Dissertation
Dissertation > Medicine, health > Chinese Medicine > TCM oncology

Experimental Research on Melittin in Inhibiting Human Gastric Cell Line BGC-823and the Synergistic Interaction between Melittin and Chemotherapeutic Agents

Author HuangShuRan
Tutor WangRuiPing
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
CLC R273
Type Master's thesis
Year 2012
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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.

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