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The prognostic significance of Wnt-5a expression in primary breast cancer is extended to premenopausal women.

Sand-Dejmek J, Ehrnström R, Berglund P, Andersson T, Ryden L - PLoS ONE (2013)

Bottom Line: Wnt-5a protein expression was reduced or lost in 146/361 of tumors and correlated to younger age, estrogen receptor (ER) negativity and triple-negative phenotype.In a subgroup analysis, Wnt-5a was significantly associated with better prognosis in patients with Luminal A tumors (p = 0.04).Conclusively, our results suggest that loss of Wnt-5a is a valuable prognostic marker in premenopausal breast cancer patients in particular in patients with ER-positive tumors and out-performed conventional prognostic factors in this subset of patients.

View Article: PubMed Central - PubMed

Affiliation: Experimental Pathology, Department of Laboratory Medicine, Lund University, Skåne University Hospital, Malmö, Sweden ; Surgery, Department of Clinical Sciences Malmö, Lund University, Skåne University Hospital, Malmö, Sweden.

ABSTRACT
Wnt-5a protein expression in primary tumors from unselected breast cancer patients has revealed a tumor suppressive function of the protein. However, in vitro experiments on human breast cancer cells have reported contradictory results, indicating both a tumor suppressive and promoting functions of Wnt-5a. This could be due to various functions of Wnt-5a in different subgroups of patients. The unselected cohorts analyzed to date for Wnt-5a protein expression contained few premenopausal patients. The aim of the present investigation was to evaluate the prognostic significance of Wnt-5a protein expression in a cohort of premenopausal women with comprehensive data on biomarkers, molecular subtypes and long-term outcome. In a randomized trial of adjuvant tamoxifen versus no adjuvant treatment, 564 premenopausal primary breast cancer patients were included. The median follow-up time was 14 years. A tumor tissue array was constructed and 361 samples were evaluated for Wnt-5a reactivity by immunohistochemistry. The primary end-point was recurrence-free survival. Wnt-5a protein expression was reduced or lost in 146/361 of tumors and correlated to younger age, estrogen receptor (ER) negativity and triple-negative phenotype. Wnt-5a was a prognostic factor in the whole cohort (p = 0.003). In patients with ER-positive tumors, Wnt-5a was an independent positive prognostic marker (HR 0.51 95% CI: 0.33-0.78 p = 0.002) and HER2 a negative prognostic marker (HR 2.84 95% CI: 1.51-5.31, p = 0.001) in a Cox multivariate analysis adjusted for standard prognostic markers and tamoxifen treatment. In the ER-negative subset, Wnt-5a added no prognostic information. In a subgroup analysis, Wnt-5a was significantly associated with better prognosis in patients with Luminal A tumors (p = 0.04). Conclusively, our results suggest that loss of Wnt-5a is a valuable prognostic marker in premenopausal breast cancer patients in particular in patients with ER-positive tumors and out-performed conventional prognostic factors in this subset of patients.

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Effect of exogenous ERα expression on the ability of Wnt-5a to decrease breast cancer cell invasion.A. ERα protein expression in MDA-MB-231 cells transfected with pcDNA3 empty vector (231 EV) or ERα expression plasmid (231 ERα). Lysate from the ER-positive breast cancer cell line T47D was included as a positive control. B. Maternal, EV transfected and the ER transfected MDA-MB-231 cells were tested for their expression of Wnt-5a in lanes 1, 3 and 4, respectively. As a positive control recombinant Wnt-5a was added to maternal MDA-MD-231 cell lysate and loaded in lane 2 as a positive control. C. Treatment with recombinant Wnt-5a or the Wnt-5a mimicking peptide Foxy5 significantly reduced the invasive capacity of both ER-negative and ER-positive MDA-MB-231 cells. Error bars, SEM. Paired t-test; **, P<0.01, ***, P<0.001.
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pone-0070890-g004: Effect of exogenous ERα expression on the ability of Wnt-5a to decrease breast cancer cell invasion.A. ERα protein expression in MDA-MB-231 cells transfected with pcDNA3 empty vector (231 EV) or ERα expression plasmid (231 ERα). Lysate from the ER-positive breast cancer cell line T47D was included as a positive control. B. Maternal, EV transfected and the ER transfected MDA-MB-231 cells were tested for their expression of Wnt-5a in lanes 1, 3 and 4, respectively. As a positive control recombinant Wnt-5a was added to maternal MDA-MD-231 cell lysate and loaded in lane 2 as a positive control. C. Treatment with recombinant Wnt-5a or the Wnt-5a mimicking peptide Foxy5 significantly reduced the invasive capacity of both ER-negative and ER-positive MDA-MB-231 cells. Error bars, SEM. Paired t-test; **, P<0.01, ***, P<0.001.

Mentions: In previous studies [7], [8] and in the present study we have demonstrated a correlation between the expression of Wnt-5a and ER and/or PR. This resulted in a design of both in vitro and in vivo experiments where reconstitution of Wnt-5a signaling was only tested in breast cancer cells lacking endogenous expression of both Wnt-5a and ER [16], [17]. The results from these studies revealed that Wnt5a primarily impaired the invasive properties of these cells whereas it only had a minor if any effect on cell proliferation [17]. Based on the present results we deemed it necessary to assess whether Wnt-5a affects breast cancer cell invasion differently in ER-positive compared to ER-negative breast cancer cells. We therefore performed Matrigel invasion assays with control transfected and ER-transfected MDA-MB-231 breast cancer cells. This approach enabled us to compare the possible difference in Wnt-5a-induced effect on cell invasion in ER expressing cells with those lacking ER expression in breast cancer cells with an identical phenotype. Transient ER transfection of ER-negative MDA-MB-231 cells resulted in an ER expression comparable to that of the ER-positive breast cancer cell line T47D (Figure 4A). Control experiments revealed that the Wnt-5a protein was neither expressed in maternal, EV transfected or ER transfected MDA-MB-231 cells (Figure 4B). As shown in figure 4C, stimulation with recombinant Wnt-5a or the Wnt-5a mimicking hexapeptide Foxy5 [16] did significantly decrease the invasive capacity of both ER-negative and ER-expressing cells, demonstrating that the ability of Wnt-5a signaling to impede breast cancer cell invasion is not affected by ER-status.


The prognostic significance of Wnt-5a expression in primary breast cancer is extended to premenopausal women.

Sand-Dejmek J, Ehrnström R, Berglund P, Andersson T, Ryden L - PLoS ONE (2013)

Effect of exogenous ERα expression on the ability of Wnt-5a to decrease breast cancer cell invasion.A. ERα protein expression in MDA-MB-231 cells transfected with pcDNA3 empty vector (231 EV) or ERα expression plasmid (231 ERα). Lysate from the ER-positive breast cancer cell line T47D was included as a positive control. B. Maternal, EV transfected and the ER transfected MDA-MB-231 cells were tested for their expression of Wnt-5a in lanes 1, 3 and 4, respectively. As a positive control recombinant Wnt-5a was added to maternal MDA-MD-231 cell lysate and loaded in lane 2 as a positive control. C. Treatment with recombinant Wnt-5a or the Wnt-5a mimicking peptide Foxy5 significantly reduced the invasive capacity of both ER-negative and ER-positive MDA-MB-231 cells. Error bars, SEM. Paired t-test; **, P<0.01, ***, P<0.001.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3750047&req=5

pone-0070890-g004: Effect of exogenous ERα expression on the ability of Wnt-5a to decrease breast cancer cell invasion.A. ERα protein expression in MDA-MB-231 cells transfected with pcDNA3 empty vector (231 EV) or ERα expression plasmid (231 ERα). Lysate from the ER-positive breast cancer cell line T47D was included as a positive control. B. Maternal, EV transfected and the ER transfected MDA-MB-231 cells were tested for their expression of Wnt-5a in lanes 1, 3 and 4, respectively. As a positive control recombinant Wnt-5a was added to maternal MDA-MD-231 cell lysate and loaded in lane 2 as a positive control. C. Treatment with recombinant Wnt-5a or the Wnt-5a mimicking peptide Foxy5 significantly reduced the invasive capacity of both ER-negative and ER-positive MDA-MB-231 cells. Error bars, SEM. Paired t-test; **, P<0.01, ***, P<0.001.
Mentions: In previous studies [7], [8] and in the present study we have demonstrated a correlation between the expression of Wnt-5a and ER and/or PR. This resulted in a design of both in vitro and in vivo experiments where reconstitution of Wnt-5a signaling was only tested in breast cancer cells lacking endogenous expression of both Wnt-5a and ER [16], [17]. The results from these studies revealed that Wnt5a primarily impaired the invasive properties of these cells whereas it only had a minor if any effect on cell proliferation [17]. Based on the present results we deemed it necessary to assess whether Wnt-5a affects breast cancer cell invasion differently in ER-positive compared to ER-negative breast cancer cells. We therefore performed Matrigel invasion assays with control transfected and ER-transfected MDA-MB-231 breast cancer cells. This approach enabled us to compare the possible difference in Wnt-5a-induced effect on cell invasion in ER expressing cells with those lacking ER expression in breast cancer cells with an identical phenotype. Transient ER transfection of ER-negative MDA-MB-231 cells resulted in an ER expression comparable to that of the ER-positive breast cancer cell line T47D (Figure 4A). Control experiments revealed that the Wnt-5a protein was neither expressed in maternal, EV transfected or ER transfected MDA-MB-231 cells (Figure 4B). As shown in figure 4C, stimulation with recombinant Wnt-5a or the Wnt-5a mimicking hexapeptide Foxy5 [16] did significantly decrease the invasive capacity of both ER-negative and ER-expressing cells, demonstrating that the ability of Wnt-5a signaling to impede breast cancer cell invasion is not affected by ER-status.

Bottom Line: Wnt-5a protein expression was reduced or lost in 146/361 of tumors and correlated to younger age, estrogen receptor (ER) negativity and triple-negative phenotype.In a subgroup analysis, Wnt-5a was significantly associated with better prognosis in patients with Luminal A tumors (p = 0.04).Conclusively, our results suggest that loss of Wnt-5a is a valuable prognostic marker in premenopausal breast cancer patients in particular in patients with ER-positive tumors and out-performed conventional prognostic factors in this subset of patients.

View Article: PubMed Central - PubMed

Affiliation: Experimental Pathology, Department of Laboratory Medicine, Lund University, Skåne University Hospital, Malmö, Sweden ; Surgery, Department of Clinical Sciences Malmö, Lund University, Skåne University Hospital, Malmö, Sweden.

ABSTRACT
Wnt-5a protein expression in primary tumors from unselected breast cancer patients has revealed a tumor suppressive function of the protein. However, in vitro experiments on human breast cancer cells have reported contradictory results, indicating both a tumor suppressive and promoting functions of Wnt-5a. This could be due to various functions of Wnt-5a in different subgroups of patients. The unselected cohorts analyzed to date for Wnt-5a protein expression contained few premenopausal patients. The aim of the present investigation was to evaluate the prognostic significance of Wnt-5a protein expression in a cohort of premenopausal women with comprehensive data on biomarkers, molecular subtypes and long-term outcome. In a randomized trial of adjuvant tamoxifen versus no adjuvant treatment, 564 premenopausal primary breast cancer patients were included. The median follow-up time was 14 years. A tumor tissue array was constructed and 361 samples were evaluated for Wnt-5a reactivity by immunohistochemistry. The primary end-point was recurrence-free survival. Wnt-5a protein expression was reduced or lost in 146/361 of tumors and correlated to younger age, estrogen receptor (ER) negativity and triple-negative phenotype. Wnt-5a was a prognostic factor in the whole cohort (p = 0.003). In patients with ER-positive tumors, Wnt-5a was an independent positive prognostic marker (HR 0.51 95% CI: 0.33-0.78 p = 0.002) and HER2 a negative prognostic marker (HR 2.84 95% CI: 1.51-5.31, p = 0.001) in a Cox multivariate analysis adjusted for standard prognostic markers and tamoxifen treatment. In the ER-negative subset, Wnt-5a added no prognostic information. In a subgroup analysis, Wnt-5a was significantly associated with better prognosis in patients with Luminal A tumors (p = 0.04). Conclusively, our results suggest that loss of Wnt-5a is a valuable prognostic marker in premenopausal breast cancer patients in particular in patients with ER-positive tumors and out-performed conventional prognostic factors in this subset of patients.

Show MeSH
Related in: MedlinePlus