Exploration of protein PCP4 as a potential tumor marker in uterine leiomyoma
Kuznetsova M.V., Shevelev A.B., Pozdnyakova N.V., Samoilova D.V., Vypryazhkina V.E., Tonoyan N.M., Trubnikova E.V., Zelensky D.V., Vishnyakova P.A.
Objective: Evaluation of the possible role of Purkinje cell protein 4 (PCP4) as a potential tumor marker of uterine leiomyoma by measurement of antibody titer against this protein in blood serum and the applicability of this technique for evaluation of proteins as potential vaccine antigens.
Materials and methods: cDNA fragment clone was derived from PCP4 gene in leiomyoma nodule. After that the soluble PCP4-GFP chimera based on E. coli was constructed, and the recombinant protein was purified. This product was used to evaluate the upper limit of titer to detect antibodies in blood serum in patients using indirect enzyme immunoassay. Serum samples (24) were collected from donors among the patients undergoing treatment at the Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, who gave informed consent to participate in the study. Four groups of patients were formed to study the immunological activity of sera against the 6His-PCP4hs-GFP protein. Group 1 was the control group and was composed of men. Group 2 consisted of women without diagnosed fibroids who had five or more successful pregnancies in history. Group 3 included women with fibroids, who also had five or more successful pregnancies in history. Group 4 included women with recurrent myomas.
Results: The pRSET-EmGFP expression vector containing the PCP4 gene and the reporter gene of the green fluorescent protein EmGFP was derived from the cDNA of the myoma nodule with a driver mutation in the MED12 gene. Human PCP4 in preparative amounts was obtained using this construct embedded in the E. coli genome, that was sufficient for analysis of its reaction with antibodies from blood serum in different groups of patients. Blood sera in the control group of men showed high immunological reactivity against the 6His-PCP4hs-GFP protein, whereas the difference between the groups of women was insignificant. Minimum difference between sera was found in groups 2 and 4. Moreover, the reaction of sera in women with recurrent fibroids was higher compared with women without fibroids, who had five or more successful pregnancies. Comparison of women in groups 3 and 4 showed statistically significant differences in the optical density values obtained in dilution of sera 1:1600, 1:3200 and 1:6400. At the same time the reaction of sera of women with recurrent fibroids was higher compared with sera of women with fibroids, who had five or more successful pregnancies.
Based on the level of activity, sera were divided into 3 categories. It was found that reaction of sera of men against the 6His-PCP4hs-GFP antigen was most pronounced. The proportion of high activity in sera was 60%. In the group of women without fibroids, who had five or more successful pregnancies, the proportion of low activity in sera was 44%, that is the maximum value of indicator compared with the other groups. The group of women with fibroids, who had five or more successful pregnancies, is characterized by the moderate level of reactivity with the 6His-PCP4hs-GFP antigen. This category accounts for 75% of sera, that is the highest value of indicator among all groups. Finally, the group of women with recurrent fibroids is characterized by equal number of sera with high and moderate activity, that shows a greater tendency for production of antibodies against PCP4/PEP19 in this group of patients compared with multiparous women.
Conclusion: For the first time, human PCP4 was in preparative amounts sufficient for analysis of its reaction with antibodies from blood serum in different groups of patients. Testing showed that most samples in all groups had significant titers of antibodies against PCP4. The activity of these antibodies varies widely both in sera of men and women. Antibodies against PCP4 are most common in women with recurrent fibroids compared with multiparous women, especially without leiomyomas. Thus, it is unlikely that PCP4 immunization сarries the risk of any pathology. The titers of antibodies to PCP4 in the general group of multiparous women were significantly lower compared with men and patients with recurrent fibroids. Therefore, using PCP4 as the basis to develop a preventive vaccine against leiomyomas is not advisable.
Authors' contributions: Kuznetsova M.V. – literature review, development of the concept and design of the study; Pozdnyzkova N.V. – protein cloning; Vypryazhkina V.E. – identification of proteins, blot analysis; Trubnikova E.V., Zelensky D.V. – statistical data processing; Kuznetsova M.V., Shevelev A.B. – text writing; Vishnyakova P.А. – final text editing.
Conflicts of interest: The authors confirm that they have no conflicts of interest to declare.
Funding: The study was conducted within the RNF project 23-15-00069 "Prophilactic vaccine development to prevent uterine leiomyoma in patients planning pregnancy".
Ethical Approval: The study was approved by the Ethics Committee of the Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia
Patient Consent for Publication: The patients have signed informed consent for participation in the study and publication of the obtained data.
Authors' Data Sharing Statement: The data supporting the findings of this study are available on request from the corresponding author after approval from the principal investigator.
For citation: Kuznetsova M.V., Shevelev A.B., Pozdnyakova N.V., Samoilova D.V., Vypryazhkina V.E.,
Tonoyan N.M., Trubnikova E.V., Zelensky D.V., Vishnyakova P.A. Exploration of protein PCP4 as
a potential tumor marker in uterine leiomyoma.
Akusherstvo i Ginekologiya/Obstetrics and Gynecology. 2025; (8): 159-171 (in Russian)
https://dx.doi.org/10.18565/aig.2025.171
Keywords
uterine fibroids
gene expression
PCP 4
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Received 26.06.2025
Accepted 11.08.2025
About the Authors
Maria V. Kuznetsova, PhD, Senior Researcher at the Institute of Reproductive Genetics, V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Oparin str., 4, +7(495)438-13-41, mkarja@mail.ru, https://orcid.org/0000-0003-3790-0427Alexey B. Shevelev, Chief Researcher, N.I. Vavilov Institute of General Genetics of the Russian Academy of Sciences, 119991, Russia, GSP-1, Moscow, Gubkin str., 3, +7(915)087-52-93, shevel_a@hotmail.com, https://orcid.org/0000-0003-3564-7405
Natalia V. Pozdnyakova, PhD, Senior Researcher at the Laboratory of Radionuclide and Radiation Technologies in Experimental Oncology, Blokhin National Research Medical Center of Oncology, Ministry of Health of Russia, 115230, Russia, Moscow, Kashirskoe Shosse, 23, +7(977)767-99-90, natpo2002@mail.ru,
https://orcid.org/0000-0002-5765-3016
Darya V. Samoilova, Researcher at the Central Pathoanatomical Laboratory of the Academician A.P. Avtsyn Research Institute of Human Morphology, Russian Scientific Center of Surgery named after Academician B.V. Petrovsky, 109548, Russia, Moscow, Ugreshskaya str., 2-7, +7(916)594-89-06, dashasam@mail.ru,
https://orcid.org/0000-0001-5639-0835
Viktoria E. Vypryazhkina, PhD, Senior Researcher at the Laboratory of Molecular Genetic Methods, Institute of Translation Medicine, V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Oparin str., 4, +7(495)438-13-41.
Narine M. Tonoyan, PhD, Doctor, V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Oparin str., 4, SPIN: 8547-9399, Scopus AuthorID: 57213609878, https://orcid.org/0000-0002-1631-1829
Elena V. Trubnikova, Dr. Bio. Sci., Associate Professor, Chief Researcher at the Laboratory of Genetics, Kursk State University, 305000, Russia, Kursk, Radishchev str., 33; Leading Researcher, N.I. Vavilov Institute of General Genetics of the Russian Academy of Sciences, 119991, Russia, GSP-1, Moscow, Gubkin str., 3, +7(910)311-18-86,
tr_e@list.ru, https://orcid.org/0000-0001-5025-9406
Dmitry V. Zelensky, PhD student, Kursk State University, 305000, Russia, Kursk, Radishchev str., 33, +7(999)700-38-81, dmitriizelenskii@mail.ru,
https://orcid.org/0000-0006-0490-7760
Polina A. Vishnyakova, PhD, Head of the Laboratory of Regenerative Medicine, V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Oparin str., 4; Head of the Laboratory of Molecular Pathophysiology, Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples’ Friendship University of Russia, Scopus AuthorID: 57190971385, https://orcid.org/0000-0001-8650-8240
Corresponding author: Maria V. Kuznetsova, mkarja@mail.ru
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