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Expression of pendrin in benign and malignant human thyroid tissues.

Skubis-Zegadło J, Nikodemska A, Przytuła E, Mikula M, Bardadin K, Ostrowski J, Wenzel BE, Czarnocka B - Br. J. Cancer (2005)

Bottom Line: Pendrin protein was detected in 73.3 and 76.7% of the follicular (FTC) and papillary (PTC) thyroid carcinomas, respectively, where pendrin was solely localised inside the cytoplasm.An extensive intracellular immunostaining of pendrin was observed in six out of 11 (54.5%) of positive FTCs and 19 out of 23 (82%) of PTCs.In conclusion, pendrin is expressed in the majority of differentiated thyroid tumours with high individual variability but its targeting to the apical cell membrane is affected.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Medical Centre for Postgraduate Education, Marymoncka 99, 01-813 Warsaw, Poland.

ABSTRACT
The Pendred syndrome gene (PDS) encodes a transmembrane protein, pendrin, which is expressed in follicular thyroid cells and participates in the apical iodide transport. Pendrin expression has been studied in various thyroid neoplasms by means of immunohistochemistry (IHC), Western blot and RT-quantitative real-time PCR. The expression was related to the functional activity of the thyroid tissue. Follicular cells of normal, nodular goitre and Graves' disease tissues express pendrin at the apical pole of the thyrocytes. In follicular adenomas, pendrin was detected in cell membranes and cytoplasm simultaneously in 10 out of 15 cases. Pendrin protein was detected in 73.3 and 76.7% of the follicular (FTC) and papillary (PTC) thyroid carcinomas, respectively, where pendrin was solely localised inside the cytoplasm. An extensive intracellular immunostaining of pendrin was observed in six out of 11 (54.5%) of positive FTCs and 19 out of 23 (82%) of PTCs. Focal reactivity was detected in one follicular- and three papillary carcinomas, whereas pendrin protein was absent in three of 15 FTC and four of 30 PTC; mRNA of pendrin was detected in 92.4% of thyroid tumours. The relative mRNA expression of pendrin was lower in cancers than in normal thyroid tissues (P<0.001). The pendrin protein level was found to parallel its mRNA expression, which was not, however, related to the tumour size and tumour stage. In conclusion, pendrin is expressed in the majority of differentiated thyroid tumours with high individual variability but its targeting to the apical cell membrane is affected.

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Immunohistochemical analysis of expression and cellular distribution of pendrin in normal and pathological thyroid tissues. (A) Normal thyroid tissue; the apical staining is heterogeneous inside and between follicles (original magnification × 400; inset – peptide competition test control); (B) immunostaining for pendrin in NG. The staining is restricted to the small proliferating follicles (original magnification × 200; inset – peptide competition test control); (C) Graves' tissue with distinct apical staining pattern, especially observed in small follicles (original magnification × 200; inset – peptide competition test control); (D) immunostaining of pendrin in Graves' tissue by confocal microscopy, FITC-labelled anti-rabbit secondary antibody were used. (E) Follicular adenoma; immunostaining shift to the cytoplasm is observed (original magnification × 400; inset – peptide competition test control); (F) Follicular carcinoma; strong intracellular staining (original magnification × 400); (G) Follicular carcinoma; moderate intracytoplasmic stain (original magnification × 400); (H) immunofluorescence of FTC with strong intracytoplasmic pendrin expression (original magnification × 40); (I) Papillary carcinoma; strong intracellular staining (original magnification × 400; inset – peptide precipitation test control); (J) Papillary thyroid carcinoma; moderate cytoplasmic staining (original magnification × 200); (K) immunofluorescence of PTC with strong pendrin expression (original magnification × 40); (L) immunofluorescence of PTC with moderate pendrin expression (original magnification × 40).
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fig3: Immunohistochemical analysis of expression and cellular distribution of pendrin in normal and pathological thyroid tissues. (A) Normal thyroid tissue; the apical staining is heterogeneous inside and between follicles (original magnification × 400; inset – peptide competition test control); (B) immunostaining for pendrin in NG. The staining is restricted to the small proliferating follicles (original magnification × 200; inset – peptide competition test control); (C) Graves' tissue with distinct apical staining pattern, especially observed in small follicles (original magnification × 200; inset – peptide competition test control); (D) immunostaining of pendrin in Graves' tissue by confocal microscopy, FITC-labelled anti-rabbit secondary antibody were used. (E) Follicular adenoma; immunostaining shift to the cytoplasm is observed (original magnification × 400; inset – peptide competition test control); (F) Follicular carcinoma; strong intracellular staining (original magnification × 400); (G) Follicular carcinoma; moderate intracytoplasmic stain (original magnification × 400); (H) immunofluorescence of FTC with strong intracytoplasmic pendrin expression (original magnification × 40); (I) Papillary carcinoma; strong intracellular staining (original magnification × 400; inset – peptide precipitation test control); (J) Papillary thyroid carcinoma; moderate cytoplasmic staining (original magnification × 200); (K) immunofluorescence of PTC with strong pendrin expression (original magnification × 40); (L) immunofluorescence of PTC with moderate pendrin expression (original magnification × 40).

Mentions: The result of immunohistochemical analyses of the pendrin in 155 thyroid tissues are summarised in Table 1 and are shown in Figure 3. In the normal thyroid, GD and NG tissues, the pendrin staining was limited to the apical cell membrane (Figure 3A). One of 15 follicular adenomas showed exclusively membranous pendrin expression. In 10 of 15 (66.7%) adenomas, the expression of pendrin protein was detected simultaneously in the cytoplasm and at the apical cell membrane with extensive and moderate immunostaining detected in half of all cases. Four of 15 adenomas (26.7%) expressed pendrin protein in the cytoplasm only with strong immunoreactivity detected in three cases. In a series of differentiated thyroid carcinomas, 73.3% (11 out of 15) of FTC and 76.6% (23 out of 30) of PTC clearly showed pendrin protein expression. Out of 11 follicular carcinomas, 10 (91%) expressed pendrin protein exclusively in the cytoplasm, and one both in the cytoplasm and at the apical cell membrane. In six of 10 FTC, the immunostaining for pendrin was intensive and homogenously distributed through the tumoral area, and in the remaining four cases intermediate reactivity was observed. Three (20%) follicular carcinomas were negative and one exhibited focal pendrin immunostaining. Of 30 papillary carcinomas, 23 showed pendrin protein expression. In all positive tumours, the pendrin protein was localised exclusively in the cytoplasm. Out of these 23 cases, 19 PTC showed an intensive homogenous pendrin expression, whereas in the remaining four cases moderate immunostaining was found. Four (13.3%) papillary carcinomas were negative for pendrin staining, and three showed focal reactivity (Table 1). Pendrin protein immunodetection on paraffin-embedded sections is shown in Figure 3. Affinity-purified anti-pendrin antibodies specifically labelled pendrin located at the apical plasma membranes of follicular cells of the normal thyroid (Figure 3A), NG (Figure 3B) and GD (Figure 3C) tissues. In the normal thyroid, the apical labelling was not uniform in both follicles and epithelial cells. These stained cells likely correspond to functionally active cells and follicles. In NGs (Figure 3B) and GD thyroids (Figure 3C), immunostaining varied from one follicle to another and was related to the size of the follicles (Figure 3B and C). The majority of positive cells were in small follicles and in follicles with proliferative activity, whereas in large follicles the proportion of positive cells was decreased (Figure 3B and C). In follicular adenomas, pendrin-positive immunostaining was observed at both, the apical plasma membranes and in the cytoplasm of 14 out of 15 cases (Figure 3E). In FTC and PTC, the pendrin protein immunostaining of variable intensity was uniformly distributed throughout the tumoral area and generally located in the cytoplasm and probably membrane-associated. Figure 3F+I shows cases of and PTCs with an extensive intracellular pendrin expression, and Figure 3G+J illustrates follicular and papillary carcinomas with moderate pendrin expression. The pattern of reactivity was confirmed for all tissues analysed by confocal fluorescent microscopy (Figure 3D – Graves' tissue; H – FTC; K+L – PTC carcinomas). There was no relationship between pendrin expression or pendrin localisation and tumour stages (TNM), or tumour size (Table 2). The analysed anaplastic carcinomas with characteristics of the fully dedifferentiated tumours and no expression of thyroid-specific genes as well as the few cases of medullary carcinomas derived from C cells of neuronal crest origin were negative for PDS gene transcript and pendrin protein in IHC and Western blots.


Expression of pendrin in benign and malignant human thyroid tissues.

Skubis-Zegadło J, Nikodemska A, Przytuła E, Mikula M, Bardadin K, Ostrowski J, Wenzel BE, Czarnocka B - Br. J. Cancer (2005)

Immunohistochemical analysis of expression and cellular distribution of pendrin in normal and pathological thyroid tissues. (A) Normal thyroid tissue; the apical staining is heterogeneous inside and between follicles (original magnification × 400; inset – peptide competition test control); (B) immunostaining for pendrin in NG. The staining is restricted to the small proliferating follicles (original magnification × 200; inset – peptide competition test control); (C) Graves' tissue with distinct apical staining pattern, especially observed in small follicles (original magnification × 200; inset – peptide competition test control); (D) immunostaining of pendrin in Graves' tissue by confocal microscopy, FITC-labelled anti-rabbit secondary antibody were used. (E) Follicular adenoma; immunostaining shift to the cytoplasm is observed (original magnification × 400; inset – peptide competition test control); (F) Follicular carcinoma; strong intracellular staining (original magnification × 400); (G) Follicular carcinoma; moderate intracytoplasmic stain (original magnification × 400); (H) immunofluorescence of FTC with strong intracytoplasmic pendrin expression (original magnification × 40); (I) Papillary carcinoma; strong intracellular staining (original magnification × 400; inset – peptide precipitation test control); (J) Papillary thyroid carcinoma; moderate cytoplasmic staining (original magnification × 200); (K) immunofluorescence of PTC with strong pendrin expression (original magnification × 40); (L) immunofluorescence of PTC with moderate pendrin expression (original magnification × 40).
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fig3: Immunohistochemical analysis of expression and cellular distribution of pendrin in normal and pathological thyroid tissues. (A) Normal thyroid tissue; the apical staining is heterogeneous inside and between follicles (original magnification × 400; inset – peptide competition test control); (B) immunostaining for pendrin in NG. The staining is restricted to the small proliferating follicles (original magnification × 200; inset – peptide competition test control); (C) Graves' tissue with distinct apical staining pattern, especially observed in small follicles (original magnification × 200; inset – peptide competition test control); (D) immunostaining of pendrin in Graves' tissue by confocal microscopy, FITC-labelled anti-rabbit secondary antibody were used. (E) Follicular adenoma; immunostaining shift to the cytoplasm is observed (original magnification × 400; inset – peptide competition test control); (F) Follicular carcinoma; strong intracellular staining (original magnification × 400); (G) Follicular carcinoma; moderate intracytoplasmic stain (original magnification × 400); (H) immunofluorescence of FTC with strong intracytoplasmic pendrin expression (original magnification × 40); (I) Papillary carcinoma; strong intracellular staining (original magnification × 400; inset – peptide precipitation test control); (J) Papillary thyroid carcinoma; moderate cytoplasmic staining (original magnification × 200); (K) immunofluorescence of PTC with strong pendrin expression (original magnification × 40); (L) immunofluorescence of PTC with moderate pendrin expression (original magnification × 40).
Mentions: The result of immunohistochemical analyses of the pendrin in 155 thyroid tissues are summarised in Table 1 and are shown in Figure 3. In the normal thyroid, GD and NG tissues, the pendrin staining was limited to the apical cell membrane (Figure 3A). One of 15 follicular adenomas showed exclusively membranous pendrin expression. In 10 of 15 (66.7%) adenomas, the expression of pendrin protein was detected simultaneously in the cytoplasm and at the apical cell membrane with extensive and moderate immunostaining detected in half of all cases. Four of 15 adenomas (26.7%) expressed pendrin protein in the cytoplasm only with strong immunoreactivity detected in three cases. In a series of differentiated thyroid carcinomas, 73.3% (11 out of 15) of FTC and 76.6% (23 out of 30) of PTC clearly showed pendrin protein expression. Out of 11 follicular carcinomas, 10 (91%) expressed pendrin protein exclusively in the cytoplasm, and one both in the cytoplasm and at the apical cell membrane. In six of 10 FTC, the immunostaining for pendrin was intensive and homogenously distributed through the tumoral area, and in the remaining four cases intermediate reactivity was observed. Three (20%) follicular carcinomas were negative and one exhibited focal pendrin immunostaining. Of 30 papillary carcinomas, 23 showed pendrin protein expression. In all positive tumours, the pendrin protein was localised exclusively in the cytoplasm. Out of these 23 cases, 19 PTC showed an intensive homogenous pendrin expression, whereas in the remaining four cases moderate immunostaining was found. Four (13.3%) papillary carcinomas were negative for pendrin staining, and three showed focal reactivity (Table 1). Pendrin protein immunodetection on paraffin-embedded sections is shown in Figure 3. Affinity-purified anti-pendrin antibodies specifically labelled pendrin located at the apical plasma membranes of follicular cells of the normal thyroid (Figure 3A), NG (Figure 3B) and GD (Figure 3C) tissues. In the normal thyroid, the apical labelling was not uniform in both follicles and epithelial cells. These stained cells likely correspond to functionally active cells and follicles. In NGs (Figure 3B) and GD thyroids (Figure 3C), immunostaining varied from one follicle to another and was related to the size of the follicles (Figure 3B and C). The majority of positive cells were in small follicles and in follicles with proliferative activity, whereas in large follicles the proportion of positive cells was decreased (Figure 3B and C). In follicular adenomas, pendrin-positive immunostaining was observed at both, the apical plasma membranes and in the cytoplasm of 14 out of 15 cases (Figure 3E). In FTC and PTC, the pendrin protein immunostaining of variable intensity was uniformly distributed throughout the tumoral area and generally located in the cytoplasm and probably membrane-associated. Figure 3F+I shows cases of and PTCs with an extensive intracellular pendrin expression, and Figure 3G+J illustrates follicular and papillary carcinomas with moderate pendrin expression. The pattern of reactivity was confirmed for all tissues analysed by confocal fluorescent microscopy (Figure 3D – Graves' tissue; H – FTC; K+L – PTC carcinomas). There was no relationship between pendrin expression or pendrin localisation and tumour stages (TNM), or tumour size (Table 2). The analysed anaplastic carcinomas with characteristics of the fully dedifferentiated tumours and no expression of thyroid-specific genes as well as the few cases of medullary carcinomas derived from C cells of neuronal crest origin were negative for PDS gene transcript and pendrin protein in IHC and Western blots.

Bottom Line: Pendrin protein was detected in 73.3 and 76.7% of the follicular (FTC) and papillary (PTC) thyroid carcinomas, respectively, where pendrin was solely localised inside the cytoplasm.An extensive intracellular immunostaining of pendrin was observed in six out of 11 (54.5%) of positive FTCs and 19 out of 23 (82%) of PTCs.In conclusion, pendrin is expressed in the majority of differentiated thyroid tumours with high individual variability but its targeting to the apical cell membrane is affected.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Medical Centre for Postgraduate Education, Marymoncka 99, 01-813 Warsaw, Poland.

ABSTRACT
The Pendred syndrome gene (PDS) encodes a transmembrane protein, pendrin, which is expressed in follicular thyroid cells and participates in the apical iodide transport. Pendrin expression has been studied in various thyroid neoplasms by means of immunohistochemistry (IHC), Western blot and RT-quantitative real-time PCR. The expression was related to the functional activity of the thyroid tissue. Follicular cells of normal, nodular goitre and Graves' disease tissues express pendrin at the apical pole of the thyrocytes. In follicular adenomas, pendrin was detected in cell membranes and cytoplasm simultaneously in 10 out of 15 cases. Pendrin protein was detected in 73.3 and 76.7% of the follicular (FTC) and papillary (PTC) thyroid carcinomas, respectively, where pendrin was solely localised inside the cytoplasm. An extensive intracellular immunostaining of pendrin was observed in six out of 11 (54.5%) of positive FTCs and 19 out of 23 (82%) of PTCs. Focal reactivity was detected in one follicular- and three papillary carcinomas, whereas pendrin protein was absent in three of 15 FTC and four of 30 PTC; mRNA of pendrin was detected in 92.4% of thyroid tumours. The relative mRNA expression of pendrin was lower in cancers than in normal thyroid tissues (P<0.001). The pendrin protein level was found to parallel its mRNA expression, which was not, however, related to the tumour size and tumour stage. In conclusion, pendrin is expressed in the majority of differentiated thyroid tumours with high individual variability but its targeting to the apical cell membrane is affected.

Show MeSH
Related in: MedlinePlus