Objective    To investigate the effects of Wortmannin combined with mitomycin C (MMC) on the proliferation

apoptosis and expressions of related genes Cyclin D

1

and Bad in liver cancer cell line HepG-2.     Methods   HepG-2 cells (10

4

/ml) were treated with 0

25

50

100

200 or 400 nmol/L Wortmannin

0

1.75

3.75

7.5

15 or 30 nmol/L MMC

or combined concentra

tions (0

26.875

53.75

107.5

215 or 430 nmol/L) respectively. Cell suppression rate were detected by MTT assay. Nucleus apoptosis of HepG-2 cells were examined by Hochest 33258 fluorescent staining. Change of cell cycle and apoptosis in HepG-2 cells were detected by flow cytometry (FCM). Expressions of PI3K/Akt signaling transduction pathway related genes and proteins were tested by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis.     Results    The inhibition ratio of cell proliferation was significant higher in combined groups than in either single groups (P0.05). FCM showed HepG-2 cells were arrested at G

0

/G

1

phrase

and apoptotic rate were increased from (3.23±1.32)% in control to (14.82±1.36)% in combined use of Wortmannin and MMC for 24 h (P0.01). Typical changes in apoptotic morphology of HepG-2 cells were observed by fluorescence microscopy. Activation of p-Akt was inhibited

and mRNA and protein levels of Cyclin D

1

were decreased

but those of Bad were increased.     Conclusion    Combined use of Wortmannin and MMC has a synergic antineoplastic effect on HepG-2 cells.