The Study On Human Lung Cancer Cell Line A-549 By The Eicosapentaenoic Acid Plus Carboplatin

Purpose To observe change of cell proliferation and induction of apoptosis on human lung cancer cell line A-549 by the combination of eicosapentaenoic acid (EPA) and carboplatin, in order to prove that Anti-cancer effect of carboplatin was increased by EPA and its mechanism of action. Provide experimental evidence to explore the possibility of the clinical dose reduction of carboplatin to reduce toxicity.Methods MTT assay was used to determine the effect of EPA, carboplatin and their combination on human lung cancer cell line A-549 anti-proliferation.To observe human lung cancer cell line A-549 morphological changes of apoptosis by inverted microscope and HE staining and quantitative analysis of apoptosis by flow cytometry (FCM). Microplate assay was used to determine Reactive oxygen species (ROS) level of human lung cancer cell line A-549. To observe the expression of bcl-2 by immunocytochemistry (ICC).Results①MTT assay results showed that human lung cancer A-549 cell proliferation had been reduced in a dose dependent manner by EPA.②Carboplatin, EPA and combination groups were seen tumor cells were markedly reduced, in part became smaller in size and round, nuclear condensation,lost connections and lung cancer cells original form by inverted microscope and HE staining.It accorded with morphological changes of apoptosis.③Flow cytometry results showed that the apoptosis rate of combination group than carboplatin group significantly increased.④After 48h treatment, ROS lever of carboplatin group and control group was no significant difference, ROS lever of EPA group and combined group were obviously increased.⑤The expression of anti-apoptotic factor bcl-2 of combined group was significantly reduced than the control group, EPA group and carboplatin group by observing the results of immunocytochemical staining. Conclusions①MTT assay results showed that human lung cancer A-549 cell proliferation had been reduced in a dose dependent manner by EPA.②EPA significantly enhanced effect of anti-tumor proliferation of carboplatin.③The apoptosis rate of combination group than carboplatin group significantly increased.④ROS lever of combined group were obviously increased.⑤The expression of anti-apoptotic factor bcl-2 of combined group was significantly reduced than the control group, EPA group and carboplatin group.