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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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CXCL12 protein expression in different human organs related to the liver as determined by ELISA. CXCL12 protein concentrations (pg/ml per mg total protein) were measured in total cell lysates of normal liver, pancreas, stomach, colon/rectum and esophagus tissues of 10 patients, respectively (n = 10). Protein data are expressed as mean +/- SEM, *P < 0.05.
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Figure 2: CXCL12 protein expression in different human organs related to the liver as determined by ELISA. CXCL12 protein concentrations (pg/ml per mg total protein) were measured in total cell lysates of normal liver, pancreas, stomach, colon/rectum and esophagus tissues of 10 patients, respectively (n = 10). Protein data are expressed as mean +/- SEM, *P < 0.05.

Mentions: CXCL12 mRNA and protein expression as determined by RT-PCR and ELISA was investigated in different human organs such as pancreas, stomach, colon/rectum and esophagus related to the liver. On the protein level CXCL12 was found to exhibit peak levels of expression in the liver compared to stomach, esophagus, pancreas, colon and rectum (Figure 2). However, on the mRNA level CXCL12 did not show a markedly higher expression in the liver in comparison to other gastrointestinal organs or glands corresponding to previous results [27].


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)

CXCL12 protein expression in different human organs related to the liver as determined by ELISA. CXCL12 protein concentrations (pg/ml per mg total protein) were measured in total cell lysates of normal liver, pancreas, stomach, colon/rectum and esophagus tissues of 10 patients, respectively (n = 10). Protein data are expressed as mean +/- SEM, *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: CXCL12 protein expression in different human organs related to the liver as determined by ELISA. CXCL12 protein concentrations (pg/ml per mg total protein) were measured in total cell lysates of normal liver, pancreas, stomach, colon/rectum and esophagus tissues of 10 patients, respectively (n = 10). Protein data are expressed as mean +/- SEM, *P < 0.05.
Mentions: CXCL12 mRNA and protein expression as determined by RT-PCR and ELISA was investigated in different human organs such as pancreas, stomach, colon/rectum and esophagus related to the liver. On the protein level CXCL12 was found to exhibit peak levels of expression in the liver compared to stomach, esophagus, pancreas, colon and rectum (Figure 2). However, on the mRNA level CXCL12 did not show a markedly higher expression in the liver in comparison to other gastrointestinal organs or glands corresponding to previous results [27].

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