Effects of rosmarinic acid on proliferation and apoptosis of HCT-8 cells and its molecular mechanisms / 中草药

ABSTRACT

Objective: To investigate the effects of rosmarinic acid (RosA) on the proliferation and apoptosis of human colon cancer HCT-8 cells, and explore the related mechanisms. Methods: The proliferation of HCT-8 cells was detected by CCK-8 assay at different concentrations of RosA for different time periods. The apoptosis rates of HCT-8 cells and the expression of related proteins were investigated after the treatment of RosA at 15, 45, and 75 μmol/L for 72 h according to the CCK-8 results. The apoptosis rates of HCT-8 cells were detected by FCM. The mRNA expressions of p53, Bax, and Puma were detected by RT-qPCR. The protein levels of p53, Bax, Puma, and active Caspase-9 were detected by Western blotting. Results: RosA could inhibit the proliferation of human colon cancer cells HCT-8 in a time- and dose-dependent manner. RosA at 15 and 45 μmol/L mainly induced early apoptosis (P < 0.01), RosA at 75 μmol/L mainly induced late apoptosis (P < 0.01). RosA could up-regulate the mRNA expression of Puma in a dose-dependent manner. RosA at 45 and 75 μmol/L increased the mRNA expression of p53 and Bax (P < 0.05). RosA could up-regulate the protein levels of Puma, Bax, and active Caspase-9 in a dose-dependent manner. RosA at 75 μmol/L could significantly increase the protein expression of p53 (P < 0.05). Conclusion: RosA can significantly inhibit the proliferation of HCT-8 cells by inducing the apoptosis. The apoptosis-inducing proteins of p53, Puma, Bax, and active Caspase-9 induce the apoptosis of cells.