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β-Catenin immunocytochemical reactivity in cervicovaginal smears during regular menstrual cycles

Hanife Guler Donmez
Hanife Guler Donmez
   | 31 oct. 2020
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Article Category: Original article
Publié en ligne: 31 oct. 2020
Pages: 187 - 194
DOI: https://doi.org/10.1515/abm-2020-0027
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© 2020 Hanife Guler Donmez, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1

Immunocytochemical staining for β-catenin. A. Positive control, HeLa cells. Immunoreactivity for β-catenin is indicated by the dark-brown staining from oxidized diaminobenzidine chromogen used in the streptavidin–biotin method. The counterstain is Harris’ hematoxylin (magnification ×200, scale bar 200 μm, Olympus IX70 inverted microscope equipped with an Olympus DP71 digital camera). B. Negative control, cervicovaginal smear, without primary antibody. Only the Harris’ hematoxylin counterstain is seen (magnification ×400, scale bar 50 μm, Leica DM 4000B microscope equipped with a Leica DFC 370 digital camera).
Immunocytochemical staining for β-catenin. A. Positive control, HeLa cells. Immunoreactivity for β-catenin is indicated by the dark-brown staining from oxidized diaminobenzidine chromogen used in the streptavidin–biotin method. The counterstain is Harris’ hematoxylin (magnification ×200, scale bar 200 μm, Olympus IX70 inverted microscope equipped with an Olympus DP71 digital camera). B. Negative control, cervicovaginal smear, without primary antibody. Only the Harris’ hematoxylin counterstain is seen (magnification ×400, scale bar 50 μm, Leica DM 4000B microscope equipped with a Leica DFC 370 digital camera).

Figure 2

Wnt/β-catenin signaling pathway activity is suggested by immunoreactivity for β-catenin is indicated by the dark-brown staining from oxidized diaminobenzidine chromogen used in the streptavidin–biotin method. Harris’ hematoxylin counterstaining. A. Membranous and cytoplasmic immunoreactivity for β-catenin is depicted in the proliferative phase with the superficial cells being more prominent than intermediate cells. Activated Wnt/β-catenin signaling pathway is also seen in one superficial cell (arrow), which displays strong (+++) cytoplasmic immunoreactivity. B. Membranous and cytoplasmic immunoreactivity for β-catenin as seen in the secretory phase with the intermediate cells being more prominent than superficial cells. An activated Wnt/β-catenin signaling pathway is also observed in almost all intermediate cells (magnification ×400, scale bars 50 μm, Leica DM 4000B microscope equipped with a Leica DFC 370 digital camera).
Wnt/β-catenin signaling pathway activity is suggested by immunoreactivity for β-catenin is indicated by the dark-brown staining from oxidized diaminobenzidine chromogen used in the streptavidin–biotin method. Harris’ hematoxylin counterstaining. A. Membranous and cytoplasmic immunoreactivity for β-catenin is depicted in the proliferative phase with the superficial cells being more prominent than intermediate cells. Activated Wnt/β-catenin signaling pathway is also seen in one superficial cell (arrow), which displays strong (+++) cytoplasmic immunoreactivity. B. Membranous and cytoplasmic immunoreactivity for β-catenin as seen in the secretory phase with the intermediate cells being more prominent than superficial cells. An activated Wnt/β-catenin signaling pathway is also observed in almost all intermediate cells (magnification ×400, scale bars 50 μm, Leica DM 4000B microscope equipped with a Leica DFC 370 digital camera).

Figure 3

Comparison of immunocytochemical staining patterns between superficial, intermediate, and parabasal cells. Immunoreactivity for β-catenin is indicated by the dark-brown staining from oxidized diaminobenzidine chromogen used in the streptavidin–biotin method. Harris’ hematoxylin counterstaining. A. One of the two neighboring superficial cells show cytoplasmic immunoreactivity (arrow) and that Wnt/β-catenin signaling pathway is activated; however, the other cell is entirely unreactive. B. Membranous immunoreactivity is stronger in the intermediate cells than in the superficial cells. C. Strong cytoplasmic positivity and activated signaling are seen in an intermediate cell. D. Parabasal cells show stronger membranous and cytoplasmic immunoreactivity than superficial and intermediate cells (magnification ×1000), (scale bars 20 μm, Leica DM 4000B microscope equipped with a Leica DFC 370 digital camera).
Comparison of immunocytochemical staining patterns between superficial, intermediate, and parabasal cells. Immunoreactivity for β-catenin is indicated by the dark-brown staining from oxidized diaminobenzidine chromogen used in the streptavidin–biotin method. Harris’ hematoxylin counterstaining. A. One of the two neighboring superficial cells show cytoplasmic immunoreactivity (arrow) and that Wnt/β-catenin signaling pathway is activated; however, the other cell is entirely unreactive. B. Membranous immunoreactivity is stronger in the intermediate cells than in the superficial cells. C. Strong cytoplasmic positivity and activated signaling are seen in an intermediate cell. D. Parabasal cells show stronger membranous and cytoplasmic immunoreactivity than superficial and intermediate cells (magnification ×1000), (scale bars 20 μm, Leica DM 4000B microscope equipped with a Leica DFC 370 digital camera).

Figure 4

β-Catenin expression was reduced from parabasal to superficial layers in both the proliferative and secretory phases. H-scores were not normally distributed in all groups; thus, standard deviation (SD) was high (error bars indicate SD).
β-Catenin expression was reduced from parabasal to superficial layers in both the proliferative and secretory phases. H-scores were not normally distributed in all groups; thus, standard deviation (SD) was high (error bars indicate SD).

Comparing parameters in the menstrual cycle proliferative and secretory periods

Wnt/β-catenin signaling activity§Proliferative period n (%)Secretory period n (%)P
Active (n = 28)19 (73)9 (25)<0.001*†
Inactive (n = 34)7 (27)27 (75)
Total (n = 62)26 (100)36 (100)
Cellular localization of immunoreactivityMean ± SD||Mean ± SD||
Membranous β-catenin32.29 ± 34.418.89 ± 29.70.039*‡
Cytoplasmic β-catenin9.33 ± 12.752.62 ± 7.31<0.001**‡
Nuclear β-catenin1.07 ± 3.680.13 ± 0.800.033*‡
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