Jichu yixue yu linchuang (Jul 2020)
Catecholamine stress hormones promote the cell cycle re-entry of G0 phase in mouse hepatocarcinoma cell line H22
Abstract
Objective To investigate the effect of stress hormones on cell cycle re-entry of G0 phase in mouse H22 hepatocarcinoma cells. Methods Flow cytometry antibody staining with CD44 and CD133 was used to determine the presence of cancer stem cells in H22 cells. In vitro, after treatment of H22 cells with stress hormone corticosterone (CORT), epinephrine (EPI) and norepinephrine (NE) for 48 h, the proportion changes of cells in G0 phase and cell cycle distribution were detected by Ki-67 and PI flow cytometry staining. The hormone receptor antagonists were used to verify the receptor subtypes that mediated the cell cycle re-entry of G0 phase cells. Results The proportion of cancer stem cells (CD44+CD133+) in H22 cells was 0.80%±0.15%. Compared with control group, the portion of G0 phase cells was unchanged after corticosterone treatment, but was decreased by 62.50% and 53.00% respectively after EPI and NE treatment. Meanwhile, the total cells, proportion of G2/M and S phase cells were increased compared to control group. The protein expression of Skp2 was increased and p27kip1 decreased significantly after EPI and NE treatment(P＜0.05). In addition, α1-adrenergic receptor antagonists terazosin and prazosin blocked the G0 phase cells cycle re-entry effect of EPI and NE. The protein expression of Skp2 was decreased and p27kip1 was increased significantly after prazosin treatment compared to EPI or NE group(P＜0.05). Conclusions Catecholamine stress hormones mediate the cell cycle re-entry of G0 phase cells in mouse H22 hepatocarcinoma cells by activating α1-adrenergic receptor,the mechanism of which may be associated with the Skp2 over-expression and the degradation of p27kip1 protein.
