Study on Forsythin promoting apoptosis of laryngeal carcinoma cells by regulating miRNA-1469

Abstract

Objective

The paper aimed to explore the mechanism of Forsythin regulating miRNA expression in laryngeal carcinoma cells, and to clarify the molecular biological mechanism of Forsythin regulating miRNA to promote the apoptosis of laryngeal carcinoma cells, providing theoretical and experimental basis for clinical application of Forsythin as an anti-laryngeal cancer treatment drug. Methods: A miR-1469 low-expression laryngeal carcinoma cell line was established. Western blot and flow cytometry were applied to detect the effect of Forsythin on cell apoptosis. Western blot was employed to detect the effects of Forsythin on P53 protein, P53 low expression, and P53 overexpression in laryngeal carcinoma cells, as well as the effects on overexpression of miRNA-1469, and on double low expression of P53 and Mcl1. Real-time PCR method was used to detect the effect of miR-1469 on p53 low expression in laryngeal carcinoma cells. Results: Flow cytometry detection of cell apoptosis showed that, after the cells with low miR-1469 expression were treated with Forsythin, the apoptosis rate was significantly reduced. Western blot detection showed that, compared with the Control group, the expression level of miR-1469 was significantly reduced after Forsythin administration in Hep2 cells with low expression of P53. Compared with the idle Control group, the apoptosis level of laryngeal carcinoma cells in Hep2 cells with low expression of P53 was significantly reduced. In Hep2 cells transfected with P53 overexpression plasmids, apoptosis level of laryngeal carcinoma cells increased. Compared with the idle Control group, the apoptosis level of laryngeal carcinoma cells in the single-transformed P53 shRNA group decreased, while the apoptosis level of the double-transformed miR-1469 mimic+P53 shRNA group increased again. After drug treatment, the apoptosis level of the single-transformed P53 shRNA group decreased, while the apoptosis level of the double-transformed Mcl1 shRNA+P53 shRNA group increased again. Conclusion: Forsythin can promote the apoptosis of laryngeal carcinoma cells by up-regulating the expression of miR-1469 and then down-regulating the expression of Mcl1. The drug can up-regulate the expression of miR-1469 by elevating the expression of P53. miR-1469 can promote the apoptosis of laryngeal carcinoma cells by inhibiting the expression of its downstream target gene Mcl1.

Keywords:
forsythin; miRNA-1469; laryngeal carcinoma cells; apoptosis