Dehydroepiandrosterone can promote the apoptosis of KGN in human ovarian granulosa cells

Abstract

Objective To observe the effect of dehydroepiandrosterone (DHEA) on the apoptosis of KGN cell lines. Methods KGN cell line were treated with DHEA at different concentrations (0.01 mmol/L, 0.1 mmol/L, 1 mmol/L, 10 mmol/L). The activity and apoptosis of KGN cells were detected by CCK-8 assay and flow cytometry respectively. Real-time fluorescent quantitative reverse transcriptase polymerase chain reaction (QRT-PCR) was used to detect the expression of androgen receptor (AR), B-cell lymphoma-2 (BCL-2) and BCL-2 associated X protein (BAX) gene mRNA expression levels. The expression of AR, BCL-2, BAX, extracellular regulated protein kinases (ERKs) and phosphorylated ERKs (P-ERKs) were detected by Western blotting. Results Compared with negative control group, cell viability of KGN treated with DHEA decreased significantly (P=0.000), and the apoptosis rate of KGN cells increased with the increase of DHEA dose. The apoptosis of KGN treated with 10 μmol/L DHEA was significantly different from that in negative control group (P=0.019). The expression of BCL-2 protein gradually decreased with the increasing concentration of DHEA. Differences between DHEA 10 mmol/L group and the negative control group were statistically significant (P=0.007). Compared with control group, the expression of BCL-2 mRNA in the cells treated with 0.1 mmol/L and 10 mmol/L DHEA was significantly decreased (P=0.039, P=0.012), while the expression of AR and BAX in the cells did not change significantly (P>0.05). In addition, although the expression level of ERK protein in KGN cells was not significantly different from that in the negative group (P>0.05), the expression of P-ERK protein decreased gradually with the increase of DHEA concentration. The expression of P-ERK protein in 10 mmol/L DHEA-treated group was significantly decreased (P=0.005). Conclusion DHEA can inhibit the growth of KGN cells and promote its apoptosis with the decrease of BCL-2 and P-ERK protein expression, suggesting that DHEA may induce apoptosis of KGN cells through ERK-BCL-2 apoptotic signaling pathway. Key words: Dehydroepiandrosterone (DHEA); KGN cell line; Cell apoptosis; B-cell lymphoma-2 (BCL-2); Phosphorylated ERK (P-ERK)