Synergistic Effect Of Eicosapentaenoic Acid And Vinorelbine On A-549Human Lung Cancer Cell Line

OBJECTIVE To investigate the effect of Eicosapentaenoic Acid (EPA) and its synergistic effect with Vinorelbine (NVB) on cell proliferation and apoptosis and cell cycle of on A-549human lung cancer cell line.METHODS A-549human lung cancer cell line was cultured in vitro,and its morphological changes of apoptosis were observed by using inverted light microscope,HE and Annexin Ⅴ-PI double staining with fluorescence microscope.MTT assay was used to determine the effect of EPA and its synergistic effect with NVB on A-549human lung cancer cells growth inhibitory. The apoptosis and cell cycle of A-549human lung cancer cells exposed to EPA and its synergistic effect with NVB was analyzed by flow cytometry.RESULTS①The A-549human lung cancer cells treated with EPA showed cellular volume small and round,nuclear chromatin condensation, plasma membrance bleb formation, also the apoptotic body formations, loose between the cells and adherent ability decrease under invert microscope.②The A-549human lung cancer cells in treated with EPA group by HE staining showed morphological changes such as cellular volume decrease and round, nuclear chromatin condensation,plasma membrance bleb formation, also the apoptotic body formations were observed under light microscope.③Human A-549lung cancer cells stained by AnnexinV-PI double fluorescent staining were observed under fluorescent microscope,appearing as chromation red condensation in nuclear or green condensation in cells,light green fluorescent apoptotic bodies around cells after induction of EPA.④The MTT test showed that A-549cells were exposed to EPA for24h,48hand72h respectively at final concentrations of15μg/mL～90μg/mL,the growth of cells was inhibited and showed in dose and time dependent manner. EPA combined with NVB showsd significant synergistic anti-tumor effect on A-549cells.⑤Flow cytometry results showed that the apoptosis rate of combination group than NVB group significantly increased.⑥A-549lung cancer cells treated with EPA at final concentration of15μg/mL,30μg/mL,45μg/mL for48h were inhibited and arrested in S phase, the cell percentage of G0/G1phase decrease and that of S phase increase was observed by FCM.A-549lung cancer cells treated with NVB at concentration of15μg/mL for48h were inhibited and arrested in G2/M phase, the cell percentage of G0/G1phase decrease and that of G2/M phase increase was observed by FCM.A-549lung cancer cells treated with Eicosapentaenoic Acid and Vinorelbine at concentration of15μg/mL for48h were inhibited and arrested in S phase and G2/M phase, the cell percentage of G0/G1phase decrease and that of S phase and G2/M phase increase was observed by FCM.CONCLUSIONS1. EPA lead apoptosis of A-549human lung cancer cell line in vitro through morphological assay.2. EPA inhibited growth of A-549human lung cancer cell line in vitro in dose and time dependent manner.3. EPAd and NVB have synergetic effect in the inhibition of A-549human lung cancer cells. Compared with Vinorelbine monotherapy, the combination of NVB and EPA can reduce the dosage of NVB.4. apoptosis rate of combination group than NVB group significantly increased through FCM assay.5. EPA induced S phase arrest and NVB signifi-canfly induced G0/G1phase arrest in A-549cells,which may be one of the mechanisms of EPA and NVB have synergetic efect in the inhibition of A-549human lung cancer cells.