Evaluation of the p53 pathway in polycystic ovarian syndrome pathogenesis and apoptosis enhancement in human granulosa cells through transcriptome data analysis

Abstract

The polycystic ovarian syndrome (PCOS) is closely associated with enhanced apoptosis of granulosa cells, which have a vital role in maturation of oocytes. p53 plays a critical role in the regulation of apoptosis and cell cycle arrest, metabolism and insulin resistance. The aim of this study was to investigate the impact of p53 pathway in enhancing apoptosis and abnormal function of granulosa cells. In this study, microarray analysis and RNA sequencing were downloaded from the GEO and used as datasets. Principal Component Analysis (PCA) and online SSizer tool were applied to evaluate the experiment quality control and sample sufficiency, respectively. Bioinformatics’ analyses were performed on the selected datasets, and validated by qRT-PCR and western blot analyses. Three datasets out of five ones were chosen for re-analyzing based on the PCA outcomes. 21 deregulated genes were identified via filters including p < 0.05 and |log2FC|≥ 1. Functional enrichment analysis confirmed the relevance of cell cycle regulation and apoptosis as common biological hallmarks in PCOS. Results have shown differentially expressed p53 target genes involved in apoptosis (BAX, FAS, PMAIP1, and CASP8), cell cycle (Cyclins, Cyclin dependent kinases), glucose metabolism and insulin resistance (THBS1), and p53 regulation (MDM2). Subsequently, the relative mRNA expression of FAS, PMAIP1 and MDM2 genes, and protein levels of p53 and MDM2 were confirmed using granulosa cells collected from 20 PCOS women and 18 control individuals by qRT-PCR and western blot, respectively. Results of this study represent the possible role of p53 pathway in pathogenesis of PCOS particularly, through the enhancement of apoptosis in granulosa cells.

Maternal MicroRNA Profile Changes When LH Is Added to the Ovarian Stimulation Protocol: A Pilot Study

“…The p53 gene family member (TP53 in humans) is a guardian of the female germline and plays a key role in determining the oocyte fate upon damage [ 34 ]. P53 is involved in many cellular processes, including steroid hormone regulation, and its role in the pathogenesis of polycystic ovarian syndrome (PCOS)—a complex endocrinologic disorder with increasing risk of infertility, ovulatory disfunction, hyperandrogenism and raised LH:FSH ratio [ 35 ]—has been proposed. Recent investigations of increased rates of embryonic implantation failure and miscarriage in women with polymorphisms in the p53 gene could demonstrate the importance of p53 in female fertility and IVF success [ 36 ].…”

Programmed cell death 4: A novel player in the pathogenesis of polycystic ovary syndrome

“…The main suppressor of p53, that is, MDM2, also showed downregulation, leading to a decrease in the ubiquitin-dependent degradation of p53, and, consequently an increase in its cellular expression and transcriptional activity. 70 The T A B L E 2 Association of PDCD4 expression with cell cycle progression, proliferation, and apoptosis.…”

Dysfunction of Human Estrogen Signaling as a Novel Molecular Signature of Polycystic Ovary Syndrome

“…These preantral follicles showed a higher survival capacity [2,3], with granulosa cells (GCs) responding prematurely to the luteinizing hormone (LH) [4] and exhibiting a higher proliferation rate than normal [5]. However, when examining follicles at a later stage of development, such as preovulatory follicles obtained from women undergoing in vitro fertilization (IVF) protocols, several studies reported a significant increase in the apoptosis capacity of GCs in PCOS [6][7][8][9][10].…”