Korean J Physiol Pharmacol.  2019 Jan;23(1):29-35. 10.4196/kjpp.2019.23.1.29.

Decursin induces apoptosis in glioblastoma cells, but not in glial cells via a mitochondria-related caspase pathway

• Oh ST
• Lee S
• Hua C
• Koo BS
• Pak SC
• Kim DI
• Jeon S
• Shin BA

• KMID: 2443613
• DOI: http://doi.org/10.4196/kjpp.2019.23.1.29

Abstract

Decursin is a major biological active component of Angelica gigas Nakai and is known to induce apoptosis of metastatic prostatic cancer cells. Recently, other reports have been commissioned to examine the anticancer activities of this plant. In this study, we evaluated the inhibitory activity and related mechanism of action of decursin against glioblastoma cell line. Decursin demonstrated cytotoxic effects on U87 and C6 glioma cells in a dose-dependent manner but not in primary glial cells. Additionally, decursin increased apoptotic bodies and phosphorylated JNK and p38 in U87 cells. Decursin also down-regulated Bcl-2 as well as cell cycle dependent proteins, CDK-4 and cyclin D1. Furthermore, decursin-induced apoptosis was dependent on the caspase activation in U87 cells. Taken together, our data provide the evidence that decursin induces apoptosis in glioblastoma cells, making it a potential candidate as a chemotherapeutic drug against brain tumor.

Keyword

Anti-cancer activity; Apoptosis; Cell cycle arrest; Decursin; Glioblastoma

MeSH Terms

Angelica
Apoptosis*
Brain Neoplasms
Cell Cycle
Cell Cycle Checkpoints
Cell Line
Cyclin D1
Extracellular Vesicles
Glioblastoma*
Glioma
Neuroglia*
Plants
Prostatic Neoplasms
Cyclin D1

Figure

• 

  Fig. 1 Decursin has a cytotoxic effect in U87 cells but not in primary glia cells. (A) Chemical structure of decursin. (B) U87 cells were treated with various concentrations of decursin (10, 20, 50, 100 and 200 µM) for 24 h. (C) Primary glia cells were treated with different concentrations of decursin (10, 20, 50, 100 and 200 µM) for 24 h. Cell viability was determined by the MTT assay. Values are mean±SD (n=3). **p<0.01 vs. control, and ***p<0.001 vs. control.

• 

  Fig. 2 Decursin induces apoptosis in U87 cells. (A) The image shows that decursin induced chromatin condensation and apoptotic bodies. (B) U87 cells were treated with 50 µM of decursin for 24 h, stained with Annexin V and PI, and analyzed with FACS system.

• 

  Fig. 3 Decursin induces phosphorylation of JNK and p38 in U87 cells, alters Bcl-2 family protein levels, and activates caspases and PARP-1 in U87 cells. (A) U87 cells were treated with 50 µM of decursin for various periods of time (0, 0.5, 1, 3 and 4 h). The cell lysates were subjected to Western blot analysis for phospho-JNK, total-JNK, phospho-p38 and total p38. (B) U87 cells were treated with 50 or 100 µM of decursin for 24 h. The cell lysates were subjected to Western blot analysis for Bax and Bcl-2 (C), cleaved caspase-3, 7, 9, PARP-1 and β-actin. Values are mean±SD (n=3). *p<0.05 vs. control. **p<0.01 vs. control.

• 

  Fig. 4 Decursin induces apoptosis in a caspase-dependent manner in U87 cells. (A) U87 cells were pre-treated with 5 µM of SB203580 (p38 inhibitor) and SP600125 (JNK inhibitor) for 30 min prior to decursin treatment (50 µM). Cell viability was determined by the MTT assay. (B) U87 cells were pre-treated with 10 µM of pan caspase inhibitor (Cas I) for 30 min prior to decursin treatment (50 µM). Cell viability was determined by the MTT assay. Values are mean±SD (n=4). **p<0.01 vs. control.

• 

  Fig. 5 Decursin increases G1 population of cell cycle and down-regulates cell cycle proteins in U87 cells. U87 cells were treated with 50 µM of decursin for 24 h. The cell lysates were subjected to flow cytometric analysis and Western blot analysis to examine the expression of CDK-4, cyclin D1 and β-actin. Percentage of each cell population was presented in graph (A). The intensity of the each band was normalized to β-actin and presented in bar graphs (B). Values are mean±SD (n=3). **p<0.01 vs. control.

• 

  Fig. 6 Decursin induces apoptosis and caspase-3 activation in C6 cells. (A) C6 cells were treated with various concentrations of decursin (10, 20, 50, 100 and 200 µM) for 24 h and 48 h. Values are mean±SD (n=4). **p<0.01 vs. control, ***p<0.001 vs. control. (B) The cell lysates were subjected to Western blot analysis for cleaved caspase-3 and β-actin.

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