Quantitative Immunohistochemical Analysis Reveals Association between Sodium Iodide Symporter and Estrogen Receptor Expression in Breast Cancer

Abstract

BackgroundHuman sodium iodide symporter (hNIS) gene over-expression is under active consideration worldwide as an alternative target molecule for breast cancer (BC) diagnosis and targeted radio-iodine treatment. However, the field demands better stratified analysis of endogenous hNIS expression across major BC subtypes. Therefore, we have analyzed subtype-specific variation of hNIS overexpression in breast tumor tissue samples by immunohistochemistry (IHC) and also report the development of a homogeneous, quantitative analysis method of digital IHC images.MethodshNIS expression was analyzed from 108 BC tissue samples by IHC. Sub-cellular localization of hNIS protein was analyzed by dual immunofluorescence (IF) staining method using hNIS and HER2 antibodies. An ImageJ based two-step digital analysis method was developed and applied for the bias-free analysis of the images.ResultsStaining of the tumor samples show 70% cases are hNIS positive indicating high incidence of hNIS positive cases in BC. More importantly, a subtype specific analysis done for the first time shows that hNIS expression is overly dominated in estrogen receptor (ER) positive cases than the receptor negative cases. Further, 56% of the ER+ve, PgR+ve, HER2-ve and 36% of ER+ve, PgR+ve, HER2+ve cases show highest intensity staining equivalent to the thyroid tissue. A significant positive correlation is also observed between hNIS and estrogen receptor expression (p = 0.0033, CI = 95%) suggesting hNIS mediated targeted radio-iodine therapy procedures may benefit both ER+ve, PgR+ve, HER2–ve as well as HER2+ve cases. Further, in a few cases, hNIS and HER2 protein localization is demonstrated by overlapping membrane co-expression. ImageJ based image analysis method shows over 70% match with manual pathological scoring method.ConclusionThe study indicates a positive link between hNIS and ER expression in BC. The quantitative IHC image analysis method reported here will further help in patient stratification and potentially benefit global clinical assessment where hNIS mediated targeted 131I radio-ablative therapy is aimed.

Iodide transport and breast cancer

“…Expression of NIS does not appear to fluctuate between breast cancer tumour types, with tumours such as ductal carcinoma in situ and invasive carcinoma having similar NIS levels (Tazebay et al 2000, Wapnir et al 2003. The expression of NIS also appeared to be independent of breast cancer receptor status and has not been correlated to expression of TSHR, ER or PR in breast cancer (Oh et al 2005), although a more recent study did identify a correlation between ER and NIS expression (Chatterjee et al 2013).…”

Mig-6 deficiency cooperates with oncogenic Kras to promote mouse lung tumorigenesis

“…G. Representative results of phosphorylated p-mTOR (S2448) / mTOR (right panel) and the quantitative results (left panel) in mouse lungs. All quantifications are calculated from three mice per group (six regions for each mouse) using Image J [60]. Optical Density (OD) = log (max intensity/Mean intensity).…”

“…(A) Left panel: IHC staining results are the representative results for each group; Right panel: quantification results are calculated from three mice per group (six regions for each mouse) using Image J [60]. Optical Density (OD) = log (max intensity/Mean intensity).…”

The poly(A)-binding protein genes, EPAB, PABPC1, and PABPC3 are differentially expressed in infertile men with non-obstructive azoospermia

“…As a result, immunoexpression and localization of the PABPC1 and PABPC3 proteins in the NOA groups were examined with a bright-field microscopy (Zeiss). Also, we have further analyzed expression levels of the PABPC1 and PABPC3 proteins in all groups by using ImageJ software (National Institutes of Health, Bethesda, Maryland, USA), which confers us semiquantitative analysis of the immunohistochemical staining [33].…”