Table 1.
Characteristic of patient cohort.
Figure 1.
Images of hNIS staining pattern in major subtypes of breast cancer.
A) Tumor adjacent normal breast tissue showing negative expression of hNIS. B) Salivary gland tissue section showing membrane staining for hNIS. C) Thyroid papillary carcinoma tissue section as positive control. D-F) hNIS expression in ER+ve, PgR+ve, HER2-ve subtype representing 0, 1+ and 2+ score respectively. G-I) hNIS expression in ER+ve, PgR+ve, HER2+ve subtype with 0, 1+ and 2+ score respectively. J-L) hNIS expression in ER-ve, PgR-ve, HER2+ve subtype with 0, 1+ and 2+ score respectively. M-O) hNIS expression in ER-ve, PgR-ve, HER2-ve subtype with 0, 1+ and 2+ scores respectively. All images are captured at 20X magnification.
Figure 2.
hNIS expression varies in different patient cohorts.
A) Chart representing positive and negative hNIS expression in 108 cases of breast cancer representing major subtypes. B) hNIS positive expression is compared between the ER-ve and ER+ve patient samples indicating significant association of hNIS expression in ER+ve group. C) hNIS positive expression is compared between HER2–ve and HER2+ve patient samples indicating no association. D) hNIS positive expression in pre- and post-menopausal group of patients indicating significant higher hNIS expression in pre-menopausal women.
Figure 3.
Immunofluorescence staining reveals hNIS and HER2 receptor overlaps at the tumor cell membrane in few cases.
Human patient tissue samples stained with hNIS and HER2 specific antibody followed by detection with Alexafluor488 (green) and Dylight633 (red) labeled secondary antibodies respectively. A, E) DAB stained tissue sections from two representative patient samples showing similar intracellular expression of hNIS. B, F) Dual IF stained confocal overlay images of the same patient samples as in A and E respectively. Upper panel clearly showed good overlapping membrane expression of HER2 and hNIS staining. C, G) Corresponding images of HER2 staining in red channel and D, H) images of hNIS staining in green channel.
Figure 4.
Diagrammatic representation of different zones classified for scoring DAB stained IHC images.
A) IHC image, their DAB color de-convoluted image, reference histogram profile and pixel analysis data table for each are represented for cases scored as 2+, 1+ and 0. On the histogram profile, subdivision of zones based on pixel intensities are marked with colored boxes as zone 2 (61–120) with pink, zone 3 (121–170) with yellow and zone 4 (171–230) with light yellow. B) software-based analysis data set of the papillary thyroid carcinoma sample representing positive (2+) score and tumor adjacent normal sample representing negative score are represented.
Figure 5.
Subtype specific hNIS scoring analysis in breast cancer. A
) Chart representing IHC scoring for hNIS in various breast cancer subtypes by pathological analysis. B) IHC scoring for hNIS of the same dataset by semi-automated process developed. On an average 68% match and 75% match with 2+ score is observed between the two scoring methods.