The sodium iodide symporter (NIS) and potential regulators in normal, benign and malignant human breast tissue

James Ryan, Catherine E Curran, Emer Hennessy, John Newell, John C Morris, Michael J Kerin, Roisin M Dwyer, James Ryan, Catherine E Curran, Emer Hennessy, John Newell, John C Morris, Michael J Kerin, Roisin M Dwyer

Abstract

Introduction: The presence, relevance and regulation of the Sodium Iodide Symporter (NIS) in human mammary tissue remains poorly understood. This study aimed to quantify relative expression of NIS and putative regulators in human breast tissue, with relationships observed further investigated in vitro.

Methods: Human breast tissue specimens (malignant n = 75, normal n = 15, fibroadenoma n = 10) were analysed by RQ-PCR targeting NIS, receptors for retinoic acid (RARα, RARβ), oestrogen (ERα), thyroid hormones (THRα, THRβ), and also phosphoinositide-3-kinase (PI3K). Breast cancer cells were treated with Retinoic acid (ATRA), Estradiol and Thyroxine individually and in combination followed by analysis of changes in NIS expression.

Results: The lowest levels of NIS were detected in normal tissue (Mean(SEM) 0.70(0.12) Log(10) Relative Quantity (RQ)) with significantly higher levels observed in fibroadenoma (1.69(0.21) Log(10)RQ, p<0.005) and malignant breast tissue (1.18(0.07) Log(10)RQ, p<0.05). Significant positive correlations were observed between human NIS and ERα (r = 0.22, p<0.05) and RARα (r = 0.29, p<0.005), with the strongest relationship observed between NIS and RARβ (r = 0.38, p<0.0001). An inverse relationship between NIS and PI3K expression was also observed (r = 0.21, p<0.05). In vitro, ATRA, Estradiol and Thyroxine individually stimulated significant increases in NIS expression (range 6-16 fold), while ATRA and Thyroxine combined caused the greatest increase (range 16-26 fold).

Conclusion: Although NIS expression is significantly higher in malignant compared to normal breast tissue, the highest level was detected in fibroadenoma. The data presented supports a role for retinoic acid and estradiol in mammary NIS regulation in vivo, and also highlights potential thyroidal regulation of mammary NIS mediated by thyroid hormones.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. NIS expression in normal, fibroadenoma…
Figure 1. NIS expression in normal, fibroadenoma (“Fibro”) and malignant breast tissue.
Results have been normalised to the endogenous controls PPIA and MRPL19 and expressed relative to the lowest detectable sample. Note: Results in breast cancer patients are also stratified by epithelial subtype.
Figure 2. Immunohistochemical detection of NIS protein…
Figure 2. Immunohistochemical detection of NIS protein expression in 5 µM sections of selected tissues from patient cohort.
(a) Thyroid tissue (positive control) (400×), (b) Antibody free Fibroadenoma (negative control) (200×), (c) Fibroadenoma (400×). Also shown are breast cancer epithelial subtypes Luminal A (d), Luminal B (e), Her2 (f) and Basal (g) at 400× magnification.
Figure 3. RARα and RARβ expression in…
Figure 3. RARα and RARβ expression in breast tissue samples, and their relationship with NIS.
(a) RARα and RARβ expression in normal, malignant and fibroadenoma breast tissue. Results have been normalised to the endogenous controls PPIA and MRPL19 and expressed relative to the lowest detectable sample. Correlations of NIS gene expression with RARα (b) and RARβ (c) with r and p values shown (* one outlier removed). (d) THRα and THRβ expression in normal, malignant and fibroadenoma breast tissue.
Figure 4. ERα and PI3K expression in…
Figure 4. ERα and PI3K expression in breast tissue samples and their relationship with NIS.
Expression of ERα (a) and PI3K (b) in normal, malignant and fibroadenoma breast tissue. Results have been normalised to the endogenous controls PPIA and MRPL19 and expressed relative to the lowest detectable sample. Correlation of NIS gene expression with ERα (c) and PI3K (d) with r and p values shown.
Figure 5. Effect of β-estradiol (E 2…
Figure 5. Effect of β-estradiol (E2), all trans Retinoic Acid (ATRA) and L-thyroxine (T4) on NIS expression in breast cancer cell lines.
Effect of indivdual stimulants on NIS expression in (a) T47D and (b) SkBr3 cell lines. Results have been normalised to the endogenous controls PPIA and MRPL19 and expressed relative to diluent controls, with the baseline representing 2−(ddCt diluent control). (c) NIS expression in T47D and SkBr3 cells following 24 hr stimulation with optimal concentrations of ligands alone and in combination. (d) Correlation of RARβ with THRβ gene expression within patient cohort with r and p values shown.

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