PC Kz is the novel human prostate cancer model in vitro and in vivo

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Abstract

BACKGROUND: The use of relevant in vitro and in vivo model systems is important in preclinical studies of anticancer agents. The process of creating tumor models is methodically complicated and has a number of disadvantages. Among tumor models of prostate cancer, the most accessible are 2D models (DU145, 22Rv1, PC3, LNCaP, VCaP cell lines), their xenograft models in immunodeficient mice and some patient-derived xenograft models. However, this panel of experimental models is not perfect and needs further expansion.

AIM: To create a new preclinical prostate cancer model, characterize it (morphology, tumorigenicity, tumor growth kinetics in vivo, verification of prostate-specific membrane antigen expression status, testosterone production, sensitivity to CYP17A1 inhibitors), and develop resistance to the steroid CYP17A1 inhibitor — abiraterone.

METHODS: CYP17A1 expression in PC Kz cell line was evaluated by reverse transcription polymerase chain reaction. Testosterone concentration was determined with an enzyme-linked immunosorbent assay. The sensitivity to antitumor agents was studied with the MTT test. Tumorigenicity was evaluated by transplantation of PC Kz cell line into Balb/c nude mice. The prostate-specific membrane antigen expression status was assessed using the indirect reaction of surface immunofluorescence.

RESULTS: The PC Kz cell line is characterized by a high level of CYP17A1 messenger RNA expression, comparable to that of the commercial 22Rv1 cell line. Immunophenotypic analysis demonstrated negative prostate-specific membrane antigen expression status of PC Kz cell line. A significant decrease (18%) in testosterone concentration in vitro was found, compared to the value in the control. This effect can be associated with the suppression of CYP17A1 gene expression.

The studied PC Kz cell line is tumorigenic in Balb/c nude mice (100% tumorigenicity was detected during the first passage at a transplantation dose of 107 cells/mouse). A pathomorphological study of the structures of the obtained subcutaneous PC Kz xenografts verified their identity to the histological image of human prostate cancer. Furthermore, PC Kz/AA cell line was obtained, resistant to abiraterone; the index of resistance was 3.4.

CONCLUSION: The derived PC Kz cell line was adapted for in vitro and in vivo growth, characterized by the main biological parameters, and can be recommended as an adequate test system to use in preclinical studies of new antitumor agents for the human prostate cancer treatment.

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About the authors

Darina V. Sokolova

N.N. Blokhin National Medical Research Center of Oncology; Peoples’ Friendship University of Russia; Sirius University of Science and Technology

Email: v.pokrovsky@ronc.ru
ORCID iD: 0000-0003-3972-2425
SPIN-code: 2960-4800

Cand.  Sci. (Bio.)

Russian Federation, 24 Kashirskoe shosse, Moscow 115478; Moscow; Sochi

Irina I. Khan

N.N. Blokhin National Medical Research Center of Oncology; Peoples’ Friendship University of Russia

Email: irinchek05@gmail.com
ORCID iD: 0000-0003-2948-0872
SPIN-code: 6826-7694

Cand.  Sci. (Bio.)

Russian Federation, 24 Kashirskoe shosse, Moscow 115478;Moscow

Dmitry D. Zhdanov

Institute of Biomedical Chemistry; Peoples’ Friendship University of Russia

Email: zhdanovdd@mail.ru
ORCID iD: 0000-0003-4753-7588
SPIN-code: 3845-2544

Dr. Sci. (Bio.)

Russian Federation, Moscow; Moscow

Elena A. Demidova

N.N. Blokhin National Medical Research Center of Oncology

Email: badjito@mail.ru
ORCID iD: 0000-0002-6511-3423
Russian Federation, 24 Kashirskoe shosse, Moscow 115478

Valery O. Krivchenko

Moscow Institute of Physics and Technology

Email: krivchenko.vo@phystech.edu
ORCID iD: 0009-0000-0807-1418
SPIN-code: 6971-4087
Russian Federation, Moscow

Chingis Aidossov

Peoples’ Friendship University of Russia

Email: aidossovchina@gmail.com
ORCID iD: 0009-0001-0034-8731
Russian Federation, Moscow

Vadim S. Pokrovsky

N.N. Blokhin National Medical Research Center of Oncology; Peoples’ Friendship University of Russia; Sirius University of Science and Technology

Author for correspondence.
Email: v.pokrovsky@ronc.ru
ORCID iD: 0000-0003-4006-9320
SPIN-code: 4552-1226

MD, Dr. Sci. (Med.), Assistant Professor

Russian Federation, 24 Kashirskoe shosse, Moscow 115478; Moscow; Sochi

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Supplementary files

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2. Fig. 1. Structures of the title compounds: a — alsevirone, 2’-[((E)-3β-hydroxyandrost-5-en-17-ylidene)methyl]-4’,5’-dihydro-1’,3’-oxazole; b — abiraterone— 3-(3β-hydroxyandrosta-5,16-dien-17-yl)-pyridine.

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3. Fig. 2. Prostate cancer cell line PC Kz: a — cells adhesion obtained by mechanical and enzymatic methods of disaggregation of the resected tissue, magnification ×50; b — general view of the explant with tumor cells migrating from it, magnification ×50; c — radial migration of tumor cells from the explant, magnification ×100; d — cell morphology, magnification ×200; e — PC Kz cell line fibroblasts contamination, magnification ×200.

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4. Fig. 3. Duration of the passages (from the 1st to 10th) to 100% confluency of PC Kz cell line.

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5. Fig. 4. Balb/c nude mouse with PC Kz xenograft (1st passage, 42nd day after cell line transplantation).

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6. Fig. 5. Growth dynamics of the 1st passage subcutaneous xenografts of PC Kz, 22Rv1 and DU145 in Balb/c nude mice.

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7. Fig. 6. Histological image of a subcutaneous PC Kz xenograft in Balb/c nude mouse, stained with hematoxylin: a — cell detritus in the tumor parenchyma, ×400; b — thickened part of the connective tissue capsule with blood vessels, ×200; c — general morphological picture of the tumor parenchyma: 1 — accumulation of macrophages, 2 — blood vessel, 3 — mitotically active cell, 4 — fibroblast (×400); d — formed nested structures (circled), ×400.

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8. Fig. 7. Normalized to 18S gene CYP17A1 expression in PC Kz cells line in comparison with commercial prostate cancer cell lines.

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9. Fig. 8. Immunophenotypic analysis of PC Kz cell line; along the vertical axis — cell count; along the horizontal axis — fluorescence intensity. The marker was set according to isotype control.

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10. Fig. 9. Duration of passages to 100% confluence after adding abiraterone to the culture medium of PC Kz cell line.

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11. Fig. 10. Effect of tested drugs on the proliferative activity of native PC Kz and PC Kz/AA resistant to abiraterone cell line: a — abiraterone; b — alsevirone; c — paclitaxel (Taxol®); d — cisplatin; e — doxorubicin.

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