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RANK-ligand (RANKL) expression in young breast cancer patients and during pregnancy.

Azim HA, Peccatori FA, Brohée S, Branstetter D, Loi S, Viale G, Piccart M, Dougall WC, Pruneri G, Sotiriou C - Breast Cancer Res. (2015)

Bottom Line: High RANKL expression within primary tumor was associated with pathways related to mammary gland development, bone resorption, T-cell proliferation and regulation of chemotaxis, while RANK expression was associated with immune response and proliferation pathways.At a median follow-up of 65 months, neither RANK nor RANKL expression within tumor was associated with disease free survival in pregnant or non-pregnant group.These results could guide further development of RANKL-targeted therapy.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: RANKL is important in mammary gland development during pregnancy and mediates the initiation and progression of progesterone-induced breast cancer. No clinical data are available on the effect of pregnancy on RANK/RANKL expression in young breast cancer patients.

Methods: We used our previously published dataset of 65 pregnant and 130 matched young breast cancer patients with full clinical, pathological, and survival information. 85% of patients had available transcriptomic data as well. RANK/RANKL expression by immunohistochemistry using H-score on the primary tumor and adjacent normal tissue was performed. We examined the difference in expression of RANK/RANKL between pregnant and non-pregnant patients and their association with clinicopathological features and prognosis. We also evaluated genes and pathways associated with RANK/RANKL expression on primary tumors.

Results: RANKL but not RANK expression was more prevalent in the pregnant group, both on the tumor and adjacent normal tissue, independent of other clinicopathological factors (both P <0.001). 18.7% of pregnant and 5.3% of non-pregnant patients had tumors showing ≥10% of cells with 3+ RANKL expression. RANKL expression was significantly higher in progesterone receptor-positive, and luminal A-like tumors, with negative correlation with Ki-67 (all P <0.001). On the contrary, RANK expression was higher in triple negative tumors (P <0.001). Using false discovery rate <0.05, 151 and 1,207 genes were significantly correlated with tumor-expressed RANKL and RANK expression by immunohistochemistry, respectively. High RANKL expression within primary tumor was associated with pathways related to mammary gland development, bone resorption, T-cell proliferation and regulation of chemotaxis, while RANK expression was associated with immune response and proliferation pathways. At a median follow-up of 65 months, neither RANK nor RANKL expression within tumor was associated with disease free survival in pregnant or non-pregnant group.

Conclusions: Pregnancy increases RANKL expression both in normal breast and primary tumors. These results could guide further development of RANKL-targeted therapy.

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Correlation between RANK and RANKL H-score and mRNA levels. (a) Positive correlation between RANKL mRNA expression (y axis) and RANKL H-score by immunohistochemistry (x axis) (P <0.0001, Pearson correlation = 0.89). (b) Positive correlation between RANK mRNA expression (y axis) and RANK H-score by immunohistochemistry (x axis) (P = 0.01, Pearson correlation = 0.19). RANK, receptor activator for nuclear factor κB; RANKL, receptor activator for nuclear factor κB ligand.
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Fig4: Correlation between RANK and RANKL H-score and mRNA levels. (a) Positive correlation between RANKL mRNA expression (y axis) and RANKL H-score by immunohistochemistry (x axis) (P <0.0001, Pearson correlation = 0.89). (b) Positive correlation between RANK mRNA expression (y axis) and RANK H-score by immunohistochemistry (x axis) (P = 0.01, Pearson correlation = 0.19). RANK, receptor activator for nuclear factor κB; RANKL, receptor activator for nuclear factor κB ligand.

Mentions: Using false discovery rate <0.05, out of 18,665 evaluated genes 1,207 genes and 151 genes were significantly correlated (either positively or negatively) with RANK and RANKL H-score, respectively (Additional file 4). Almost perfect correlation was observed between tumor RANKL H-score and mRNA levels (r = 0.89, P < 0.001) (Figure 4a). A weaker correlation, albeit significant, was observed between tumor RANK H-score and mRNA levels (r = 0.19, P = 0.012) (Figure 4b). GSEA showed that tumors expressing high levels of RANKL by immunohistochemistry had activated pathways; of particular relevance were those related to bone resorption, mammary gland development, regulation of chemotaxis and T-cell proliferation. On the other hand, high RANK expression was associated with several activated pathways, of relevance to immune response, and proliferation-related pathways (Additional file 5). These results were consistent even after adjusting the analysis for pregnancy status.Figure 4


RANK-ligand (RANKL) expression in young breast cancer patients and during pregnancy.

Azim HA, Peccatori FA, Brohée S, Branstetter D, Loi S, Viale G, Piccart M, Dougall WC, Pruneri G, Sotiriou C - Breast Cancer Res. (2015)

Correlation between RANK and RANKL H-score and mRNA levels. (a) Positive correlation between RANKL mRNA expression (y axis) and RANKL H-score by immunohistochemistry (x axis) (P <0.0001, Pearson correlation = 0.89). (b) Positive correlation between RANK mRNA expression (y axis) and RANK H-score by immunohistochemistry (x axis) (P = 0.01, Pearson correlation = 0.19). RANK, receptor activator for nuclear factor κB; RANKL, receptor activator for nuclear factor κB ligand.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4374174&req=5

Fig4: Correlation between RANK and RANKL H-score and mRNA levels. (a) Positive correlation between RANKL mRNA expression (y axis) and RANKL H-score by immunohistochemistry (x axis) (P <0.0001, Pearson correlation = 0.89). (b) Positive correlation between RANK mRNA expression (y axis) and RANK H-score by immunohistochemistry (x axis) (P = 0.01, Pearson correlation = 0.19). RANK, receptor activator for nuclear factor κB; RANKL, receptor activator for nuclear factor κB ligand.
Mentions: Using false discovery rate <0.05, out of 18,665 evaluated genes 1,207 genes and 151 genes were significantly correlated (either positively or negatively) with RANK and RANKL H-score, respectively (Additional file 4). Almost perfect correlation was observed between tumor RANKL H-score and mRNA levels (r = 0.89, P < 0.001) (Figure 4a). A weaker correlation, albeit significant, was observed between tumor RANK H-score and mRNA levels (r = 0.19, P = 0.012) (Figure 4b). GSEA showed that tumors expressing high levels of RANKL by immunohistochemistry had activated pathways; of particular relevance were those related to bone resorption, mammary gland development, regulation of chemotaxis and T-cell proliferation. On the other hand, high RANK expression was associated with several activated pathways, of relevance to immune response, and proliferation-related pathways (Additional file 5). These results were consistent even after adjusting the analysis for pregnancy status.Figure 4

Bottom Line: High RANKL expression within primary tumor was associated with pathways related to mammary gland development, bone resorption, T-cell proliferation and regulation of chemotaxis, while RANK expression was associated with immune response and proliferation pathways.At a median follow-up of 65 months, neither RANK nor RANKL expression within tumor was associated with disease free survival in pregnant or non-pregnant group.These results could guide further development of RANKL-targeted therapy.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: RANKL is important in mammary gland development during pregnancy and mediates the initiation and progression of progesterone-induced breast cancer. No clinical data are available on the effect of pregnancy on RANK/RANKL expression in young breast cancer patients.

Methods: We used our previously published dataset of 65 pregnant and 130 matched young breast cancer patients with full clinical, pathological, and survival information. 85% of patients had available transcriptomic data as well. RANK/RANKL expression by immunohistochemistry using H-score on the primary tumor and adjacent normal tissue was performed. We examined the difference in expression of RANK/RANKL between pregnant and non-pregnant patients and their association with clinicopathological features and prognosis. We also evaluated genes and pathways associated with RANK/RANKL expression on primary tumors.

Results: RANKL but not RANK expression was more prevalent in the pregnant group, both on the tumor and adjacent normal tissue, independent of other clinicopathological factors (both P <0.001). 18.7% of pregnant and 5.3% of non-pregnant patients had tumors showing ≥10% of cells with 3+ RANKL expression. RANKL expression was significantly higher in progesterone receptor-positive, and luminal A-like tumors, with negative correlation with Ki-67 (all P <0.001). On the contrary, RANK expression was higher in triple negative tumors (P <0.001). Using false discovery rate <0.05, 151 and 1,207 genes were significantly correlated with tumor-expressed RANKL and RANK expression by immunohistochemistry, respectively. High RANKL expression within primary tumor was associated with pathways related to mammary gland development, bone resorption, T-cell proliferation and regulation of chemotaxis, while RANK expression was associated with immune response and proliferation pathways. At a median follow-up of 65 months, neither RANK nor RANKL expression within tumor was associated with disease free survival in pregnant or non-pregnant group.

Conclusions: Pregnancy increases RANKL expression both in normal breast and primary tumors. These results could guide further development of RANKL-targeted therapy.

Show MeSH
Related in: MedlinePlus