Alpha-santalol, a chemopreventive agent against skin cancer, causes G2/M cell cycle arrest in both p53-mutated human epidermoid carcinoma A431 cells and p53 wild-type human melanoma UACC-62 cells.
BACKGROUND
alpha-Santalol, an active component of sandalwood oil, has shown chemopreventive effects on skin cancer in different murine models. However, effects of alpha-santalol on cell cycle have not been studied. Thus, the objective of this study was to investigate effects of alpha-santalol on cell cycle progression in both p53 mutated human epidermoid carcinoma A431 cells and p53 wild-type human melanoma UACC-62 cells to elucidate the mechanism(s) of action.


METHODS
MTT assay was used to determine cell viability in A431 cells and UACC-62; fluorescence-activated cell sorting (FACS) analysis of propidium iodide staining was used for determining cell cycle distribution in A431 cells and UACC-62 cells; immunoblotting was used for determining the expression of various proteins and protein complexes involved in the cell cycle progression; siRNA were used to knockdown of p21 or p53 in A431 and UACC-62 cells and immunofluorescence microscopy was used to investigate microtubules in UACC-62 cells.


RESULTS
alpha-Santalol at 50-100 muM decreased cell viability from 24 h treatment and alpha-santalol at 50 muM-75 muM induced G2/M phase cell cycle arrest from 6 h treatment in both A431 and UACC-62 cells. alpha-Santalol altered expressions of cell cycle proteins such as cyclin A, cyclin B1, Cdc2, Cdc25c, p-Cdc25c and Cdk2. All of these proteins are critical for G2/M transition. alpha-Santalol treatment up-regulated the expression of p21 and suppressed expressions of mutated p53 in A431 cells; whereas, alpha-santalol treatment increased expressions of wild-type p53 in UACC-62 cells. Knockdown of p21 in A431 cells, knockdown of p21 and p53 in UACC-62 cells did not affect cell cycle arrest caused by alpha-santalol. Furthermore, alpha-santalol caused depolymerization of microtubules similar to vinblastine in UACC-62 cells.


CONCLUSIONS
This study for the first time identifies effects of alpha-santalol in G2/M phase arrest and describes detailed mechanisms of G2/M phase arrest by this agent, which might be contributing to its overall cancer preventive efficacy in various mouse skin cancer models.
Zhang
Xiaoying
X
Department of Pharmaceutical Sciences, South Dakota State University, Brookings, SD 57007, USA. Chandradhar.Dwivedi@sdstate.edu.
Chen
Wei
W
Guillermo
Ruth
R
Chandrasekher
Gudiseva
G
Kaushik
Radhey S
RS
Young
Alan
A
Fahmy
Hesham
H
Dwivedi
Chandradhar
C