Table 1.
Primer pairs used in RT-PCR.
Table 2.
Summary of immunohistochemistry for AQP3 and AQP4 in thyroid tissues.
Figure 1.
Expression of AQP3 and AQP4 protein in normal and hyperplastic thyroid tissues/cells.
(A) Immunofluorescence double staining shows AQP3 (FITC: green) expressing in the cytoplasmic membranes of normal human C cells that are positive for calcitonin (TRCR: red), but not in follicular cells. (B) Immunoperoxidase staining also shows that AQP3 is positive only in C cells and not in follicular cells. (C) Immunofluorescence analysis exhibits AQP4 protein (FITC: green) in the cytoplasmic membranes of follicular cells. Nuclei are stained by DAPI (blue). (D) AQP4 protein positivity tends to be stronger in tall follicular cells of small follicles. Hyperplastic follicular cells in Graves’ disease (E) and multinodular goiter (F) display prominent positivity of AQP4 in their cytoplasmic membranes. (Magnification: A, C, 1000x; B, D, E, F, 400x).
Figure 2.
Immunohistochemical staining for AQP3 in neoplastic thyroid tissues.
Immunoreactivity of AQP3 is negative in follicular adenoma (A), follicular carcinoma (B), and papillary carcinoma (C). In contrast, AQP3 is clearly immunopositive in the cytoplasmic membranes of medullary carcinoma cells (D). (Magnification: 400x).
Figure 3.
Immunohistochemical staining for AQP4 in human neoplastic thyroid tissues.
AQP4 is diffusely immunopositive in neoplastic cells of follicular adenomas (A), follicular carcinomas (B), and papillary carcinomas (C), but negative in medullary carcinoma (D). (Magnification: 400x).
Figure 4.
Expression of AQP3 and AQP4 mRNA in human normal, hyperplastic and neoplastic thyroid tissues.
RT-PCR shows AQP3 mRNA only in medullary carcinomas (MC), and absent in normal thyroid tissues (NT), multinodular goiters (MG), follicular adenomas (FA), papillary carcinomas (PC), and undifferentiated carcinomas (UC). In contrast, AQP4 mRNA is demonstrated in NT, MG, FA, and PC, and negative in all UC and MC.
Figure 5.
Expression profiles of AQP3 and AQP4 in thyroid carcinoma cell lines.
(A) RT-PCR analysis shows that AQP3 mRNA is specifically identified in the TT cell line, whereas AQP4 is detected in the follicular cell-derived lines (KTC-1, TPC-1, WRO, UA-1, UA-2, and 8505C). (B) Western blotting confirms that AQP3 protein is seen in the TT cell line and not in the other cell lines. AQP4 protein is identified in five cell lines (KTC-1, TPC-1, UA-1, UA-2, and 8505C), while absent in three cell lines (TT, 8305C and WRO).
Figure 6.
Real time RT-PCR analysis for AQP3 mRNA in the TT cell line stimulated by fetal bovine serum (FBS) and calcium.
(A) AQP3 mRNA level is significantly increased by 5% and 10% FBS treatments. (B) AQP3 mRNA level is significantly up-regulated by calcium administration in a dose-dependent manner. Error bars, standard deviation. Significant difference, *P<0.05, **P<0.01.