Experimental Research on Melittin in Inhibiting Human Gastric Cell Line BGC-823and the Synergistic Interaction between Melittin and Chemotherapeutic Agents
|School||Nanjing University of Traditional Chinese Medicine|
|Course||TCM medical oncology|
|Keywords||melittin human gastric cell line BGC-823 cell proliferation cell adhesion cell invasion early apoptosis cell cycle synergy effect|
Objective:This study was designed to explore:①The inhibiting effect of melittin in vitro on the proliferation of human gastric cell line BGC-823;②The effect of melittin in vitro on adhesion and invasion capability of BGC-823cells;③The effect of melittin on the apoptosis and cell cycle of BGC-823;④To assess the interaction between melittin and chemotherapeutic agents widely used in human gastric cell line BGC-823treatmen.Method:①MTT method was used to observe the effects of different concentrations of melittin on human gastric cell line BGC-823proliferation and the relationship between drμg effect and time;②Adhesion capability of BGC-823cells was evaluated also by MTT method;③Transwell invasion assay was used to observe the invasion capability;④Flow cytometry, AnnexinV/PI double staining method was used to observe the early apoptosis induced by different concentrations of melittin in human gastric cell line BGC-823;⑤After BGC-823cells were treated with melittin at different concentrations cell cycle were determined by flow cytometry (FCM);⑥Synergistic interaction on human gastric cancer BGC-823cells was evaluated using the combination index (CI) method;⑦Expression of chemotherapeutic agent-associated genes of BGC-823cells with or without treatment were measured by realtime quantitative PCR.Resμlts:①Melittin inhibited the proliferation of human gastric cell line BGC-823with a certain degree of concentration-dependence:no significant inhibition was observed when the drμg concentration was2μg/ml; it significantly inhibited the proliferation when the drμg concentration increased to32μg/ml and the inhibition gradually increased with time.②The adhesion and invasion capability of BGC-823cells decreased after treatment of melittin.③Melittin applied on human gastric cell line BGC-823for6h, induced the apoptosis of human gastric cell line BGC-823with a certain degree of concentration-dependence.④As compared with the negative control group, when the cells were treated by16μg/ml of melittin, the percentage of the cells in S phase increased in treatment group, while the percentage of the cells in Go/G1phase decreased, which showed the cells were arrested at the Go/Gi phase of the cell cycle with the increased rate of cell apoptosis.⑤Melittin had a synergistic effect on the cytotoxicity of chemotherapeutic agents in human gastric cell line BGC-823with a certain degree, chemotherapeutic agent-associated genes of BGC-823cells was suppressed expression after treatment.Conclusions:①Melittin inhibited the proliferation of human gastric cell line BGC-823.②Melittin suppresses human gastric cell line BGC-823adhesion and invasion capability.③Melittin can induce human gastric cell line BGC-823apoptosis and co μ Id hinder the cell cycle at S phase.④Melittin had a synergistic effect on the cytotoxicity of chemotherapeutic agents, The possible mechanisms might be the downregμlation of chemotherapeutic agent-associated genes.