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Yang-Dan-Tang, Identified from 15 Chinese Herbal Formulae, Inhibits Human Lung Cancer Cell Proliferation via Cell Cycle Arrest.

Wang TC, Fang CN, Shen CC, Wei HY, Weng YP, Lin JY, Hsieh-Li HM, Lee CY - Evid Based Complement Alternat Med (2012)

Bottom Line: We identified three formulae (Formulae 3, 5, and 14) that substantially decreased the survival of A549 cells but did not affect MRC5 normal lung tissue cells.Flow cytometry analysis showed that treatment of Formula 14 induced cell cycle arrest in G1 and G2 phase without causing significant cell death.These results were also confirmed by Western blot analysis, with decreased expression of G1/S and G2/M promoting cell cycle machinery including cyclin D3, cyclin B1, CDK4, and CDK6.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Science, National Taiwan Normal University, Taipei 116, Taiwan.

ABSTRACT
Lung cancer has long been one of the most deadly forms of cancer. The majority of lung cancers are of the non-small-cell lung cancer (NSCLC) type. Here we used the non-small-cell lung carcinoma cell line A549 to screen 15 different traditional Chinese herbal medicine (CHM) formulae to explore the possible mechanisms of alternative medicine in lung cancer therapy. We identified three formulae (Formulae 3, 5, and 14) that substantially decreased the survival of A549 cells but did not affect MRC5 normal lung tissue cells. Formula 14, Yang-Dan-Tang, a modified decoction of Ramulus Cinnamomi Cassiae, was chosen for further characterization. Flow cytometry analysis showed that treatment of Formula 14 induced cell cycle arrest in G1 and G2 phase without causing significant cell death. These results were also confirmed by Western blot analysis, with decreased expression of G1/S and G2/M promoting cell cycle machinery including cyclin D3, cyclin B1, CDK4, and CDK6. This study provides further insight into the possible working mechanism of Yang-Dan-Tang in patients.

No MeSH data available.


Related in: MedlinePlus

Western blots of the G1/S and G2/M regulatory proteins in A549 cells after 72 hr treatment of Formula 14. (a) Examination of A549 G1/S regulatory protein levels after treatment with Formula 14 for 72 hr. The expression levels of cyclin D3, CDK4, and CDK6 and the phosphorylation of Rb were significantly decreased after treatment with Formula 14. (b) Western blots of cyclin B1, phosphorylated cdc2, and total cdc2. The expression levels of cyclin B1 and the phosphorylation of cdc2 were decreased, while the expression of cdc2 was increased after treatment with Formula 14. Results of 3 independent experiments were shown at the same time to show the consistent results. Student's t-test: *P < 0.05. (n = 3).
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fig5: Western blots of the G1/S and G2/M regulatory proteins in A549 cells after 72 hr treatment of Formula 14. (a) Examination of A549 G1/S regulatory protein levels after treatment with Formula 14 for 72 hr. The expression levels of cyclin D3, CDK4, and CDK6 and the phosphorylation of Rb were significantly decreased after treatment with Formula 14. (b) Western blots of cyclin B1, phosphorylated cdc2, and total cdc2. The expression levels of cyclin B1 and the phosphorylation of cdc2 were decreased, while the expression of cdc2 was increased after treatment with Formula 14. Results of 3 independent experiments were shown at the same time to show the consistent results. Student's t-test: *P < 0.05. (n = 3).

Mentions: We then examined the expression of cell cycle regulatory proteins to uncover more molecular clues to the effects of Formula 14. Western blot analyses showed that expression of G1/S transition promoting proteins, including cyclin D3, D-type-cyclin-dependent kinase CDK4 and CDK6, and phosphorylated retinoblastoma protein (pRb), the substrate of cyclin D/CDK4/6 complex, was decreased by Formula 14 (Figure 5(a)). The reduced levels of these molecules could result in G1 phase arrest, which is in agreement with the result of the previous cell cycle analysis. In addition to the G1 phase arrest, the G2 phase cell population was also increased in A549 cells after treatment with Formula 14 for 72 hr (Figure 3(a)). We thus further checked the G2/M transition regulating proteins, cyclin B1 and cdc2 (cdk1). Treatment with Formula 14 decreased the expression of both cyclin B1 and phosphorylated cdc2 (Tyr15-p-cdc2) and increased the total amount of all forms of cdc2 (Figure 5(b)). The downregulation of cyclin B1 is in accordance with the G2 arrest effect of Formula 14, while the decreased ratio of Tyr15-p-cdc2/total cdc2 may result from the mixed composition of Formula 14.


Yang-Dan-Tang, Identified from 15 Chinese Herbal Formulae, Inhibits Human Lung Cancer Cell Proliferation via Cell Cycle Arrest.

Wang TC, Fang CN, Shen CC, Wei HY, Weng YP, Lin JY, Hsieh-Li HM, Lee CY - Evid Based Complement Alternat Med (2012)

Western blots of the G1/S and G2/M regulatory proteins in A549 cells after 72 hr treatment of Formula 14. (a) Examination of A549 G1/S regulatory protein levels after treatment with Formula 14 for 72 hr. The expression levels of cyclin D3, CDK4, and CDK6 and the phosphorylation of Rb were significantly decreased after treatment with Formula 14. (b) Western blots of cyclin B1, phosphorylated cdc2, and total cdc2. The expression levels of cyclin B1 and the phosphorylation of cdc2 were decreased, while the expression of cdc2 was increased after treatment with Formula 14. Results of 3 independent experiments were shown at the same time to show the consistent results. Student's t-test: *P < 0.05. (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3369556&req=5

fig5: Western blots of the G1/S and G2/M regulatory proteins in A549 cells after 72 hr treatment of Formula 14. (a) Examination of A549 G1/S regulatory protein levels after treatment with Formula 14 for 72 hr. The expression levels of cyclin D3, CDK4, and CDK6 and the phosphorylation of Rb were significantly decreased after treatment with Formula 14. (b) Western blots of cyclin B1, phosphorylated cdc2, and total cdc2. The expression levels of cyclin B1 and the phosphorylation of cdc2 were decreased, while the expression of cdc2 was increased after treatment with Formula 14. Results of 3 independent experiments were shown at the same time to show the consistent results. Student's t-test: *P < 0.05. (n = 3).
Mentions: We then examined the expression of cell cycle regulatory proteins to uncover more molecular clues to the effects of Formula 14. Western blot analyses showed that expression of G1/S transition promoting proteins, including cyclin D3, D-type-cyclin-dependent kinase CDK4 and CDK6, and phosphorylated retinoblastoma protein (pRb), the substrate of cyclin D/CDK4/6 complex, was decreased by Formula 14 (Figure 5(a)). The reduced levels of these molecules could result in G1 phase arrest, which is in agreement with the result of the previous cell cycle analysis. In addition to the G1 phase arrest, the G2 phase cell population was also increased in A549 cells after treatment with Formula 14 for 72 hr (Figure 3(a)). We thus further checked the G2/M transition regulating proteins, cyclin B1 and cdc2 (cdk1). Treatment with Formula 14 decreased the expression of both cyclin B1 and phosphorylated cdc2 (Tyr15-p-cdc2) and increased the total amount of all forms of cdc2 (Figure 5(b)). The downregulation of cyclin B1 is in accordance with the G2 arrest effect of Formula 14, while the decreased ratio of Tyr15-p-cdc2/total cdc2 may result from the mixed composition of Formula 14.

Bottom Line: We identified three formulae (Formulae 3, 5, and 14) that substantially decreased the survival of A549 cells but did not affect MRC5 normal lung tissue cells.Flow cytometry analysis showed that treatment of Formula 14 induced cell cycle arrest in G1 and G2 phase without causing significant cell death.These results were also confirmed by Western blot analysis, with decreased expression of G1/S and G2/M promoting cell cycle machinery including cyclin D3, cyclin B1, CDK4, and CDK6.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Science, National Taiwan Normal University, Taipei 116, Taiwan.

ABSTRACT
Lung cancer has long been one of the most deadly forms of cancer. The majority of lung cancers are of the non-small-cell lung cancer (NSCLC) type. Here we used the non-small-cell lung carcinoma cell line A549 to screen 15 different traditional Chinese herbal medicine (CHM) formulae to explore the possible mechanisms of alternative medicine in lung cancer therapy. We identified three formulae (Formulae 3, 5, and 14) that substantially decreased the survival of A549 cells but did not affect MRC5 normal lung tissue cells. Formula 14, Yang-Dan-Tang, a modified decoction of Ramulus Cinnamomi Cassiae, was chosen for further characterization. Flow cytometry analysis showed that treatment of Formula 14 induced cell cycle arrest in G1 and G2 phase without causing significant cell death. These results were also confirmed by Western blot analysis, with decreased expression of G1/S and G2/M promoting cell cycle machinery including cyclin D3, cyclin B1, CDK4, and CDK6. This study provides further insight into the possible working mechanism of Yang-Dan-Tang in patients.

No MeSH data available.


Related in: MedlinePlus