody. (B) Percent HT-29 cell migration related to PC without inhibition and after inhibition with anti-CXCR4 antibody. (C) Percent SW480 cell migration related to PC without inhibition and after inhibition with anti-CXCR4 antibody.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3049756&req=5

Figure 4: CXCR4 protein inhibition abrogates migration of colorectal cancer cells. (A) Percent Caco-2 cell migration related to a positive control containing 20% FCS (PC) without inhibition and after inhibition with anti-CXCR4 antibody. (B) Percent HT-29 cell migration related to PC without inhibition and after inhibition with anti-CXCR4 antibody. (C) Percent SW480 cell migration related to PC without inhibition and after inhibition with anti-CXCR4 antibody.
Mentions: All three cell lines were stimulated with 10, 50 and 100 ng/ml CXCL12, respectively, and incubated for 48 hours. While Caco-2 cells were not stimulated by any concentration of CXCL12 (Figure 4A), we observed a significant CXCL12 dose-independent stimulation of migration for HT-29 and SW480 cells (Figure 4B and 4C, respectively) (P < 0.05). Thus, CXCL12 was shown to be chemotactic for HT-29 and SW480 cells.

Bottom Line: In CRC tissues CXCL12 was significantly down-regulated and CXCR4 was significantly up-regulated compared to the corresponding normal tissues.This effect was significantly abrogated by neutralizing anti-CXCR4 antibody as well as by CXCR4 siRNAs (P < 0.05).Thus, the expression and functionality of CXCR4 might be associated with the metastatic potential of CRC cells and CXCL12/CXCR4 interactions might therefore constitute a promising target for specific treatment interventions.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General -, Visceral-, Vascular - and Paediatric Surgery, University of the Saarland, 66421 Homburg/Saar, Germany. claudia.rubie@uks.eu

ABSTRACT

Background: Interactions between CXCR4 and its ligand CXCL12 have been shown to be involved in cancer progression in colorectal cancer (CRC). We performed a comparative CXCL12/CXCR4 expression analysis and assessed the effect of external CXCL12 stimulation on migration of CRC cells without and with CXCR4 inhibition.

Methods: Expression of CXCL12/CXCR4 was assessed by quantitative real-time PCR, ELISA and immunohistochemistry in resection specimens of 50 CRC patients as well as in the corresponding normal tissues and in three human CRC cell lines with different metastatic potential (Caco-2, SW480 and HT-29). Migration assays were performed after stimulation with CXCL12 and CXCR4 was inhibited by siRNA and neutralizing antibodies.

Results: In CRC tissues CXCL12 was significantly down-regulated and CXCR4 was significantly up-regulated compared to the corresponding normal tissues. In cell lines CXCR4 was predominantly expressed in SW480 and less pronounced in HT-29 cells. CXCL12 was only detectable in Caco-2 cells. CXCL12 stimulation had no impact on Caco-2 cells but significantly increased migration of CXCR4 bearing SW480 and HT-29 cells. This effect was significantly abrogated by neutralizing anti-CXCR4 antibody as well as by CXCR4 siRNAs (P < 0.05).

Conclusions: CXCR4 expression was up-regulated in CRC and CXCL12 stimulation increased migration in CXCR4 bearing cell lines. Migration was inhibited by both neutralizing CXCR4 antibodies and CXCR4 siRNAs. Thus, the expression and functionality of CXCR4 might be associated with the metastatic potential of CRC cells and CXCL12/CXCR4 interactions might therefore constitute a promising target for specific treatment interventions.

Show MeSH
Related in: MedlinePlus