Figure 4.
Anticancer drugs induced autophagy in lung cancer cells. A, A549 cells (2 × 105) were treated with the chemotherapeutic drugs cisplatin (2 μg/mL), LBH589 (100 nmol/L), or rapamycin (100 nmol/L), or a combination of two drugs for 24 h. The mRNA levels of ATG5, ATG7 and BECN1 were assayed by RT‐qPCR. B, chemotherapeutic drug‐induced cells were pre‐treated with or without Bafilomycin A1 (BafA1, 20 nmol/L) for 1 h followed by Western blot using anti‐LC3 antibody. C, A549 cells were treated with chemotherapeutic drugs for 48 h after transfection with pEGFP‐LC3 plasmid. LC3 punctate‐positive cells were observed by fluorescence microscopy and quantitation of the percentage of cells with punctate GFP‐LC3 fluorescence per total GFP‐LC3‐positive cells. Data represent mean ± SD calculated from three experiments of 100 transfected cells each. D, chemotherapeutic drug‐treated cells were stained with AO (1 μg/mL) for 20 min. Autophagic cells were analysed using flow cytometry. Results are representative of at least three independent experiments. *P < .05 and **P < .01 compared with untreated cells