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Anti-cancer activity of Psoralea fructus through the downregulation of cyclin D1 and CDK4 in human colorectal cancer cells

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ABSTRACT

Background: Psoralea Fructus (PF), the dried and ripe fruit of Psoralea corylifolia exhibits an anti-cancer activity. However, the molecular mechanisms by which PF inhibits the proliferation of cancer cells have not been elucidated in detail. Cyclin D1 and CDK4 are important regulatory proteins in cell growth and are overexpressed in many cancer cells. In this study, we investigated the molecular mechanism of PF on the downregulation of cyclin D1 and CDK4 level.

Methods: Cell growth was evaluated by MTT assay. The effect of PF on cyclin D1 and CDK4 expression was evaluated by Western blot or RT-PCR.

Results: PF suppressed the proliferation of human colorectal cancer cell lines such as HCT116 (IC50: 45.3 ± 1.2 μg/ml), SW480 (IC50: 37.9 ± 1.6 μg/ml), LoVo (IC50: 23.3 ± 1.9 μg/ml μg/ml) HT-29 (IC50 value: 40.7 ± 1.5 μg/ml). PF induced decrease in the protein expression of cyclin D1 and CDK4. However, the mRNA expression of cyclin D1 and CDK4 did not be changed by PF; rather it suppressed the expression of cyclin D1 and CDK4 via the proteasomal degradation. In cyclin D1 degradation, we found that T286 of cyclin D1 play a pivotal role in PF-mediated cyclin D1 degradation. Subsequent experiments with several kinase inhibitors suggest that PF-mediated degradation of cyclin D1 and CDK4 is dependent on ERK1/2 and/or GSK3β.

Conclusions: Our results suggest that PF has potential to be a candidate for the development of chemoprevention or therapeutic agents for human colorectal cancer.

No MeSH data available.


Related in: MedlinePlus

The effect on PF on the cell proliferation in human colorectal cancer cells. The cells were plated overnight and then treated with PF for 24 h. Cell proliferation was measured using MTT assay as described in Materials and methods. *P < 0.05 compared to cell without PF treatment
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Fig1: The effect on PF on the cell proliferation in human colorectal cancer cells. The cells were plated overnight and then treated with PF for 24 h. Cell proliferation was measured using MTT assay as described in Materials and methods. *P < 0.05 compared to cell without PF treatment

Mentions: To evaluate whether PF affects the proliferation of human colorectal cancer cells, MTT assay was performed. As shown in Fig. 1, PF treatment for 24 h suppressed the cell growth of HCT116 and SW480 cells by 38 % and 47 % at 25 μg/ml, 54 % and 61 % at 50 μg/ml, and 63 % and 70 % at 100 μg/ml, respectively. In addition, the proliferation of HT-29 and LoVo cells was inhibited by PF treatment at 42 % and 56 % at 25 μg/ml, 69 % and 68 % at 50 μg/ml, and 79 % and 74 % at 100 μg/ml, respectively. Since cell growth inhibition is related to cell cycle arrest, we investigated the expression of cyclin D1 and CDK4 involved in cell cycle progression. The cells were treated with 12.5 and 25 μg/ml for 24 h and Western blot was performed. As shown in Fig. 2a-d, the proteins of cyclin D1 and CDK4 were down-regulated by PF treatment. In time-course experiment (Fig. 2e), the protein expression of cyclin D1 started to be decreased at 1 h after PF treatment, while CDK4 expression was suppressed at 10 h after PF treatment.Fig. 1


Anti-cancer activity of Psoralea fructus through the downregulation of cyclin D1 and CDK4 in human colorectal cancer cells
The effect on PF on the cell proliferation in human colorectal cancer cells. The cells were plated overnight and then treated with PF for 24 h. Cell proliferation was measured using MTT assay as described in Materials and methods. *P < 0.05 compared to cell without PF treatment
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5037586&req=5

Fig1: The effect on PF on the cell proliferation in human colorectal cancer cells. The cells were plated overnight and then treated with PF for 24 h. Cell proliferation was measured using MTT assay as described in Materials and methods. *P < 0.05 compared to cell without PF treatment
Mentions: To evaluate whether PF affects the proliferation of human colorectal cancer cells, MTT assay was performed. As shown in Fig. 1, PF treatment for 24 h suppressed the cell growth of HCT116 and SW480 cells by 38 % and 47 % at 25 μg/ml, 54 % and 61 % at 50 μg/ml, and 63 % and 70 % at 100 μg/ml, respectively. In addition, the proliferation of HT-29 and LoVo cells was inhibited by PF treatment at 42 % and 56 % at 25 μg/ml, 69 % and 68 % at 50 μg/ml, and 79 % and 74 % at 100 μg/ml, respectively. Since cell growth inhibition is related to cell cycle arrest, we investigated the expression of cyclin D1 and CDK4 involved in cell cycle progression. The cells were treated with 12.5 and 25 μg/ml for 24 h and Western blot was performed. As shown in Fig. 2a-d, the proteins of cyclin D1 and CDK4 were down-regulated by PF treatment. In time-course experiment (Fig. 2e), the protein expression of cyclin D1 started to be decreased at 1 h after PF treatment, while CDK4 expression was suppressed at 10 h after PF treatment.Fig. 1

View Article: PubMed Central - PubMed

ABSTRACT

Background: Psoralea Fructus (PF), the dried and ripe fruit of Psoralea corylifolia exhibits an anti-cancer activity. However, the molecular mechanisms by which PF inhibits the proliferation of cancer cells have not been elucidated in detail. Cyclin D1 and CDK4 are important regulatory proteins in cell growth and are overexpressed in many cancer cells. In this study, we investigated the molecular mechanism of PF on the downregulation of cyclin D1 and CDK4 level.

Methods: Cell growth was evaluated by MTT assay. The effect of PF on cyclin D1 and CDK4 expression was evaluated by Western blot or RT-PCR.

Results: PF suppressed the proliferation of human colorectal cancer cell lines such as HCT116 (IC50: 45.3&thinsp;&plusmn;&thinsp;1.2&nbsp;&mu;g/ml), SW480 (IC50: 37.9&thinsp;&plusmn;&thinsp;1.6&nbsp;&mu;g/ml), LoVo (IC50: 23.3&thinsp;&plusmn;&thinsp;1.9&nbsp;&mu;g/ml&nbsp;&mu;g/ml) HT-29 (IC50 value: 40.7&thinsp;&plusmn;&thinsp;1.5&nbsp;&mu;g/ml). PF induced decrease in the protein expression of cyclin D1 and CDK4. However, the mRNA expression of cyclin D1 and CDK4 did not be changed by PF; rather it suppressed the expression of cyclin D1 and CDK4 via the proteasomal degradation. In cyclin D1 degradation, we found that T286 of cyclin D1 play a pivotal role in PF-mediated cyclin D1 degradation. Subsequent experiments with several kinase inhibitors suggest that PF-mediated degradation of cyclin D1 and CDK4 is dependent on ERK1/2 and/or GSK3&beta;.

Conclusions: Our results suggest that PF has potential to be a candidate for the development of chemoprevention or therapeutic agents for human colorectal cancer.

No MeSH data available.


Related in: MedlinePlus