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. 2012:2012:326096.
doi: 10.1155/2012/326096. Epub 2012 Jun 7.

6-Gingerol Inhibits Growth of Colon Cancer Cell LoVo via Induction of G2/M Arrest

Affiliations

6-Gingerol Inhibits Growth of Colon Cancer Cell LoVo via Induction of G2/M Arrest

Ching-Bin Lin et al. Evid Based Complement Alternat Med. 2012.

Abstract

6-Gingerol, a natural component of ginger, has been widely reported to possess antiinflammatory and antitumorigenic activities. Despite its potential efficacy against cancer, the anti-tumor mechanisms of 6-gingerol are complicated and remain sketchy. In the present study, we aimed to investigate the anti-tumor effects of 6-gingerol on colon cancer cells. Our results revealed that 6-gingerol treatment significantly reduced the cell viability of human colon cancer cell, LoVo, in a dose-dependent manner. Further flow cytometric analysis showed that 6-gingerol induced significant G2/M phase arrest and had slight influence on sub-G1 phase in LoVo cells. Therefore, levels of cyclins, cyclin-dependent kinases (CDKs), and their regulatory proteins involved in S-G2/M transition were investigated. Our findings revealed that levels of cyclin A, cyclin B1, and CDK1 were diminished; in contrast, levels of the negative cell cycle regulators p27(Kip1) and p21(Cip1) were increased in response to 6-gingerol treatment. In addition, 6-gingerol treatment elevated intracellular reactive oxygen species (ROS) and phosphorylation level of p53. These findings indicate that exposure of 6-gingerol may induce intracellular ROS and upregulate p53, p27(Kip1), and p21(Cip1) levels leading to consequent decrease of CDK1, cyclin A, and cyclin B1 as result of cell cycle arrest in LoVo cells. It would be suggested that 6-gingerol should be beneficial to treatment of colon cancer.

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Figures

Figure 1
Figure 1
6-Gingerol inhibited the cell viability of LoVo cells. Cells were treated with indicated concentration of 6-gingerol for 24 h or 48 h, and the cell viability was analyzed by MTT assay. Data were shown as the means ± SD. Three independent experiments were performed for statistical analysis. *P < 0.05 and **P < 0.005 as compared to control (C).
Figure 2
Figure 2
6-Gingerol induced G2/M phase arrest of LoVo cells. Cells were treated with indicated concentration of 6-gingerol for 24 h or 48 h, and the percentages of various cell cycle phases, including sub-G1, G0/G1, S, and G2/M, were analyzed and quantitated by flow cytometry. Data were shown as the means ± SD. Three independent experiments were performed for statistical analysis. *P < 0.05 and **P < 0.005 as compared to corresponding control.
Figure 3
Figure 3
Effects of 6-gingerol on activation of caspase 3 and caspase 8 of LoVo cells. Cells were treated with indicated concentration of 6-gingerol for 24 h or 48 h, and then the cell lysates were subjected to immunoblot for detection of caspase 3 and caspase 8. Protein levels were relatively quantitated by densitometric analysis using GAPDH as control.
Figure 4
Figure 4
Effects of 6-gingerol on CDK1 and cyclins of LoVo cells. Cells were treated with indicated concentration of 6-gingerol for 24 h or 48 h, and then the cell lysates were subjected to immunoblot for detection of indicated proteins. Protein levels were relatively quantitated by densitometric analysis using GAPDH as control.
Figure 5
Figure 5
Effects of 6-gingerol on p21Cip1 and p27Kip1 of LoVo cells. Cells were treated with indicated concentration of 6-gingerol for 24 h or 48 h, and then the cell lysates were subjected to immunoblot for detection of p21Cip1 and p27Kip1. Protein levels were relatively quantitated by densitometric analysis using GAPDH as control.
Figure 6
Figure 6
Effects of 6-gingerol on p53 and intracellular ROS of LoVo cells. (a) Cells were treated with indicated concentration of 6-gingerol for 24 h or 48 h, and then the cell lysates were subjected to immunoblot for detection of p53. Protein levels were relatively quantitated by densitometric analysis using GAPDH as control. (b) Cells were treated with indicated concentration of 6-gingerol for 24 h, and the intracellular ROS was determined as described in the Section 2.

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