Transcriptomic analysis of gene cascades involved in protein kinase A and C signaling in the KGN line of human ovarian granulosa tumor cells†

Abstract

The developmental competence of an oocyte is its capacity to resume maturation, undergo successful fertilization, and reach the blastocyst stage. This competence is acquired through interaction with somatic cells of the follicle. Cumulus and granulosa cells support oocyte development, while the oocyte influences follicular cell growth and differentiation. Studies suggest that follicle-stimulating hormone and luteinizing hormone play an essential role in oocyte competence acquisition through signaling initiated by protein kinases A and C (PKA and PKC) in granulosa cells. Using a microarray and RT-qPCR, the transcriptome of human granulosa-like tumor cells (KGN) treated for 24 h with forskolin (FSK) or phorbol 12-myristate 13-acetate (PMA) was analyzed to determine the effects of PKA and PKC stimulation on gene expression. Protein-kinase-driven signaling appeared to involve five major upstream regulators, namely epidermal growth factor (EGF), transforming growth factor beta 1 (TGFβ1), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGF2), and hepatocyte growth factor (HGF). Gene associations with seven major ovarian functions were identified: Prostaglandin- endoperoxide synthase 2 (PTGS2), interleukin 8 (IL8), and interleukin 6 (IL6) with inflammation; Steroidogenic acute regulatory protein (STAR), cytochrome P450scc (CYP11A1), and cytochrome P450 family 19 subfamily A member 1 (CYP19A1) with steroidogenesis; Vascular endothelial growth factor C (VEGFC), Vascular endothelial growth factor A (VEGFA), and C-X-C chemokine receptor type 4 (CXCR4) with angiogenesis; Amphiregulin (AREG), epidermal growth factor receptor (EGFR), and sprouty RTK signaling antagonist 2 (SPRY2) with differentiation, BCL2 associated X (BAX), BCL2 like 12 (BCL2L12), and caspase 1(CASP1) with apoptosis, Cyclin D1 (CCND1), cyclin B1 (CCNB1), and cyclin B2 (CCNB2) with division; and Matrix metalloproteinase-1 (MMP1), Matrix metallopeptidase 9 (MMP9), and TIMP metallopeptidase inhibitor 1 (TIMP1) with ovulation. Overall, these results indicate that signaling via both PKA and PKC potentiates gene regulation of functions such as inflammation and apoptosis, while functions such as differentiation, ovulation and angiogenesis are partial to one kinase or the other. These results improve understanding of the pathways underlying the most important changes that occur in the follicle prior to ovulation.

Human amnion-derived mesenchymal stem cell (hAD-MSC) transplantation improves ovarian function in rats with premature ovarian insufficiency (POI) at least partly through a paracrine mechanism

“…Although follicular growth is chiefly regulated by gonadotropins in a variety of monovular and polyovular species [45, 46], the regulation of certain local growth factors, such as VEGF, FGF2, HGF, and IGF-1, is inevitable, without which follicles fail to grow further [47–49]. Gonadotropins also stimulate ovarian follicular angiogenesis partly by regulating the expression of those growth factors [47, 50], and angiogenesis is an important prerequisite for follicular development and maturation [51]. Additionally, those growth factors have been shown to stimulate cell proliferation and have antiapoptotic effects in GCs [48, 52], and GCs support oocyte survival and are required for follicular development [51].…”

“…Gonadotropins also stimulate ovarian follicular angiogenesis partly by regulating the expression of those growth factors [47, 50], and angiogenesis is an important prerequisite for follicular development and maturation [51]. Additionally, those growth factors have been shown to stimulate cell proliferation and have antiapoptotic effects in GCs [48, 52], and GCs support oocyte survival and are required for follicular development [51]. VEGF is a key angiogenic factor that has been demonstrated to play an important role in the growth, recruitment, and establishment of dominance of antral follicles in buffalo, cattle, and humans [53, 54].…”

“…Studies have shown that FGF2 may promote GC survival and steroidogenesis in an autocrine and paracrine manner [59]. FGF2 may be responsible for GC proliferation and the prevention of apoptosis and may play an important role in follicular development by affecting cell differentiation and ovulation processes [51]. IGF-1, a potent stimulator of cell proliferation and differentiation, regulates GC apoptosis and steroidogenesis during follicular development [60].…”

Comparative analysis of ovarian transcriptomes between prolific and non‐prolific goat breeds via high‐throughput sequencing

“…The MSMB expression was reduced in ovarian cancer and correlates to survival (Henriksen, Lundwall, Udby, & Fernlund, ), indicating that MSMB appears to be necessary for normal function of ovary. TIMP1 was identified as one of the important genes associated with ovulation (Tremblay & Sirard, ). In the mouse, SCARB1 was expressed in cytoplasm and periphery of theca, granulosa and cumulus cells of developing follicles.…”

FSH receptor binding inhibitor impacts K-Ras and c-Myc of ovarian cancer and signal pathway

“…Follicle stimulating hormone (FSH) is an important ovarian epithelial cell growth-promoting factor [ 6 , 7 ]. FSH functions via binding to its cognate receptor (FSHR).…”