Limits...
Cardamonin Inhibits Metastasis of Lewis Lung Carcinoma Cells by Decreasing mTOR Activity.

Niu PG, Zhang YX, Shi DH, Liu Y, Chen YY, Deng J - PLoS ONE (2015)

Bottom Line: The previous study had proved that the anti-tumor effect of cardamonin was associated with mTOR inhibition.The expression of Snail was decreased by cardamonin, while that of E-cadherin was increased.The metastasis inhibitory effect of cardamonin was correlated with down-regulation of Snail and up-regulation of E-cadherin.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Fujian Provincial Maternal and Child Health Hospital, Fuzhou, Fujian, China.

ABSTRACT
The mammalian target of rapamycin (mTOR) regulates the motility and invasion of cancer cells. Cardamonin is a chalcone that exhibits anti-tumor activity. The previous study had proved that the anti-tumor effect of cardamonin was associated with mTOR inhibition. In the present study, the anti-metastatic effect of cardamonin and its underlying molecule mechanisms were investigated on the highly metastatic Lewis lung carcinoma (LLC) cells. The proliferation, invasion and migration of LLC cells were measured by MTT, transwell and wound healing assays, respectively. The expression and activation of mTOR- and adhesion-related proteins were assessed by Western blotting. The in vivo effect of cardamonin on the metastasis of the LLC cells was investigated by a mouse model. Treated with cardamonin, the proliferation, invasion and migration of LLC cells were significantly inhibited. The expression of Snail was decreased by cardamonin, while that of E-cadherin was increased. In addition, cardamonin inhibited the activation of mTOR and its downstream target ribosomal S6 kinase 1 (S6K1). Furthermore, the tumor growth and its lung metastasis were inhibited by cardamonin in C57BL/6 mice. It indicated that cardamonin inhibited the invasion and metastasis of LLC cells through inhibiting mTOR. The metastasis inhibitory effect of cardamonin was correlated with down-regulation of Snail and up-regulation of E-cadherin.

No MeSH data available.


Related in: MedlinePlus

Cardamonin inhibited the invasion and migration of LLC cells.After incubation with different drugs for 24 h, the cells in the top on the chamber were removed. (A) The cells that had migrated onto the bottom of the chamber were stained with crystal violet and photographed at 200 × magnification. (B) The invaded cells were counted, and the invasive ratio for each group was presented by comparing to the control group. The percentage of the penetrated cells for each group was showed as the bar graph. In the migration assay, a wound was inflicted in the cell layer, and then treated with different drugs for 24 h. (C) The denuded zone was photographed at 100 × magnification. (D) The traveled distance of the different treatment group was expressed as a ratio to the original distance. The migration ratio of each group was showed as the bar graph. The data were presented as the mean ± SD (n = 5). **p < 0.01, compared to the control group.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4440626&req=5

pone.0127778.g003: Cardamonin inhibited the invasion and migration of LLC cells.After incubation with different drugs for 24 h, the cells in the top on the chamber were removed. (A) The cells that had migrated onto the bottom of the chamber were stained with crystal violet and photographed at 200 × magnification. (B) The invaded cells were counted, and the invasive ratio for each group was presented by comparing to the control group. The percentage of the penetrated cells for each group was showed as the bar graph. In the migration assay, a wound was inflicted in the cell layer, and then treated with different drugs for 24 h. (C) The denuded zone was photographed at 100 × magnification. (D) The traveled distance of the different treatment group was expressed as a ratio to the original distance. The migration ratio of each group was showed as the bar graph. The data were presented as the mean ± SD (n = 5). **p < 0.01, compared to the control group.

Mentions: To determine whether cardamonin could function as a new therapeutic compound, we investigate the effect of cardamonin on the cell proliferation. The LLC cells were exposed to different concentrations of cardamonin for 48 h in MTT assay. Consequently, the proliferation of LLC was inhibited by cardamonin in a concentration-dependent manner (Fig 2). However, the inhibitory efficacy of each dose of cardamonin was weaker than that of rapamycin. Invasion is an important step of metastasis. The inhibitory effect of cardamonin on the ability of LLC cells to invade a reconstituted extracellular matrix was assessed by transwell chamber. Our result showed that the number of cells that invaded to the lower chamber was significantly decreased (Fig 3A). Compared with control, the levels of invasion were reduced to 14.8%, 40.7%, 47.2% and 36.8% upon cardamonin and rapamycin treatment, respectively (Fig 3B). To further test the influence of cardamonin on the migration on LLC cells, the scratch assay was implemented. As shown in Fig 3C, a gradual increase of cells in the denuded zone was observed after 24 h in the control and DMSO group. When LLC cells were incubated with cardamonin and rapamycin, the cellular motility were decreased gradually (Fig 3D). The inhibitory effect of invasion and migration was stronger in cardamonin (10 μM) than that in rapamycin (0.1 μM).


Cardamonin Inhibits Metastasis of Lewis Lung Carcinoma Cells by Decreasing mTOR Activity.

Niu PG, Zhang YX, Shi DH, Liu Y, Chen YY, Deng J - PLoS ONE (2015)

Cardamonin inhibited the invasion and migration of LLC cells.After incubation with different drugs for 24 h, the cells in the top on the chamber were removed. (A) The cells that had migrated onto the bottom of the chamber were stained with crystal violet and photographed at 200 × magnification. (B) The invaded cells were counted, and the invasive ratio for each group was presented by comparing to the control group. The percentage of the penetrated cells for each group was showed as the bar graph. In the migration assay, a wound was inflicted in the cell layer, and then treated with different drugs for 24 h. (C) The denuded zone was photographed at 100 × magnification. (D) The traveled distance of the different treatment group was expressed as a ratio to the original distance. The migration ratio of each group was showed as the bar graph. The data were presented as the mean ± SD (n = 5). **p < 0.01, compared to the control group.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440626&req=5

pone.0127778.g003: Cardamonin inhibited the invasion and migration of LLC cells.After incubation with different drugs for 24 h, the cells in the top on the chamber were removed. (A) The cells that had migrated onto the bottom of the chamber were stained with crystal violet and photographed at 200 × magnification. (B) The invaded cells were counted, and the invasive ratio for each group was presented by comparing to the control group. The percentage of the penetrated cells for each group was showed as the bar graph. In the migration assay, a wound was inflicted in the cell layer, and then treated with different drugs for 24 h. (C) The denuded zone was photographed at 100 × magnification. (D) The traveled distance of the different treatment group was expressed as a ratio to the original distance. The migration ratio of each group was showed as the bar graph. The data were presented as the mean ± SD (n = 5). **p < 0.01, compared to the control group.
Mentions: To determine whether cardamonin could function as a new therapeutic compound, we investigate the effect of cardamonin on the cell proliferation. The LLC cells were exposed to different concentrations of cardamonin for 48 h in MTT assay. Consequently, the proliferation of LLC was inhibited by cardamonin in a concentration-dependent manner (Fig 2). However, the inhibitory efficacy of each dose of cardamonin was weaker than that of rapamycin. Invasion is an important step of metastasis. The inhibitory effect of cardamonin on the ability of LLC cells to invade a reconstituted extracellular matrix was assessed by transwell chamber. Our result showed that the number of cells that invaded to the lower chamber was significantly decreased (Fig 3A). Compared with control, the levels of invasion were reduced to 14.8%, 40.7%, 47.2% and 36.8% upon cardamonin and rapamycin treatment, respectively (Fig 3B). To further test the influence of cardamonin on the migration on LLC cells, the scratch assay was implemented. As shown in Fig 3C, a gradual increase of cells in the denuded zone was observed after 24 h in the control and DMSO group. When LLC cells were incubated with cardamonin and rapamycin, the cellular motility were decreased gradually (Fig 3D). The inhibitory effect of invasion and migration was stronger in cardamonin (10 μM) than that in rapamycin (0.1 μM).

Bottom Line: The previous study had proved that the anti-tumor effect of cardamonin was associated with mTOR inhibition.The expression of Snail was decreased by cardamonin, while that of E-cadherin was increased.The metastasis inhibitory effect of cardamonin was correlated with down-regulation of Snail and up-regulation of E-cadherin.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Fujian Provincial Maternal and Child Health Hospital, Fuzhou, Fujian, China.

ABSTRACT
The mammalian target of rapamycin (mTOR) regulates the motility and invasion of cancer cells. Cardamonin is a chalcone that exhibits anti-tumor activity. The previous study had proved that the anti-tumor effect of cardamonin was associated with mTOR inhibition. In the present study, the anti-metastatic effect of cardamonin and its underlying molecule mechanisms were investigated on the highly metastatic Lewis lung carcinoma (LLC) cells. The proliferation, invasion and migration of LLC cells were measured by MTT, transwell and wound healing assays, respectively. The expression and activation of mTOR- and adhesion-related proteins were assessed by Western blotting. The in vivo effect of cardamonin on the metastasis of the LLC cells was investigated by a mouse model. Treated with cardamonin, the proliferation, invasion and migration of LLC cells were significantly inhibited. The expression of Snail was decreased by cardamonin, while that of E-cadherin was increased. In addition, cardamonin inhibited the activation of mTOR and its downstream target ribosomal S6 kinase 1 (S6K1). Furthermore, the tumor growth and its lung metastasis were inhibited by cardamonin in C57BL/6 mice. It indicated that cardamonin inhibited the invasion and metastasis of LLC cells through inhibiting mTOR. The metastasis inhibitory effect of cardamonin was correlated with down-regulation of Snail and up-regulation of E-cadherin.

No MeSH data available.


Related in: MedlinePlus