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Caffeic acid phenethyl ester induced cell cycle arrest and growth inhibition in androgen-independent prostate cancer cells via regulation of Skp2, p53, p21Cip1 and p27Kip1.

Lin HP, Lin CY, Huo C, Hsiao PH, Su LC, Jiang SS, Chan TM, Chang CH, Chen LT, Kung HJ, Wang HD, Chuu CP - Oncotarget (2015)

Bottom Line: CAPE treatment decreased Skp2 and Akt1 protein expression in LNCaP 104-R1 tumors as compared to control group.Overexpression of Skp2, or siRNA knockdown of p21Cip1, p27Kip1, or p53 blocked suppressive effect of CAPE treatment.Co-treatment of CAPE with PI3K inhibitor LY294002 or Bcl-2 inhibitor ABT737 showed synergistic suppressive effects.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan, ROC.

ABSTRACT
Prostate cancer (PCa) patients receiving the androgen ablation therapy ultimately develop recurrent castration-resistant prostate cancer (CRPC) within 1-3 years. Treatment with caffeic acid phenethyl ester (CAPE) suppressed cell survival and proliferation via induction of G1 or G2/M cell cycle arrest in LNCaP 104-R1, DU-145, 22Rv1, and C4-2 CRPC cells. CAPE treatment also inhibited soft agar colony formation and retarded nude mice xenograft growth of LNCaP 104-R1 cells. We identified that CAPE treatment significantly reduced protein abundance of Skp2, Cdk2, Cdk4, Cdk7, Rb, phospho-Rb S807/811, cyclin A, cyclin D1, cyclin H, E2F1, c-Myc, SGK, phospho-p70S6kinase T421/S424, phospho-mTOR Ser2481, phospho-GSK3α Ser21, but induced p21Cip1, p27Kip1, ATF4, cyclin E, p53, TRIB3, phospho-p53 (Ser6, Ser33, Ser46, Ser392), phospho-p38 MAPK Thr180/Tyr182, Chk1, Chk2, phospho-ATM S1981, phospho-ATR S428, and phospho-p90RSK Ser380. CAPE treatment decreased Skp2 and Akt1 protein expression in LNCaP 104-R1 tumors as compared to control group. Overexpression of Skp2, or siRNA knockdown of p21Cip1, p27Kip1, or p53 blocked suppressive effect of CAPE treatment. Co-treatment of CAPE with PI3K inhibitor LY294002 or Bcl-2 inhibitor ABT737 showed synergistic suppressive effects. Our finding suggested that CAPE treatment induced cell cycle arrest and growth inhibition in CRPC cells via regulation of Skp2, p53, p21Cip1, and p27Kip1.

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Micro-Western Array image and heatmap of abundance and phosphorylation fold changes of signaling proteins in LNCaP 104-R1 cells treated with CAPE(A) LNCaP 104-R1 cells were treated with 0, 10, 20, 40 μM CAPE for 96 h. Micro-Western Arrays were performed to measure the changes in abundance and modification of total Akt, Akt1, Akt2, phospho-Akt Thr308, phospho-Akt Ser473, AR, Cdk2, cyclin A, cyclin D1, c-Myc, p21Cip1, p27Kip1, Skp2, p70S6 kinase, phospho-p70 S6 kinase Thr421/Ser424, p90RSK, phospho-p90RSK Ser380, Rb, phospho-Rb Ser807/811, PTEN, Bcl-2, fatty acid synthase (FAS), p53, phospho-p53 Ser392, phospho-p53 Ser6, phospho-p53 Ser33, phospho-p53 Ser46, GSK3α, GSK3β, phospho-GSK3α Ser21, phospho-Gsk3β Ser9, SGK, phospho-SGK Ser255/Thr241, CKI, CKIIα, mTOR, phospho-mTOR Ser2448, PDK1, phospho-PDK1 Ser241, phospho-mTOR ser2481, p38 MAPK, phospho-p38 MAPK Thr180/Tyr182, NF-κB p65, and NF-κB p50. Protein abundance of α-tubulin and β-actin was used as loading control. Red color and green color indicated 680 nM and 780 nM wavelength detected by Licor Odyssey scanner for rabbit antibodies and mouse antibodies, respectively. Blue arrows indicated the correct location of band for the detected proteins. (B) Proteins were organized in the y-axis of the heatmap based on time of maximal fold change amplitude. Green color indicated decrease of protein expression while red color indicated increase of protein expression under treatment of CAPE.
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Figure 5: Micro-Western Array image and heatmap of abundance and phosphorylation fold changes of signaling proteins in LNCaP 104-R1 cells treated with CAPE(A) LNCaP 104-R1 cells were treated with 0, 10, 20, 40 μM CAPE for 96 h. Micro-Western Arrays were performed to measure the changes in abundance and modification of total Akt, Akt1, Akt2, phospho-Akt Thr308, phospho-Akt Ser473, AR, Cdk2, cyclin A, cyclin D1, c-Myc, p21Cip1, p27Kip1, Skp2, p70S6 kinase, phospho-p70 S6 kinase Thr421/Ser424, p90RSK, phospho-p90RSK Ser380, Rb, phospho-Rb Ser807/811, PTEN, Bcl-2, fatty acid synthase (FAS), p53, phospho-p53 Ser392, phospho-p53 Ser6, phospho-p53 Ser33, phospho-p53 Ser46, GSK3α, GSK3β, phospho-GSK3α Ser21, phospho-Gsk3β Ser9, SGK, phospho-SGK Ser255/Thr241, CKI, CKIIα, mTOR, phospho-mTOR Ser2448, PDK1, phospho-PDK1 Ser241, phospho-mTOR ser2481, p38 MAPK, phospho-p38 MAPK Thr180/Tyr182, NF-κB p65, and NF-κB p50. Protein abundance of α-tubulin and β-actin was used as loading control. Red color and green color indicated 680 nM and 780 nM wavelength detected by Licor Odyssey scanner for rabbit antibodies and mouse antibodies, respectively. Blue arrows indicated the correct location of band for the detected proteins. (B) Proteins were organized in the y-axis of the heatmap based on time of maximal fold change amplitude. Green color indicated decrease of protein expression while red color indicated increase of protein expression under treatment of CAPE.

Mentions: As CAPE treatment reduced cell proliferation and induced cell cycle arrest in CRPC cells, we used Micro-Western Arrays (MWAs), a high-throughput Western blotting assay [19, 22, 26], to determine how proteins regulating cell proliferation, cell survival, and cell cycle progression are affected by CAPE treatment. CAPE treatment significantly decreased protein levels of fatty acid synthase (FAS), retinoblastoma protein (Rb), phospho-Rb Ser807/811, c-Myc, p70S6kinase, phospho-p70S6kinase Thr421/Ser424, Skp2, p90RSK, and NF-κB p65. Alternatively, CAPE treatment significantly increased p53, phospho-p53 Ser392, phospho-p53 Ser33, phospho-p53 S6, phospho-p53 Ser46, p27Kip1, mTOR, CK1, GSK3α, CK2α, cyclin A, p38 MAPK, and p21Cip1 (Figure 5A, 5B).


Caffeic acid phenethyl ester induced cell cycle arrest and growth inhibition in androgen-independent prostate cancer cells via regulation of Skp2, p53, p21Cip1 and p27Kip1.

Lin HP, Lin CY, Huo C, Hsiao PH, Su LC, Jiang SS, Chan TM, Chang CH, Chen LT, Kung HJ, Wang HD, Chuu CP - Oncotarget (2015)

Micro-Western Array image and heatmap of abundance and phosphorylation fold changes of signaling proteins in LNCaP 104-R1 cells treated with CAPE(A) LNCaP 104-R1 cells were treated with 0, 10, 20, 40 μM CAPE for 96 h. Micro-Western Arrays were performed to measure the changes in abundance and modification of total Akt, Akt1, Akt2, phospho-Akt Thr308, phospho-Akt Ser473, AR, Cdk2, cyclin A, cyclin D1, c-Myc, p21Cip1, p27Kip1, Skp2, p70S6 kinase, phospho-p70 S6 kinase Thr421/Ser424, p90RSK, phospho-p90RSK Ser380, Rb, phospho-Rb Ser807/811, PTEN, Bcl-2, fatty acid synthase (FAS), p53, phospho-p53 Ser392, phospho-p53 Ser6, phospho-p53 Ser33, phospho-p53 Ser46, GSK3α, GSK3β, phospho-GSK3α Ser21, phospho-Gsk3β Ser9, SGK, phospho-SGK Ser255/Thr241, CKI, CKIIα, mTOR, phospho-mTOR Ser2448, PDK1, phospho-PDK1 Ser241, phospho-mTOR ser2481, p38 MAPK, phospho-p38 MAPK Thr180/Tyr182, NF-κB p65, and NF-κB p50. Protein abundance of α-tubulin and β-actin was used as loading control. Red color and green color indicated 680 nM and 780 nM wavelength detected by Licor Odyssey scanner for rabbit antibodies and mouse antibodies, respectively. Blue arrows indicated the correct location of band for the detected proteins. (B) Proteins were organized in the y-axis of the heatmap based on time of maximal fold change amplitude. Green color indicated decrease of protein expression while red color indicated increase of protein expression under treatment of CAPE.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4466643&req=5

Figure 5: Micro-Western Array image and heatmap of abundance and phosphorylation fold changes of signaling proteins in LNCaP 104-R1 cells treated with CAPE(A) LNCaP 104-R1 cells were treated with 0, 10, 20, 40 μM CAPE for 96 h. Micro-Western Arrays were performed to measure the changes in abundance and modification of total Akt, Akt1, Akt2, phospho-Akt Thr308, phospho-Akt Ser473, AR, Cdk2, cyclin A, cyclin D1, c-Myc, p21Cip1, p27Kip1, Skp2, p70S6 kinase, phospho-p70 S6 kinase Thr421/Ser424, p90RSK, phospho-p90RSK Ser380, Rb, phospho-Rb Ser807/811, PTEN, Bcl-2, fatty acid synthase (FAS), p53, phospho-p53 Ser392, phospho-p53 Ser6, phospho-p53 Ser33, phospho-p53 Ser46, GSK3α, GSK3β, phospho-GSK3α Ser21, phospho-Gsk3β Ser9, SGK, phospho-SGK Ser255/Thr241, CKI, CKIIα, mTOR, phospho-mTOR Ser2448, PDK1, phospho-PDK1 Ser241, phospho-mTOR ser2481, p38 MAPK, phospho-p38 MAPK Thr180/Tyr182, NF-κB p65, and NF-κB p50. Protein abundance of α-tubulin and β-actin was used as loading control. Red color and green color indicated 680 nM and 780 nM wavelength detected by Licor Odyssey scanner for rabbit antibodies and mouse antibodies, respectively. Blue arrows indicated the correct location of band for the detected proteins. (B) Proteins were organized in the y-axis of the heatmap based on time of maximal fold change amplitude. Green color indicated decrease of protein expression while red color indicated increase of protein expression under treatment of CAPE.
Mentions: As CAPE treatment reduced cell proliferation and induced cell cycle arrest in CRPC cells, we used Micro-Western Arrays (MWAs), a high-throughput Western blotting assay [19, 22, 26], to determine how proteins regulating cell proliferation, cell survival, and cell cycle progression are affected by CAPE treatment. CAPE treatment significantly decreased protein levels of fatty acid synthase (FAS), retinoblastoma protein (Rb), phospho-Rb Ser807/811, c-Myc, p70S6kinase, phospho-p70S6kinase Thr421/Ser424, Skp2, p90RSK, and NF-κB p65. Alternatively, CAPE treatment significantly increased p53, phospho-p53 Ser392, phospho-p53 Ser33, phospho-p53 S6, phospho-p53 Ser46, p27Kip1, mTOR, CK1, GSK3α, CK2α, cyclin A, p38 MAPK, and p21Cip1 (Figure 5A, 5B).

Bottom Line: CAPE treatment decreased Skp2 and Akt1 protein expression in LNCaP 104-R1 tumors as compared to control group.Overexpression of Skp2, or siRNA knockdown of p21Cip1, p27Kip1, or p53 blocked suppressive effect of CAPE treatment.Co-treatment of CAPE with PI3K inhibitor LY294002 or Bcl-2 inhibitor ABT737 showed synergistic suppressive effects.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan, ROC.

ABSTRACT
Prostate cancer (PCa) patients receiving the androgen ablation therapy ultimately develop recurrent castration-resistant prostate cancer (CRPC) within 1-3 years. Treatment with caffeic acid phenethyl ester (CAPE) suppressed cell survival and proliferation via induction of G1 or G2/M cell cycle arrest in LNCaP 104-R1, DU-145, 22Rv1, and C4-2 CRPC cells. CAPE treatment also inhibited soft agar colony formation and retarded nude mice xenograft growth of LNCaP 104-R1 cells. We identified that CAPE treatment significantly reduced protein abundance of Skp2, Cdk2, Cdk4, Cdk7, Rb, phospho-Rb S807/811, cyclin A, cyclin D1, cyclin H, E2F1, c-Myc, SGK, phospho-p70S6kinase T421/S424, phospho-mTOR Ser2481, phospho-GSK3α Ser21, but induced p21Cip1, p27Kip1, ATF4, cyclin E, p53, TRIB3, phospho-p53 (Ser6, Ser33, Ser46, Ser392), phospho-p38 MAPK Thr180/Tyr182, Chk1, Chk2, phospho-ATM S1981, phospho-ATR S428, and phospho-p90RSK Ser380. CAPE treatment decreased Skp2 and Akt1 protein expression in LNCaP 104-R1 tumors as compared to control group. Overexpression of Skp2, or siRNA knockdown of p21Cip1, p27Kip1, or p53 blocked suppressive effect of CAPE treatment. Co-treatment of CAPE with PI3K inhibitor LY294002 or Bcl-2 inhibitor ABT737 showed synergistic suppressive effects. Our finding suggested that CAPE treatment induced cell cycle arrest and growth inhibition in CRPC cells via regulation of Skp2, p53, p21Cip1, and p27Kip1.

Show MeSH
Related in: MedlinePlus