ISSN 2415-3060 (print), ISSN 2522-4972 (online)
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УЖМБС 2020, 5(1): 72–78
Experimental Medicine and Morphology

Immunohistochemical Characteristics of Proliferative and Apoptotic Processes in Testicular Embryonal Carcinoma

Potapov S. M., Galata D. I., Pliten O. M.

Rising of the incidence rate of testicular germ cell tumors in combination with the predominantly young age of patients gives this problem medical and social significance. The purpose of the work was to establish proliferative and apoptotic processes in testicular embryonal carcinoma. Material and methods. The study was performed on 13 surgically removed testicles affected by embryonal carcinoma. Tumors were grouped according to the degree of tumorous progression in accordance with the pTNM classification. For evaluation of proliferative and apoptotic processes the relative area (S) and intensity (L) of Ki–67, Bax, Bcl–2, and p53 expression were studied. The proliferation index was calculated as well. Results and discussion. The study determined increasing of average S, L values of Ki–67 expression and proliferation index. We also revealed very high and high positive correlation between proliferation index and S with L of Ki–67 expression (r=+0.94; r=+0.83; p<0.05, respectively) during the transition from the initial to the late stages of tumorous progression. S of Bax expression was also increasing at the transition from early to late stages of tumorous progression, and L expression remained low regardless of stage. High positive correlation between S of Bax and S of Ki–67 expression and between S of Bax and proliferation index (r=+0.84; r=+0.82; p<0.05, respectively) was revealed during the transition from the initial to the late stages of tumorous progression. In patients with distant metastases and metastases to the lymph nodes S of Bax expression was bigger than that in patients without metastases. The activity of anti–apoptotic protein bcl–2 in the embryonal carcinoma was insignificant and determined predominantly in the cells of immune infiltrate. A very high and high negative correlation was found between S of bcl–2 expression and S of Ki–67 expression with proliferation index (r= –0.92; r=–0.88; p<0.05, respectively) during the transition from the initial to the late stages of tumorous progression. Correlation analysis revealed a high positive relationship between S of p53 expression and S of Bax and Ki–67 expression (r=+0.82; r=+0.71; p<0.05, respectively) during the tumorous progression and a moderate positive relationship between S of p53 expression and proliferation index (r=+0.69; p<0.05). In patients with lymphogenic metastases S of p–53 expression was bigger than that in patients without metastases (p<0.05). Conclusion. Thus, the use of Ki–67, Bax, bcl–2, and p53 markers in the diagnosis of testicular tumors is advisable, since they reflect the malignant potential of the tumor, and can be used as prognostic markers.

Keywords: embryonal carcinoma, proliferation, apoptosis, immunohistochemistry

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  1. Burova EA, Bulanov AA, Tryakin AA, Fedyanin MYu, Tyulyandin SA, Matveev VB. Lechenie seminomy yaichka I stadii [Treatment for stage I testicular seminoma]. Cancer Urology. 2010; 3: 7–11 [Ukrainian]
  2. Eble JN, Sauter G, Epstein JI, Sesterhenn IA, editors. World Health Organization Classification of Tumours. Pathology and Genetics of Tumours of the Urinary System and Male Genital Organs. Lyon: IARC Press; 2004. 359 p.
  3. Trabert B, Chen J, Devesa SS, Bray F, McGlynn KA. International patterns and trends in testicular cancer incidence, overall and by histologic subtype, 1973–2007. Andrology. 2015 Jan; 3(1): 4–12. PMID: 25331326; PMCID: PMC4410839.
  4. Fedorenko ZP, Michailovich YY, Goulak LO, Gorokh YeL, Ryzhov AYu, et al. Bulletin of national cancer registry of Ukraine 2017–2018. Kyiv. 2019; 20. [Internet]. Available from:
  5. Moch H, Humphrey PA, Ulbright ThM, Reuter VE, Eds. WHO Сlassification of Tumours of the Urinary System and Male Genital Organs. Lyons: IARC Press; 2016. 356 р.
  6. Lobo J, Costa AL, Vilela–Salgueiro B, Rodrigues Â, Guimarães R, Cantante M, et al. Testicular germ cell tumors: revisiting a series in light of the new WHO classification and AJCC staging systems, focusing on challenges for pathologists. Hum Pathol. 2018 Dec; 82:113–124. PMID: 30041024.
  7. Patent 119922 Ukraine. Sposib kilkisnoi otsinky rivnia svitlosti ta vidnosnoi ploshchi ekspresii markeriv pry imunohistokhimichnomu doslidzhenni tkanyn / Potapov SM, Markovskiy VD, KulIshova NE (UA); zayavnik i vlasnik patentu Kharkivskyi natsionalnyi medychnyi universytet (UA); zayavl 2019 Serp 27. [Ukrainian]
  8. Kobzar AI. Prikladnaya matematicheskaya statistika. Dlya inzhenerov i nauchnyih rabotnikov. [Internet]. FIZMATLIT, 2012. 816 p. Available from: [Russian]
  9. Runyon RP. Nonparametric Statistics: A Contemporary Approach (Addison–Wesley series in statistics). Addison–Wesley Publishing Co; 1977. 218 p.
  10. Shatseva TA, Muhina MS. Antigen Ki–67 v otsenke opuholevoy proliferatsii. Ego struktura i funktsii [Ki–67 antigen in the evaluation of tumor proliferation. Its structure and functions]. Voprosy onkologii. 2004; 2(50): 157–63. [Russian]
  11. Yurin AG, Kovalskiy GB. Issledovanie proliferativnoy aktivnosti po ekspressii antigena Ki–67 v odinochnyih i mnozhestvennyih sinhronnyih epitelialnyih opuholyah tolstoy kishki [Proliferative activity by expression of antigen Ki–67 in solitary and multiple synchroneous epithelial tumors of the colon]. Arhiv patologii. 2005; 5 (67): 38–41. [Russian]
  12. Pozharisskiy KM, Leenman EE. Znachenie immunogistohimicheskih metodik dlya opredeleniya haraktera lecheniya i prognoza opuholevyih zabolevaniy [Role of immunohistochemical techniques in planning treatment and prognosis of tumors]. Arhiv patologii. 2000; 5 (62): 3–11. [Russian]
  13. Walensky LD, Gavathiotis E. BAX unleashed: the biochemical transformation of an inactive cytosolic monomer into a toxic mitochondrial pore. Trends Biochem Sci. 2011 Dec; 36(12): 642–52. PMID: 21978892; PMCID: PMC3454508.
  14. Fauvet R, Dufournet C, Poncelet C, Uzan C, Hugol D, Daraï E. Expression of pro–apoptotic (p53, p21, bax, bak and fas) and anti–apoptotic (bcl–2 and bcl–x) proteins in serous versus mucinous borderline ovarian tumours. J Surg Oncol. 2005 Dec 15; 92(4): 337–43. PMID: 16299808
  15. Bose P, Klimowicz AC, Kornaga E, Petrillo SK, Matthews TW, Chandarana S, et al. Bax expression measured by AQUAnalysis is an independent prognostic marker in oral squamous cell carcinoma. BMC Cancer. 2012 Aug 1; 12: 332. PMID: 22852863; PMCID: PMC3487960.–2407–12–332
  16. Dawson SJ, Makretsov N, Blows FM, Driver KE, Provenzano E, Le Quesne J, et al. BCL2 in breast cancer: a favourable prognostic marker across molecular subtypes and independent of adjuvant therapy received. Br J Cancer. 2010 Aug 24; 103(5): 668–75. PMID: 20664598. PMCID: PMC2938244.
  17. Grange F, Petrella T, Beylot–Barry M, Joly P, D'Incan M, Delaunay M, et al. Bcl–2 protein expression is the strongest independent prognostic factor of survival in primary cutaneous large B–cell lymphomas. Blood. 2004 May 15; 103(10): 3662–8. PMID: 14726400
  18. Pieretti M, Hopenhayn–Rich C, Khattar NH, Cao Y, Huang B, Tucker TC. Heterogeneity of ovarian cancer: relationships among histological group, stage of disease, tumor markers, patient characteristics, and survival. Cancer Invest. 2002; 20(1): 11–23. PMID: 11852993
  19. Zolota V, Tsamandas AC, Melachrinou M, Batistatou A, Scopa C. Expression of CD44 protein in renal cell carcinomas: association with p53 expression. Urol Oncol. 2002 Jan–Feb; 7(1): 13–7. PMID: 12474536
  20. Jones RH, Vasey PA. New directions in testicular cancer; molecular determinants of oncogenesis and treatment success. Eur J Cancer. 2003 Jan; 39(2): 147–56. PMID: 12509945