Limits...
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.

Show MeSH

Related in: MedlinePlus

Expression of RANK and RANKL according to breast cancer subtype and correlation with Ki67. (a) Expression of RANKL by immunohistochemistry using the H-score (y axis) according to breast cancer subtypes in all patients, nonpregnant patients and pregnant patients. RANKL expression was higher in luminal A tumors (ER+, HER2–, Ki67 < 20%) compared with all other subtypes, particularly in pregnant patients (P <0.0001), than in nonpregnant patients (P = 0.32). (b) Negative correlation between Ki67 score by immunohistochemistry (y axis) and RANKL H-score (x axis) (P <0.0001, Pearson correlation = –0.29). (c) Expression of RANK by immunohistochemistry using the H-score (y axis) according to breast cancer subtypes in all patients, nonpregnant patients and pregnant patients. RANK expression was higher in triple-negative tumors compared with all other subtypes in all patients (P <0.0001), nonpregnant patients (P < 0.0001) and pregnant patients (P = 0.05). (d) Positive correlation between Ki67 score by immunohistochemistry (y axis) and RANK H-score (x axis) (P <0.0001, Pearson correlation = 0.37). ER, estrogen receptor; HER2, human epidermal growth factor receptor 2; 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
getmorefigures.php?uid=PMC4374174&req=5

Fig3: Expression of RANK and RANKL according to breast cancer subtype and correlation with Ki67. (a) Expression of RANKL by immunohistochemistry using the H-score (y axis) according to breast cancer subtypes in all patients, nonpregnant patients and pregnant patients. RANKL expression was higher in luminal A tumors (ER+, HER2–, Ki67 < 20%) compared with all other subtypes, particularly in pregnant patients (P <0.0001), than in nonpregnant patients (P = 0.32). (b) Negative correlation between Ki67 score by immunohistochemistry (y axis) and RANKL H-score (x axis) (P <0.0001, Pearson correlation = –0.29). (c) Expression of RANK by immunohistochemistry using the H-score (y axis) according to breast cancer subtypes in all patients, nonpregnant patients and pregnant patients. RANK expression was higher in triple-negative tumors compared with all other subtypes in all patients (P <0.0001), nonpregnant patients (P < 0.0001) and pregnant patients (P = 0.05). (d) Positive correlation between Ki67 score by immunohistochemistry (y axis) and RANK H-score (x axis) (P <0.0001, Pearson correlation = 0.37). ER, estrogen receptor; HER2, human epidermal growth factor receptor 2; RANK, receptor activator for nuclear factor κB; RANKL, receptor activator for nuclear factor κB ligand.

Mentions: Table 1 shows the association between RANK and RANKL expression on the primary tumor and clinicopathological features. For pregnant and nonpregnant patients, RANKL expression was higher in small (P = 0.01), well-differentiated (P <0.001) and PgR-positive tumors (P <0.001). On the contrary, higher RANK expression was observed in patients with poorly differentiated and hormone receptor-negative tumors (all P <0.001). RANKL expression was significantly higher in luminal A-like tumors (mean H-score: 45.45) compared with other subtypes, with the lowest expression observed in triple-negative subtypes (mean H-score: 0.23) (Figure 3a). Furthermore, the RANKL H-score was negatively correlated with Ki67 (P < 0.001) (Figure 3b). On the other hand, RANK expression was significantly higher in triple-negative tumors (mean H-score: 40.91) with the lowest expression observed in luminal A tumors (mean H-score: 4.68) (Figure 3c). A positive correlation was observed between RANK H-score and Ki67 (Figure 3d).Figure 3


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)

Expression of RANK and RANKL according to breast cancer subtype and correlation with Ki67. (a) Expression of RANKL by immunohistochemistry using the H-score (y axis) according to breast cancer subtypes in all patients, nonpregnant patients and pregnant patients. RANKL expression was higher in luminal A tumors (ER+, HER2–, Ki67 < 20%) compared with all other subtypes, particularly in pregnant patients (P <0.0001), than in nonpregnant patients (P = 0.32). (b) Negative correlation between Ki67 score by immunohistochemistry (y axis) and RANKL H-score (x axis) (P <0.0001, Pearson correlation = –0.29). (c) Expression of RANK by immunohistochemistry using the H-score (y axis) according to breast cancer subtypes in all patients, nonpregnant patients and pregnant patients. RANK expression was higher in triple-negative tumors compared with all other subtypes in all patients (P <0.0001), nonpregnant patients (P < 0.0001) and pregnant patients (P = 0.05). (d) Positive correlation between Ki67 score by immunohistochemistry (y axis) and RANK H-score (x axis) (P <0.0001, Pearson correlation = 0.37). ER, estrogen receptor; HER2, human epidermal growth factor receptor 2; 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

Fig3: Expression of RANK and RANKL according to breast cancer subtype and correlation with Ki67. (a) Expression of RANKL by immunohistochemistry using the H-score (y axis) according to breast cancer subtypes in all patients, nonpregnant patients and pregnant patients. RANKL expression was higher in luminal A tumors (ER+, HER2–, Ki67 < 20%) compared with all other subtypes, particularly in pregnant patients (P <0.0001), than in nonpregnant patients (P = 0.32). (b) Negative correlation between Ki67 score by immunohistochemistry (y axis) and RANKL H-score (x axis) (P <0.0001, Pearson correlation = –0.29). (c) Expression of RANK by immunohistochemistry using the H-score (y axis) according to breast cancer subtypes in all patients, nonpregnant patients and pregnant patients. RANK expression was higher in triple-negative tumors compared with all other subtypes in all patients (P <0.0001), nonpregnant patients (P < 0.0001) and pregnant patients (P = 0.05). (d) Positive correlation between Ki67 score by immunohistochemistry (y axis) and RANK H-score (x axis) (P <0.0001, Pearson correlation = 0.37). ER, estrogen receptor; HER2, human epidermal growth factor receptor 2; RANK, receptor activator for nuclear factor κB; RANKL, receptor activator for nuclear factor κB ligand.
Mentions: Table 1 shows the association between RANK and RANKL expression on the primary tumor and clinicopathological features. For pregnant and nonpregnant patients, RANKL expression was higher in small (P = 0.01), well-differentiated (P <0.001) and PgR-positive tumors (P <0.001). On the contrary, higher RANK expression was observed in patients with poorly differentiated and hormone receptor-negative tumors (all P <0.001). RANKL expression was significantly higher in luminal A-like tumors (mean H-score: 45.45) compared with other subtypes, with the lowest expression observed in triple-negative subtypes (mean H-score: 0.23) (Figure 3a). Furthermore, the RANKL H-score was negatively correlated with Ki67 (P < 0.001) (Figure 3b). On the other hand, RANK expression was significantly higher in triple-negative tumors (mean H-score: 40.91) with the lowest expression observed in luminal A tumors (mean H-score: 4.68) (Figure 3c). A positive correlation was observed between RANK H-score and Ki67 (Figure 3d).Figure 3

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