Cannabinoid exposure does not alter estradiol biosynthesis in human KGN granulosa cells

Steroidogenesis is essential for ovarian physiology and reproductive health. Regulated by hormonal signals, it is susceptible to external modulators, notably environmental exposures. As cannabis becomes more accessible globally, its use among women of reproductive age has increased, yet the implications for reproductive endocrinology remain poorly understood and contradictory. In this study, we investigated whether cannabinoids modulate basal or stimulated estradiol secretion in the human granulosa cell line KGN. To characterize the endocannabinoid system (ECS) in these cells, we performed a meta-analysis of publicly available RNA sequencing datasets, revealing expression of key ECS components. KGN cells were cultured with or without cannabinoids in the presence of protein kinase activators, PKA (FSK), PKB (SC79), and PKC (PMA). Following cannabinoid and kinase stimulations, the media were collected and analyzed for estradiol concentrations via ELISA. We observed no significant changes in basal or activated estradiol secretion in response to THC or CBD. These findings were supported by RT-qPCR analysis showing no alteration in the expression of CYP19A1, the gene encoding aromatase, which catalyzes the conversion of androgens to estrogens in granulosa cells. Although cannabinoids have been shown to influence sex hormones in vivo, our data suggest that these effects are not mediated at the granulosa cell level. This study contributes to a better understanding of how cannabinoids may interact with ovarian steroidogenesis and reproductive function.