Activation of membrane progesterone receptor-alpha increases proliferation, migration, and invasion of human glioblastoma cells

Glioblastomas are the most common and aggressive primary brain tumors in adults, and patients with glioblastoma have a median survival of 15 months. Some alternative therapies, such as Src family kinase inhibitors, have failed presumably because other signaling pathways compensate for their effects. In the last ten years, it has been proven that sex hormones such as progesterone (P4) can induce growth, migration, and invasion of glioblastoma cells through its intracellular progesterone receptor (PR), which is mostly known for its role as a transcription factor, but it can also induce non-genomic actions. These non-classic actions are, in part, a consequence of its interaction with cSrc, which plays a significant role in the progression of glioblastomas. We studied the relation between PR and cSrc, and its effects in human glioblastoma cells. Our results showed that P4 and R5020 (specific PR agonist) activated cSrc protein since both progestins increased the p-cSrc (Y416)/cSrc ratio in U251 and U87 human glioblastoma derived cell lines. When siRNA against the PR gene was used, the activation of cSrc by P4 was abolished. The co-immunoprecipitation assay showed that cSrc and PR interact in U251 cells. P4 treatment also promoted the increase in the p-Fak (Y397) (Y576/577)/Fak and the decrease in p-Paxillin (Y118)/Paxillin ratio, which are significant components of the focal adhesion complex and essential for migration and invasion processes. A siRNA against cSrc gene blocked the increase in the p-Fak (Y576/Y577)/Fak ratio and the migration induced by P4, but not the decrease in p-Paxillin (Y118)/Paxillin ratio. We analyzed the potential role of cSrc over PR phosphorylation in three databases, and one putative tyrosine residue in the amino acid 87 of PR was found. Our results showed that P4 induces the activation of cSrc protein through its PR. The latter and cSrc could interact in a bidirectional mode for regulating the activity of proteins involved in migration and invasion of glioblastomas.

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Jinlong capsule inhibits migration and invasion in human glioblastoma cells via the modulation of mTOR/S6 signaling pathway

Drug Design Development and Therapy ◽

10.2147/dddt.s195409 ◽

2019 ◽

Vol Volume 13 ◽

pp. 1023-1032 ◽

Cited By ~ 1

Author(s):

Jingren Shi ◽

Wenli Zhang ◽

Lu He ◽

Fanhong Kong ◽

Meichen Pan ◽

...

Keyword(s):

Signaling Pathway ◽

Migration And Invasion ◽

Glioblastoma Cells ◽

Human Glioblastoma ◽

Human Glioblastoma Cells

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Shikonin Inhibits the Migration and Invasion of Human Glioblastoma Cells by Targeting Phosphorylated β-Catenin and Phosphorylated PI3K/Akt: A Potential Mechanism for the Anti-Glioma Efficacy of a Traditional Chinese Herbal Medicine

International Journal of Molecular Sciences ◽

10.3390/ijms161023823 ◽

2015 ◽

Vol 16 (10) ◽

pp. 23823-23848 ◽

Cited By ~ 36

Author(s):

Feng-Ying Zhang ◽

Yi Hu ◽

Zhong-You Que ◽

Ping Wang ◽

Yun-Hui Liu ◽

...

Keyword(s):

Herbal Medicine ◽

Chinese Herbal Medicine ◽

Migration And Invasion ◽

Potential Mechanism ◽

Glioblastoma Cells ◽

Human Glioblastoma ◽

Traditional Chinese Herbal Medicine ◽

Chinese Herbal ◽

Human Glioblastoma Cells

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Approaches to the design of selective ligands for membrane progesterone receptor alpha

Biochemistry (Moscow) ◽

10.1134/s0006297913030048 ◽

2013 ◽

Vol 78 (3) ◽

pp. 236-243 ◽

Cited By ~ 6

Author(s):

O. V. Lisanova ◽

T. A. Shchelkunova ◽

I. A. Morozov ◽

P. M. Rubtsov ◽

I. S. Levina ◽

...

Keyword(s):

Progesterone Receptor ◽

Receptor Alpha ◽

Selective Ligands ◽

Membrane Progesterone Receptor ◽

Membrane Progesterone Receptor Alpha

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Progesterone-induced activation of membrane-bound progesterone receptors in murine macrophage cells

Journal of Endocrinology ◽

10.1530/joe-14-0470 ◽

2014 ◽

Vol 224 (2) ◽

pp. 183-194 ◽

Cited By ~ 23

Author(s):

Jing Lu ◽

Joshua Reese ◽

Ying Zhou ◽

Emmet Hirsch

Keyword(s):

Progesterone Receptor ◽

Inflammatory Responses ◽

Progesterone Receptors ◽

Murine Macrophage ◽

Induced Expression ◽

Receptor Alpha ◽

Downstream Target ◽

Macrophage Cells ◽

Membrane Progesterone Receptor ◽

Membrane Progesterone Receptor Alpha

Parturition is an inflammatory process mediated to a significant extent by macrophages. Progesterone (P4) maintains uterine quiescence in pregnancy, and a proposed functional withdrawal of P4 classically regulated by nuclear progesterone receptors (nPRs) leads to labor. P4 can affect the functions of macrophages despite the reported lack of expression of nPRs in these immune cells. Therefore, in this study we investigated the effects of the activation of the putative membrane-associated PR on the function of macrophages (a key cell for parturition) and discuss the implications of these findings for pregnancy and parturition. In murine macrophage cells (RAW 264.7), activation of mPRs by P4 modified to be active only extracellularly by conjugation to BSA (P4BSA, 1.0×10−7 mol/l) caused a pro-inflammatory shift in the mRNA expression profile, with significant upregulation of the expression of cyclooxygenase 2 (COX2 (Ptgs2)), Il1B, and Tnf and downregulation of membrane progesterone receptor alpha (Paqr7) and oxytocin receptor (Oxtr). Pretreatment with PD98059, a MEK1/2 inhibitor, significantly reduced P4BSA-induced expression of mRNA of Il1B, Tnf, and Ptgs2. Inhibition of protein kinase A (PKA) by H89 blocked P4BSA-induced expression of Il1B and Tnf mRNA. P4BSA induced rapid phosphorylation of MEK1/2 and CREB (a downstream target of PKA). This phosphorylation was inhibited by pretreatment with PD98059 and H89, respectively, revealing that MEK1/2 and PKA are two of the components involved in mPR signaling. Taken together, these results indicate that changes in membrane progesterone receptor alpha expression and signaling in macrophages are associated with the inflammatory responses; and that these changes might contribute to the functional withdrawal of P4 related to labor.

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Proliferative and Invasive Effects of Progesterone-Induced Blocking Factor in Human Glioblastoma Cells

BioMed Research International ◽

10.1155/2017/1295087 ◽

2017 ◽

Vol 2017 ◽

pp. 1-10 ◽

Cited By ~ 11

Author(s):

Araceli Gutiérrez-Rodríguez ◽

Valeria Hansberg-Pastor ◽

Ignacio Camacho-Arroyo

Keyword(s):

Migration And Invasion ◽

Glioblastoma Cells ◽

Human Glioblastoma ◽

Astrocytoma Cells ◽

Blocking Factor ◽

Human Glioblastoma Cells ◽

U87 Cells ◽

Antagonist Ru486 ◽

Human Astrocytoma Cells ◽

Invasion Assays

Progesterone-induced blocking factor (PIBF) is a progesterone (P4) regulated protein expressed in different types of high proliferative cells including astrocytomas, the most frequent and aggressive brain tumors. It has been shown that PIBF increases the number of human astrocytoma cells. In this work, we evaluated PIBF regulation by P4 and the effects of PIBF on proliferation, migration, and invasion of U87 and U251 cells, both derived from human glioblastomas. PIBF mRNA expression was upregulated by P4 (10 nM) from 12 to 24 h. Glioblastoma cells expressed two PIBF isoforms, 90 and 57 kDa. The content of the shorter isoform was increased by P4 at 24 h, while progesterone receptor antagonist RU486 (10 μM) blocked this effect. PIBF (100 ng/mL) increased the number of U87 cells on days 4 and 5 of treatment and induced cell proliferation on day 4. Wound-healing assays showed that PIBF increased the migration of U87 (12–48 h) and U251 (24 and 48 h) cells. Transwell invasion assays showed that PIBF augmented the number of invasive cells in both cell lines at 24 h. These data suggest that PIBF promotes proliferation, migration, and invasion of human glioblastoma cells.

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