Objective To investigate the effects of pachymaran on the proliferation, migration and pro-apoptosis of human cervical carcinoma HeLa cells and their related mechanisms.
Methods MTT method was used to detect cell proliferation rate, and we selected the suitable low, medium and high concentrations of pachymaran for experiment. After treatment with low, medium and high concentrations of pachymaran, the morphological changes were observed by inverted phase contrast microscope; nuclear changes were observed by Hoechst 33342 staining; flat plate clone formation test was used to detect the ability of cell clone; cell scratch test was used to detect cell migration ability; flow cytometry was used to detect the apoptosis rate and cell cycle distribution. The expression of related proteins of apoptosis, migration and ERK pathway were detected by Western blot.
Results We selected 30, 40 and 50 mg/ml pachymaran were selected as low, medium and high concentrations in the experiments. After treated with medium and high concentrations of pachymaran, cells and cell nuclei had significant apoptosis morphological changes, but not at low concentration. Different concentrations of pachymaran could reduce the abilities of cell cloning and migration (P < 0.05), increase apoptotic rate (P < 0.05), decreased S phase cells and blocked in G2/M phase (P < 0.05); Cleaved Caspase 3, Cleaved Caspase 8, Cleaved Caspase 9, Bax expression were increased significantly compared with the control group; Bcl-2, MMP-9, VEGFA and p-ERK1/2 expression were decreased significantly (P < 0.05). ERK1/2 expression did not change significantly.
Conclusion Pachymaran could significantly inhibit the proliferation and induce the apoptosis of HeLa cells. The pro-apoptotic mechanism may be related to the inhibition of phosphorylation of ERK signaling pathway by downregulating the expression of p-ERK1/2. In addition, pathymaran could reduce the migration of HeLa cells to a certain extent.
DownLoad: