Objective      To determine the effects of fluorouracil and mitomycin on the autophagy

aging

and apoptosis in cholangiocarcinoma cell line QBC939

and investigate the relationship of autophagy

aging

and apoptosis

and the underlying possible mechanisms.       Methods      Fluorouracil (25 to 200 μg/mL)

mitomycin (0.25

to 2.0 μg/mL) and their corresponding final concentration of the mixed solution were respectively used to treat QBC939 cells for 0

24 and 48 h

respectively. Cell counting kit-8 (CCK-8) assay was employed to measure cell viability and determine the combined chemotherapy drug concentration by detecting of cell proliferation. Cell apoptosis was detected by flow cytometry with Annexin V-FITC staining

aging by SA-β-Gal staining

and autophagy by flow cytometry with monodansylcadaverin (MDC) staining. Above detections on proliferation

aging

apoptosis and autophagy were carried out again by the same methods after the apoptosis and autophagy were blocked by caspase inhibitor Z-VAD-FMK and 3-methyl adenine (3-MA) respectively.       Results      QBC939 proliferation was significantly inhibited when combination of fluorouracil and mitomycin (with final concentrations of 100 and 1.0 μg/mL respectively) for 24 h

though widespread cell death was not observed. There was significant differences among groups (F=26.929

P

0.05)

and compared with the control group

the autophagy

aging and apoptosis were changed significantly (

P

0.01). After the treatment of autophagy inhibitor

the level of autophagy was reduced

cell proliferation was inhibited

the amount of senescent cells was decreased

and apoptosis was significantly increased. On the other hand

after the treatment of apoptosis inhibitor

cell apoptosis was suppressed

proliferation was enhanced

cell aging was significantly increased

but no significant change was found in autophagy.       Conclusion      Combination treatment of fluorouracil and mitomycin significantly not only reduces cell proliferation but also induces apoptosis in cholangiocarcinoma cells. These changes were related with autophagy in cholangiocarcinoma cells after chemotherapy. Autophagy might be a mechanism for QBC939 cells to avoid adverse environment.