Published online Aug 21, 2013. doi: 10.3748/wjg.v19.i31.5159
Revised: July 4, 2013
Accepted: July 12, 2013
Published online: August 21, 2013
Processing time: 90 Days and 19 Hours
AIM: To investigate the effects of suberoylanilide hydroxamic acid (SAHA) on proliferation and apoptosis of a human hepatocellular carcinoma cell line (HepG2.2.15) and hepatitis B virus (HBV) replication.
METHODS: HepG2.2.15 cells were treated with different concentrations of SAHA. Cell morphology was examined by confocal laser scanning microscopy, and cell proliferation was determined using a MTT colorimetric assay. Flow cytometry was used to detect apoptosis and determine cell cycle phase, while hepatitis B surface antigen and hepatitis B e antigen content were measured using chemiluminescence. Reverse transcription polymerase chain reaction was performed to measure HBV DNA in cell lysate.
RESULTS: Cell proliferation rates were significantly reduced by the addition of SAHA. The inhibitory effect of SAHA on cell proliferation was both time- and dose-dependent. After 24 h of treatment with SAHA, the early cell apoptotic rate increased from 3.25% to 21.02% (P = 0.041). The proportion of G0/G1 phase cells increased from 50.3% to 65.3% (P = 0.039), while that of S phase cells decreased from 34.9% to 20.6% (P = 0.049). After 48 h of treatment, hepatitis B surface antigen and hepatitis B e antigen content increased from 12.33 ± 0.62 to 25.42 ± 2.67 (P = 0.020) and 28.92 ± 1.24 to 50.48 ± 1.85 (P = 0.026), respectively. Furthermore, HBV DNA content increased from 4.54 ± 0.46 to 8.34 ± 0.59 (P = 0.029).
CONCLUSION: SAHA inhibits HepG2.2.15 cell proliferation, promotes apoptosis, and stimulates HBV replication. In combination with anti-HBV drugs, SAHA may potentially be used cautiously for treatment of hepatocellular carcinoma.
Core tip: HepG2.2.15 cells were treated with different concentrations of suberoylanilide hydroxamic acid (SAHA). Hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) content were measured using chemiluminescence. Reverse transcription polymerase chain reaction was performed to measure hepatitis B virus (HBV) DNA in cell lysate. Results found that, the inhibitory effect of SAHA on cell proliferation was both time- and dose-dependent. After 24 h of treatment, the early cell apoptotic rate increased. After 48 h of treatment, HBsAg and HBeAg content both increased. Furthermore, HBV DNA content increased. In combination with anti-HBV drugs, SAHA may potentially be used cautiously for treatment of hepatocellular carcinoma.