Limits...
CXCR4 expression affects overall survival of HCC patients whereas CXCR7 expression does not.

Neve Polimeno M, Ierano C, D'Alterio C, Simona Losito N, Napolitano M, Portella L, Scognamiglio G, Tatangelo F, Maria Trotta A, Curley S, Costantini S, Liuzzi R, Izzo F, Scala S - Cell. Mol. Immunol. (2014)

Bottom Line: Interestingly, the common CXCR4-CXCR7 ligand CXCL12 was expressed at significantly lower levels in tumor tissues compared to adjacent normal liver (P=0.032).In conclusion, CXCR4 affects the prognosis of HCC patients but CXCR7 does not.Therefore, the CXCR4-CXCL12-CXCR7 axis plays a role in the interaction of HCC with the surrounding normal tissue and represents a suitable therapeutic target.

View Article: PubMed Central - PubMed

Affiliation: Oncological Immunology, Istituto Nazionale per lo Studio e la Cura dei Tumori, "Fondazione Giovanni Pascale"-IRCCS-ITALIA, Naples, Italy.

ABSTRACT
Hepatocellular carcinoma (HCC) is a heterogeneous disease with a poor prognosis and limited markers for predicting patient survival. Because chemokines and chemokine receptors play numerous and integral roles in HCC disease progression, the CXCR4-CXCL12-CXCR7 axis was studied in HCC patients. CXCR4 and CXCR7 expression was analyzed by immunohistochemistry in 86 HCC patients (training cohort) and validated in 42 unrelated HCC patients (validation cohort). CXCR4 levels were low in 22.1% of patients, intermediate in 30.2%, and high in 47.7%, whereas CXCR7 levels were low in 9.3% of patients, intermediate in 44.2% and high in 46.5% of the patients in the training cohort. When correlated to patient outcome, only CXCR4 affected overall survival (P=0.03). CXCR4-CXCL12-CXCR7 mRNA levels were examined in 33/86 patients. Interestingly, the common CXCR4-CXCR7 ligand CXCL12 was expressed at significantly lower levels in tumor tissues compared to adjacent normal liver (P=0.032). The expression and function of CXCR4 and CXCR7 was also analyzed in several human HCC cell lines. CXCR4 was expressed in Huh7, Hep3B, SNU398, SNU449 and SNU475 cells, whereas CXCR7 was expressed in HepG2, Huh7, SNU449 and SNU475 cells. Huh7, SNU449 and SNU475 cells migrated toward CXCL12, and this migration was inhibited by AMD3100/anti-CXCR4 and by CCX771/anti-CXCR7. Moreover, SNU449 and Huh7 cells exhibited matrix invasion in the presence of CXCL12 and CXCL11, a ligand exclusive to CXCR7. In conclusion, CXCR4 affects the prognosis of HCC patients but CXCR7 does not. Therefore, the CXCR4-CXCL12-CXCR7 axis plays a role in the interaction of HCC with the surrounding normal tissue and represents a suitable therapeutic target.

No MeSH data available.


Related in: MedlinePlus

CXCL12 induced migration and invasion in HCC cell lines. (a) Migration of HepG2, Hep3B, Huh7, SNU398, SNU449 and SNU475 cells. The cells were incubated in the presence of Peptide R (10 mM), AMD3100 (5 µM) or CCX771 (500 nM). Cells migrated toward CXCL12 (100 ng/ml) or CXCL11 (100 ng/ml) for 18 h. (b) CXCL12-dependent cell migration was examined in the human HCC cell lines HUH7 and SNU449 in the presence of Peptide R (5 µM), anti-CXCR4 (12G5; 10 µg/ml) or anti-CXCR7(10 µg/ml). The cells migrated toward CXCL12 (100 ng/ml) or CXCL11 (100 ng/ml) for 18 h. (c) CXCL12/CXCL11-dependent cell invasion was examined in the human HCC cell lines Huh7 and SNU449 in the presence of Peptide R (5 µM), anti-CXCR4 (12G5; 10 µg/ml) or anti-CXCR7(10 µg/ml) for 18 h. The cells were counted in 10 different consecutive high-power fields (magnification: ×200). The results represent at least three independent experiments and are expressed relative to the values for BSA alone. Statistical significance was calculated using Student's t-test. *P<0.05, **P<0.001, ***P<0.001 for BSA and the relative inhibitor. HCC, hepatocellular carcinoma.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4496532&req=5

fig5: CXCL12 induced migration and invasion in HCC cell lines. (a) Migration of HepG2, Hep3B, Huh7, SNU398, SNU449 and SNU475 cells. The cells were incubated in the presence of Peptide R (10 mM), AMD3100 (5 µM) or CCX771 (500 nM). Cells migrated toward CXCL12 (100 ng/ml) or CXCL11 (100 ng/ml) for 18 h. (b) CXCL12-dependent cell migration was examined in the human HCC cell lines HUH7 and SNU449 in the presence of Peptide R (5 µM), anti-CXCR4 (12G5; 10 µg/ml) or anti-CXCR7(10 µg/ml). The cells migrated toward CXCL12 (100 ng/ml) or CXCL11 (100 ng/ml) for 18 h. (c) CXCL12/CXCL11-dependent cell invasion was examined in the human HCC cell lines Huh7 and SNU449 in the presence of Peptide R (5 µM), anti-CXCR4 (12G5; 10 µg/ml) or anti-CXCR7(10 µg/ml) for 18 h. The cells were counted in 10 different consecutive high-power fields (magnification: ×200). The results represent at least three independent experiments and are expressed relative to the values for BSA alone. Statistical significance was calculated using Student's t-test. *P<0.05, **P<0.001, ***P<0.001 for BSA and the relative inhibitor. HCC, hepatocellular carcinoma.

Mentions: To further investigate the function of this chemokine axis in HCC, a panel of human HCC cell lines (HepG2, Huh7, Hep3B, SNU398, SNU449 and SNU475) were characterized for CXCR4 and CXCR7 expression and function. Figure 4a shows that CXCR4 protein was almost undetectable in HepG2 cells, although it was clearly expressed in Huh7, Hep3B, SNU398, SNU449 and SNU475 cells with an RNA level comparable to the protein level. CXCR7, evaluated using immunocytochemistry, showed that HepG2, Huh7, SNU449 and SNU475 cell lines overexpressed CXCR7 mainly in the cytosol (Figure 4b). CXCL12 significantly induced migration of Huh7, SNU449 and SNU475 cells, whereas migration was significantly inhibited by AMD3100 (CXCR4 antagonist) and by CCX771 (CXCR7 antagonist) in Huh7, SNU449 and SNU475 cells, suggesting a functional role for the CXCR4 and CXCR7 receptors (Figure 5a). CXCL12-dependent migration was not detectable in HepG2, Hep3B or SNU398 cells (data not shown). To further dissect the role of CXCR4 and CXCR7 receptors, the migration of Huh7 and SNU449 cells in response to CXCL12 or CXCL11, the exclusive CXCR7 ligand, was evaluated in the presence of anti-CXCR4 (12G5) (10 µg/ml), anti-CXCR7 (11G8) (10 µg/ml) or Peptide R (10 µM), a recently developed CXCR4 antagonist.28 As shown in Figure 5b, the cells migrated toward CXCL12 and CXCL11. The migration toward CXCL12 was inhibited by anti-CXCR4 and Peptide R, whereas CXCL11-induced migration was inhibited by anti-CXCR7, demonstrating that both chemokine receptors affected migration in the evaluated HCC cell lines. To further elucidate the role of CXCR4/CXCR7 in HCC cancer progression, CXCL12/CXCL11-induced cell invasion experiments were performed in Huh7 and SNU449 cells. As shown in Figure 5c, CXCL12 and CXCL11 significantly stimulated Huh7 and SNU449 invasion, which was impaired by anti-CXCR4 (12G5) and Peptide R and by anti-CXCR7 (11G8). These results suggest that CXCL12 modulates cell invasion through CXCR4/CXCR7 receptors.


CXCR4 expression affects overall survival of HCC patients whereas CXCR7 expression does not.

Neve Polimeno M, Ierano C, D'Alterio C, Simona Losito N, Napolitano M, Portella L, Scognamiglio G, Tatangelo F, Maria Trotta A, Curley S, Costantini S, Liuzzi R, Izzo F, Scala S - Cell. Mol. Immunol. (2014)

CXCL12 induced migration and invasion in HCC cell lines. (a) Migration of HepG2, Hep3B, Huh7, SNU398, SNU449 and SNU475 cells. The cells were incubated in the presence of Peptide R (10 mM), AMD3100 (5 µM) or CCX771 (500 nM). Cells migrated toward CXCL12 (100 ng/ml) or CXCL11 (100 ng/ml) for 18 h. (b) CXCL12-dependent cell migration was examined in the human HCC cell lines HUH7 and SNU449 in the presence of Peptide R (5 µM), anti-CXCR4 (12G5; 10 µg/ml) or anti-CXCR7(10 µg/ml). The cells migrated toward CXCL12 (100 ng/ml) or CXCL11 (100 ng/ml) for 18 h. (c) CXCL12/CXCL11-dependent cell invasion was examined in the human HCC cell lines Huh7 and SNU449 in the presence of Peptide R (5 µM), anti-CXCR4 (12G5; 10 µg/ml) or anti-CXCR7(10 µg/ml) for 18 h. The cells were counted in 10 different consecutive high-power fields (magnification: ×200). The results represent at least three independent experiments and are expressed relative to the values for BSA alone. Statistical significance was calculated using Student's t-test. *P<0.05, **P<0.001, ***P<0.001 for BSA and the relative inhibitor. HCC, hepatocellular carcinoma.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496532&req=5

fig5: CXCL12 induced migration and invasion in HCC cell lines. (a) Migration of HepG2, Hep3B, Huh7, SNU398, SNU449 and SNU475 cells. The cells were incubated in the presence of Peptide R (10 mM), AMD3100 (5 µM) or CCX771 (500 nM). Cells migrated toward CXCL12 (100 ng/ml) or CXCL11 (100 ng/ml) for 18 h. (b) CXCL12-dependent cell migration was examined in the human HCC cell lines HUH7 and SNU449 in the presence of Peptide R (5 µM), anti-CXCR4 (12G5; 10 µg/ml) or anti-CXCR7(10 µg/ml). The cells migrated toward CXCL12 (100 ng/ml) or CXCL11 (100 ng/ml) for 18 h. (c) CXCL12/CXCL11-dependent cell invasion was examined in the human HCC cell lines Huh7 and SNU449 in the presence of Peptide R (5 µM), anti-CXCR4 (12G5; 10 µg/ml) or anti-CXCR7(10 µg/ml) for 18 h. The cells were counted in 10 different consecutive high-power fields (magnification: ×200). The results represent at least three independent experiments and are expressed relative to the values for BSA alone. Statistical significance was calculated using Student's t-test. *P<0.05, **P<0.001, ***P<0.001 for BSA and the relative inhibitor. HCC, hepatocellular carcinoma.
Mentions: To further investigate the function of this chemokine axis in HCC, a panel of human HCC cell lines (HepG2, Huh7, Hep3B, SNU398, SNU449 and SNU475) were characterized for CXCR4 and CXCR7 expression and function. Figure 4a shows that CXCR4 protein was almost undetectable in HepG2 cells, although it was clearly expressed in Huh7, Hep3B, SNU398, SNU449 and SNU475 cells with an RNA level comparable to the protein level. CXCR7, evaluated using immunocytochemistry, showed that HepG2, Huh7, SNU449 and SNU475 cell lines overexpressed CXCR7 mainly in the cytosol (Figure 4b). CXCL12 significantly induced migration of Huh7, SNU449 and SNU475 cells, whereas migration was significantly inhibited by AMD3100 (CXCR4 antagonist) and by CCX771 (CXCR7 antagonist) in Huh7, SNU449 and SNU475 cells, suggesting a functional role for the CXCR4 and CXCR7 receptors (Figure 5a). CXCL12-dependent migration was not detectable in HepG2, Hep3B or SNU398 cells (data not shown). To further dissect the role of CXCR4 and CXCR7 receptors, the migration of Huh7 and SNU449 cells in response to CXCL12 or CXCL11, the exclusive CXCR7 ligand, was evaluated in the presence of anti-CXCR4 (12G5) (10 µg/ml), anti-CXCR7 (11G8) (10 µg/ml) or Peptide R (10 µM), a recently developed CXCR4 antagonist.28 As shown in Figure 5b, the cells migrated toward CXCL12 and CXCL11. The migration toward CXCL12 was inhibited by anti-CXCR4 and Peptide R, whereas CXCL11-induced migration was inhibited by anti-CXCR7, demonstrating that both chemokine receptors affected migration in the evaluated HCC cell lines. To further elucidate the role of CXCR4/CXCR7 in HCC cancer progression, CXCL12/CXCL11-induced cell invasion experiments were performed in Huh7 and SNU449 cells. As shown in Figure 5c, CXCL12 and CXCL11 significantly stimulated Huh7 and SNU449 invasion, which was impaired by anti-CXCR4 (12G5) and Peptide R and by anti-CXCR7 (11G8). These results suggest that CXCL12 modulates cell invasion through CXCR4/CXCR7 receptors.

Bottom Line: Interestingly, the common CXCR4-CXCR7 ligand CXCL12 was expressed at significantly lower levels in tumor tissues compared to adjacent normal liver (P=0.032).In conclusion, CXCR4 affects the prognosis of HCC patients but CXCR7 does not.Therefore, the CXCR4-CXCL12-CXCR7 axis plays a role in the interaction of HCC with the surrounding normal tissue and represents a suitable therapeutic target.

View Article: PubMed Central - PubMed

Affiliation: Oncological Immunology, Istituto Nazionale per lo Studio e la Cura dei Tumori, "Fondazione Giovanni Pascale"-IRCCS-ITALIA, Naples, Italy.

ABSTRACT
Hepatocellular carcinoma (HCC) is a heterogeneous disease with a poor prognosis and limited markers for predicting patient survival. Because chemokines and chemokine receptors play numerous and integral roles in HCC disease progression, the CXCR4-CXCL12-CXCR7 axis was studied in HCC patients. CXCR4 and CXCR7 expression was analyzed by immunohistochemistry in 86 HCC patients (training cohort) and validated in 42 unrelated HCC patients (validation cohort). CXCR4 levels were low in 22.1% of patients, intermediate in 30.2%, and high in 47.7%, whereas CXCR7 levels were low in 9.3% of patients, intermediate in 44.2% and high in 46.5% of the patients in the training cohort. When correlated to patient outcome, only CXCR4 affected overall survival (P=0.03). CXCR4-CXCL12-CXCR7 mRNA levels were examined in 33/86 patients. Interestingly, the common CXCR4-CXCR7 ligand CXCL12 was expressed at significantly lower levels in tumor tissues compared to adjacent normal liver (P=0.032). The expression and function of CXCR4 and CXCR7 was also analyzed in several human HCC cell lines. CXCR4 was expressed in Huh7, Hep3B, SNU398, SNU449 and SNU475 cells, whereas CXCR7 was expressed in HepG2, Huh7, SNU449 and SNU475 cells. Huh7, SNU449 and SNU475 cells migrated toward CXCL12, and this migration was inhibited by AMD3100/anti-CXCR4 and by CCX771/anti-CXCR7. Moreover, SNU449 and Huh7 cells exhibited matrix invasion in the presence of CXCL12 and CXCL11, a ligand exclusive to CXCR7. In conclusion, CXCR4 affects the prognosis of HCC patients but CXCR7 does not. Therefore, the CXCR4-CXCL12-CXCR7 axis plays a role in the interaction of HCC with the surrounding normal tissue and represents a suitable therapeutic target.

No MeSH data available.


Related in: MedlinePlus