Open Access

Human papillomavirus (HPV) genotyping and prognostic value of HPV E4 protein and transcription factors NANOG and SOX11 in atypical p16 patchy squamous epithelium of cervix

Maja Kebe Radulovic

Maja Kebe Radulovic

Institute of Pathology, Faculty of Medicine, University of LjubljanaLjubljana, Slovenia
Search for this author on
Sciendo|Google Scholar
Radulovic, Maja Kebe
, 
Anja Ostrbenk

Anja Ostrbenk

Institute of Microbiology and Immunology, Faculty of Medicine, University of LjubljanaLjubljana, Slovenia
Search for this author on
Sciendo|Google Scholar
Ostrbenk, Anja
, 
Mario Poljak

Mario Poljak

Institute of Microbiology and Immunology, Faculty of Medicine, University of LjubljanaLjubljana, Slovenia
Search for this author on
Sciendo|Google Scholar
Poljak, Mario
 and 
Margareta Strojan-Flezar

Margareta Strojan-Flezar

Institute of Pathology, Faculty of Medicine, University of LjubljanaLjubljana, Slovenia
Search for this author on
Sciendo|Google Scholar
Strojan-Flezar, Margareta
  
Jul 08, 2025
Human papillomavirus (HPV) genotyping and prognostic value of HPV E4 protein and transcription factors NANOG and SOX11 in atypical p16 patchy squamous epithelium of cervix's Cover Image
About this article
Previous Article
Next Article
Cite
Download Cover
Article Category: Research Article
Published Online: Jul 08, 2025
Page range: 391 - 402
Received: Mar 16, 2025
DOI: https://doi.org/10.2478/raon-2025-0038
Keywords
© 2025 Maja Kebe Radulovic et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1.

Images of atypical squamous epithelium cases with patchy p16 reaction: (A–F). The same sample stained with HE and p16 at 200× magnification. Panels (A), (C), and (E) show HE staining, while panels (B), (D), and (F) show corresponding p16 staining.
Images of atypical squamous epithelium cases with patchy p16 reaction: (A–F). The same sample stained with HE and p16 at 200× magnification. Panels (A), (C), and (E) show HE staining, while panels (B), (D), and (F) show corresponding p16 staining.

Figure 2.

Only case in the group with atypical squamous epithelium with patchy p16 expression and positive reaction for E4 human papillomavirus (HPV). The tissue was positive for HPV16 and 31 and the follow-up was negative. The same section at 100x magnification: (A) HE staining; (B) p16 staining, (C) Ki67 staining, (D) E4 HPV staining, (E) Nanog staining, F. SOX11 staining.
Only case in the group with atypical squamous epithelium with patchy p16 expression and positive reaction for E4 human papillomavirus (HPV). The tissue was positive for HPV16 and 31 and the follow-up was negative. The same section at 100x magnification: (A) HE staining; (B) p16 staining, (C) Ki67 staining, (D) E4 HPV staining, (E) Nanog staining, F. SOX11 staining.

Figure 3.

One of the three cases in the group with atypical squamous epithelium with patchy p16 with CIN3 on follow-up. The tissue was positive for human papillomavirus (HPV) 52, 53,56, 66, 73. The same section at 100x magnification: (A) HE. (B) p16 staining. (C) Ki67 staining. (D) E4 HPV staining. (E) Nanog staining. (F) SOX11 staining.
One of the three cases in the group with atypical squamous epithelium with patchy p16 with CIN3 on follow-up. The tissue was positive for human papillomavirus (HPV) 52, 53,56, 66, 73. The same section at 100x magnification: (A) HE. (B) p16 staining. (C) Ki67 staining. (D) E4 HPV staining. (E) Nanog staining. (F) SOX11 staining.

Figure 4.

Immunohistochemical reactions in the only cervical intraepithelial neoplasia 3 (CIN3) case with a positive SOX11 reaction in study. The same section at 100x magnification: (A) HE. (B) p16 staining. (C) Ki67 staining. (D) E4 HPV staining. (E) Nanog staining. (F) SOX11 staining.
Immunohistochemical reactions in the only cervical intraepithelial neoplasia 3 (CIN3) case with a positive SOX11 reaction in study. The same section at 100x magnification: (A) HE. (B) p16 staining. (C) Ki67 staining. (D) E4 HPV staining. (E) Nanog staining. (F) SOX11 staining.

Biomarkers in the primary biopsy and follow-up in the groups with atypical squamous epithelium with a patchy and a null reaction to p16

Group Case Ki67 E4 HPV Nanog HPV genotypes Follow-up sample Follow-up diagnosis Time from primary biopsy to follow-up sample
p16 1 1/3 - 1+ 31, 52 biopsy CIN1 2 years
null 2 3/3 - - 53 biopsy CIN2 2 years
3 1/3 - - - biopsy CIN3 4 months
p16 patchy 4 0 - - 39 cone CIN1 8 months
5 1/3 - - - cone CIN1 6 years
6 3/3 - 1+ 53 cone CIN3 5 years
7 3/3 - - 52, 53, 56, 66, 73 Cervical amputation CIN3 10 months
8 1/3 - - 44 cone CIN3 10 months

Comparison of different biological markers between groups using Spearman correlation

Groups p16 Ki67 HPV E4 Nanog lrHPV tissue hrHPV tissue hrHPV: tissue+smear lrHPV: tissue+smear
p16 null vs. p16 patchy r = 1 p = 0.000 r = 0.000 r = 0.116 r = 0.089 r = 0.209 r = 0.384 p = 0.025 r = 0.274 r = 0.285
p16 patchy vs. condyloma r = 0.000 r = 0.535 p = 0.001 r =0.280 r = 0.494 p = 0.004 r = 0.458 p = 0.007 r = 0.000 r=-0.186 r = 0.494 p = 0.004
p16 patchy vs. CIN1 r = 0343 p = 0.041 r = 0.35 p = 0.037 r = 0.81 p = 0.000 r = 0.532 p = 0.001 r = 0.354 p = 0.034 r = 0.161 r = -0.039 r = 0.359 p = 0.032
p16 patchy vs. CIN2 r = 1 p = 0.000 r = 0.66 p = 0.000 r = 0.728 p = 0.000 r = 0.632 p = 0.000 r = -0.193 r = 0.45 p = 0.007 r = 0.176 r = -0.089
p16 patchy vs. CIN3 r = 1 p = 0.000 r = 0.683 p = 0.000 r = 0.280 r = 0.575 p = 0.000 r = -0.375 p = 0.032 r = 0.594 p = 0.000 r = 0.433 p = 0.012 r = -0.433 p = 0.012
p16 null vs. condyloma r = 1 r = 0.617 p = 0.004 r = 0.369 r = 0.664 p = 0.001 r = 0.704 p = 0.001 r = 0.464 p = 0.039 r = 0.066 r = 0.818 p = 0.000
p16 null vs. CIN1 r = 0.963 p = 0.000 r = 0.394 r = 0.84 p = 0.000 r = 0.691 p = 0.000 r = 0.568 p = 0.005 r = 0.569 p = 0.005 r = 0.233 r = 0.652 p = 0.001
p16 null vs. CIN2 r = 1 r = 0.735 p = 0.000 r = 0.756 p = 0.000 r = 0.728 p = 0.000 r = 0.000 r = 0.832 p = 0.000 r = 0.455 p = 0.034 r = 0.204
p16 null vs. CIN3 r = 1 r = 0.783 p = 0.000 r = 0.369 r = 0.665 p = 0.001 r = -0.302 r = 1 r = 0.739 p = 0.000 r = -0.302
Atypia (p16 patchy and null) vs. CIN2 r = 0.811 p = 0.000 r = 0.607 p = 0.000 r = 0.751 p = 0.000 r = 0.598 p = 0.000 r = -0.125 r = 0.521 p = 0.000 r = 0.231 r = -0.007

Immunohistochemical reactions to various biological markers and human papillomavirus (HPV) genotyping in tissue biopsy and previous cervical smear, where available, by study group

Group p16 N Ki67 N E4 HPV N Nanog N HPV genotyping N tissue N smear
p16 null N = 11 Null 11 0 5 Neg. 11 Neg. 7 Neg 9 3
Patchy 0 1/3 4 Pos. 0 1 4 lrHPV only 2 1
En Block 0 2/3 0 2 0 hrHPV 0 3
3/3 2 3 0
p16 patchy N = 24 Null 0 0 12 Neg. 23 Neg. 13 Neg 10 2
Patchy 24 1/3 5 Pos. 1 1 11 lrHPV only 6 2
En Block 0 2/3 3 2 0 hrHPV 8 12
3/3 4 3 0
Condyloma N = 9 Null 0 0 0 Neg. 7 Neg. 0 Neg 0 1
Patchy 9 1/3 2 Pos. 2 1 9 lrHPV only 6 2
En Block 0 2/3 1 2 0 hrHPV 3 1
3/3 6 3 0
CIN1 N = 12 Null 0 0 2 Neg. 2 Neg. 0 Neg 0 0
Patchy 10 1/3 2 Pos. 10 1 12 lrHPV only 6 1
En Block 2 2/3 4 2 0 hrHPV 6 3
3/3 4 3 0
CIN2 N = 11 Null 0 0 0 Neg. 3 Neg. 0 Neg 1 0
Patchy 0 1/3 0 Pos. 8 1 7 lrHPV only 2 0
En Block 11 2/3 2 2 4 hrHPV 8 3
3/3 9 3 0
CIN3 N = 9 Null 0 0 0 Neg. 7 Neg. 1 Neg 0 0
Patchy 0 1/3 0 Pos. 2 1 3 lrHPV only 0 0
En Block 9 2/3 0 2 3 hrHPV 9 4
3/3 9 3 2

j_raon-2025-0038_Utab_001

1
0,9
0,8
0,7
0,6
0,5
0,4
0,3
0,2
0,1
0
-0,1
-0,2
-0,3
-0,4
-0,5
-0,6
-0,7
-0,8
-0,9
-1

Follow-up histology, cytology, human papillomavirus (HPV) result by study groups (p16 null and p16 patchy)

Group Follow up histo N Follow up cyto N Follow up HPV N
p16 null (11 cases, 9 biopsies) Neg 0 Neg 5 Neg 5
CIN1 1 ASC-US 4 Pos 3
CIN2 1 LSIL 0
CIN3 1 HSIL 0
p16 patchy (24 cases, 17 biopsies) Neg 0 Neg 5 Neg 4
CIN1 2 ASC-US 6 Pos 5
CIN2 0 LSIL 4
CIN3 3 HSIL 2
Language:
English
Publication timeframe:
4 times per year
Journal Subjects:
Medicine, Clinical Medicine, Internal Medicine, Haematology, Oncology, Radiology
Journal RSS Feed