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Cytotoxic Effects of the Selective Ligands of Membrane Progesterone Receptors in Human Pancreatic Adenocarcinoma Cells BxPC3

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Abstract

Progesterone and its synthetic analogues act on cells through different types of receptors, affecting proliferation and apoptosis. These compounds exert their effect through the nuclear receptors and the insufficiently studied membrane progesterone receptors (mPRs) belonging to the progestin and adiponectin Q receptor (PAQR) family. We have identified two selective ligands of mPRs that activate only this type of progesterone receptors – 19-hydroxypregn-4-en-20-one (LS-01) and 19-hydroxy-5β-pregn-3-en-20-one (LS-02). The goal of this work is to study the effect of these compounds on proliferation and death of human pancreatic adenocarcinoma cells BxPC3 and involvement of the two kinases (p38 MAPK and JNK) in signaling pathways activated by progestins through mPRs. It was shown that progesterone and the compound LS-01 significantly (p < 0.05) inhibited the BxPC3 cell viability, with JNK serving as a mediator. The identified targets of these two steroids are the genes of the proteins Ki67, cyclin D1, PCNA, and p21. Progesterone and the compound LS-01 significantly (p < 0.05) stimulate DNA fragmentation, enhancing the cell death. The p38 mitogen-activated protein kinase (MAPK) is a key mediator of this process. The BCL2A1 protein gene was identified as a target of both steroids. The compound LS-02 significantly (p < 0.05) alters membrane permeability and changes the exposure of phosphatidylserine on the outer membrane leaflet, also enhancing the cell death. This compound acts on these processes by activating both kinases, JNK and p38 MAPK. The compound LS-02 targets the genes encoding the proteins HRK, caspase 9, and DAPK.

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Abbreviations

BCL2A1:

BCL2 related protein A1

cyclin D1:

type D1 cyclin-dependent kinases activator

DAPK1:

death-associated protein kinase 1

FAS:

Fas cell surface death receptor

HRK:

hara-kiri

Ki67:

marker of proliferation

mPRs:

membrane progesterone receptors

nPRs:

nuclear progesterone receptors

P4:

progesterone

p21:

cyclin-dependent kinase inhibitor 1A

p27:

a cyclin-dependent kinase inhibitor 1B

PCNA:

proliferating cell nuclear antigen

qPCR:

quantitative real-time PCR

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Acknowledgments

The authors express their gratitude to the Center of Collective Use “Genome” of the Engelhard Institute of Molecular Biology (Russian Academy of Sciences) and to I. Yu. Petrushanko for helping with the flow cytometry experiments.

Funding

This work was financially supported by the Russian Foundation for Basic Research (project no. 20-015-00092).

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Authors and Affiliations

  1. Faculty of Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    Alexey I. Goncharov, Viktoriia L. Shliapina, Olga V. Smirnova & Tatiana A. Shchelkunova

  2. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 119991, Moscow, Russia

    Inna S. Levina & Igor V. Zavarzin

  3. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991, Moscow, Russia

    Ivan A. Morozov & Petr M. Rubtsov

Authors
  1. Alexey I. Goncharov
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  2. Inna S. Levina
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  3. Viktoriia L. Shliapina
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Correspondence to Tatiana A. Shchelkunova.

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The authors declare no conflicts of interest in financial or any other sphere. This article does not contain any studies with human participants or animals performed by any of the authors.

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Goncharov, A.I., Levina, I.S., Shliapina, V.L. et al. Cytotoxic Effects of the Selective Ligands of Membrane Progesterone Receptors in Human Pancreatic Adenocarcinoma Cells BxPC3. Biochemistry Moscow 86, 1446–1460 (2021). https://doi.org/10.1134/S0006297921110080

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