Allopregnanolone Promotes Migration and Invasion of Human Glioblastoma Cells through the Protein Tyrosine Kinase c-Src Activation

Abstract

Glioblastomas (GBs) are the most aggressive and common primary malignant brain tumors. Steroid hormone progesterone (P4) and its neuroactive metabolites, such as allopregnanolone (3α-THP) are synthesized by neural, glial, and malignant GB cells. P4 promotes cellular proliferation, migration, and invasion of human GB cells at physiological concentrations. It has been reported that 3α-THP promotes GB cell proliferation. Here we investigated the effects of 3α-THP on GB cell migration and invasion, the participation of the enzymes involved in its metabolism (AKR1C1-4), and the role of the c-Src kinase in 3α-THP effects in GBs. 3α-THP 100 nM promoted migration and invasion of U251, U87, and LN229 human-derived GB cell lines. We observed that U251, LN229, and T98G cell lines exhibited a higher protein content of AKR1C1-4 than normal human astrocytes. AKR1C1-4 silencing did not modify 3α-THP effects on migration and invasion. 3α-THP activated c-Src protein at 10 min (U251 cells) and 15 min (U87 and LN229 cells). Interestingly, the pharmacological inhibition of c-Src decreases the promoting effects of 3α-THP on cell migration and invasion. Together, these data indicate that 3α-THP promotes GB migration and invasion through c-Src activation.

Progesterone and its metabolite allopregnanolone promote invasion of human glioblastoma cells through metalloproteinase‑9 and cSrc kinase

“…The results of the present study demonstrated that inhibition of cSrc activity decreased invasiveness in both U251 and U87 cell lines, as measured using a Matrigel Boyden chamber assay. A similar effect of 3α-THP has also been recently reported by our laboratory ( 18 ).…”

“…It has recently been reported that 3α-THP promotes the migration of glioblastoma cells independently of the oxidation of other P4 metabolites with high binding affinity from the PR. In addition, the activation of cSrc kinase by 3a-THP has been reported ( 18 , 42 ). Melfi et al ( 19 ) reported that, in rat Schwann cells, induced cell migration was dependent on cSrc activation.…”

“…4390844) (both Thermo Fisher Scientific, Inc.) was used to evaluate the effect of PR on MMP-2 and −9 expression 48 h after the addition of siRNAs. The transfection protocol was performed as previously described ( 15 , 18 ).…”

“…In addition, finasteride, an inhibitor of 5α-reductase, the rate-limiting enzyme for the P4 transformation to 3α-THP, partially inhibited this effect ( 17 ). Moreover, 3α-THP regulates migration in glioblastoma cells, and this effect is dependent on the activation of cSrc ( 18 ). In rat Schwann cells, the 3α-THP induction of cell migration is regulated by cSrc and Fak ( 19 ).…”

Neuroactive Steroids, Toll-like Receptors, and Neuroimmune Regulation: Insights into Their Impact on Neuropsychiatric Disorders

“…Moreover, allopregnanolone has been shown to promote cell proliferation in a human glioblastoma (GB) U87 multiforme cell line and the expression of genes associated with tumor progression, including TGF-β1, epidermal growth factor receptor, vascular endothelial growth factor, and cylin-D1 [512]. Additionally, allopregnanolone was shown to increase the migration and invasion of GB cells, potentially through the activation of the cellular proto-oncogene tyrosine-protein kinase Src pathway [513]. These results are further supported by the fact that sex steroid hormones, including estrogens and progestins, can induce the progression of GBs [514,515].…”