Effects of Progesterone and Selective Ligands of Membrane Progesterone Receptors in HepG2 Cells of Human Hepatocellular Carcinoma

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Tatiana A. Shchelkunova^1,a*, Inna S. Levina², Ivan A. Morozov³, Petr M. Rubtsov³, Alexey I. Goncharov¹, Yury V. Kuznetsov², Igor V. Zavarzin², and Olga V. Smirnova¹

¹Faculty of Biology, Lomonosov Moscow State University, 119234 Moscow, Russia

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

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

^* To whom correspondence should be addressed.

Received December 2, 2022; Revised September 13, 2023; Accepted September 22, 2023

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Progesterone exerts multiple effects in different tissues through nuclear receptors (nPRs) and through membrane receptors (mPRs) of adiponectin and progestin receptor families. The effect of progesterone on the cells through different types of receptors can vary significantly. At the same time, it affects the processes of proliferation and apoptosis in normal and tumor tissues in a dual way, stimulating proliferation and carcinogenesis in some tissues, suppressing them and stimulating cell death in others. In this study, we have shown the presence of high level of mPRβ mRNA and protein in the HepG2 cells of human hepatocellular carcinoma. Expression of other membrane and classical nuclear receptors was not detected. It could imply that mPRβ has an important function in the HepG2 cells. The main goal of the work was to study functions of this protein and mechanisms of its action in human hepatocellular carcinoma cells. Previously, we have identified selective mPRs ligands, compounds LS-01 and LS-02, which do not interact with nuclear receptors. Their employment allows differentiating the effects of progestins mediated by different types of receptors. Effects of progesterone, LS-01, and LS-02 on proliferation and death of HepG2 cells were studied in this work, as well as activating phosphorylation of two kinases, p38 MAPK and JNK, under the action of three steroids. It was shown that all three progestins after 72 h of incubation with the cells suppressed their viability and stimulated appearance of phosphatidylserine on the outer surface of the membranes, which was detected by binding of annexin V, but they did not affect DNA fragmentation of the cell nuclei. Progesterone significantly reduced expression of the proliferation marker genes and stimulated expression of the p21 protein gene, but had a suppressive effect on the expression of some proapoptotic factor genes. All three steroids activated JNK in these cells, but had no effect on the p38 MAPK activity. The effects of progesterone and selective mPRs ligands in HepG2 cells were the same in terms of suppression of proliferation and stimulation of apoptotic changes in outer membranes, therefore, they were mediated through interaction with mPRβ. JNK is a member of the signaling cascade activated in these cells by the studied steroids.

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KEY WORDS: progesterone, selective ligands of membrane progesterone receptors, proliferation, apoptosis, nuclear progesterone receptor, membrane progesterone receptor β, human HepG2 hepatocellular carcinoma cells

DOI: 10.1134/S0006297923110202

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