[Effects of hedyotis diffusa on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45].

Objective: To investigate the effects of hedyotis diffusa (injection) on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45 cells. Methods: The human gastric cancer MNK-45 cells were divided into 4 groups, each group was set with 3 replicates. The control group was MNK-45 cells without added hedyotis diffusa; the 3 groups of experimental groups were treated with hedyotis diffusa at final concentrations of 20 , 30, 40 μg / ml respectively; each group was incubated for 48 h in a 5% carbon dioxide incubator, and the morphological changes of the cells were observed under a laser confocal microscope. Mitochondrial membrane potential was detected by flow cytometry. The expressions of Cytochrome C (Cyt c), caspase3 and caspase9 genes and proteins were detected by qRT-PCR and Western blot respectively. Results: Compared with the control group, the mitochondrial membrane potentials of MNK-45 cells were significantly reduced in the hedyotis diffusa treated groups at final concentrations of 20, 30, and 40 μg / ml (P＜0. 01). The gene expressions of Cyt c, caspase3, and caspase9 were significantly up-regulated (P＜0. 01) and their protein expressions were also significantly increased (P＜0. 05 or P＜0. 01). The 40 μg / ml hedyotis diffusa treatment group performed best. Conclusion: In the final concentration range of 20 ~ 40 μg / ml, hedyotis diffusa can reduce human gastric cancer MNK-45 cells mitochondrial membrane potential, induce apoptosis and up-regulate Cyt c, caspase3 and caspase9 gene expressions.