Intest Res.  2017 Apr;15(2):174-181. 10.5217/ir.2017.15.2.174.

Parthenolide promotes apoptotic cell death and inhibits the migration and invasion of SW620 cells

• Liu YC
• Kim SL
• Park YR
• Lee ST
• Kim SW

• KMID: 2425158
• DOI: http://doi.org/10.5217/ir.2017.15.2.174

Abstract

BACKGROUND/AIMS
Parthenolide (PT), a principle component derived from feverfew (Tanacetum parthenium), is a promising anticancer agent and has been shown to promote apoptotic cell death in various cancer cells. In this study, we focused on its functional role in apoptosis, migration, and invasion of human colorectal cancer (CRC) cells.
METHODS
SW620 cells were employed as representative human CRC cells. We performed the MTT assay and cell cycle analysis to measure apoptotic cell death. The wound healing, Transwell migration, and Matrigel invasion assays were performed to investigate the effect of PT on cell migration/invasion. Western blotting was used to establish the signaling pathway of apoptosis and cell migration/invasion.
RESULTS
PT exerts antiproliferative effect and induces apoptotic cell death of SW620 cells. In addition, PT prevents cell migration and invasion in a dose-dependent manner. Moreover, PT markedly suppressed migration/invasion-related protein expression, including E-cadherin, β-catenin, vimentin, Snail, cyclooxygenase-2, matrix metalloproteinase-2 (MMP-2), and MMP-9 in SW620 cells. PT also inhibited the expression of antiapoptotic proteins (Bcl-2 and Bcl-xL) and activated apoptosis terminal factor (caspase-3) in a dose-dependent manner.
CONCLUSIONS
Our results suggest that PT is a potential novel therapeutic agent for aggressive CRC treatment.

Keyword

Colorectal neoplasms; Parthenolide; Apoptosis; Migration; Invasion

MeSH Terms

Apoptosis
Blotting, Western
Cadherins
Cell Cycle
Cell Death*
Cell Movement
Colorectal Neoplasms
Cyclooxygenase 2
Humans
Matrix Metalloproteinase 2
Snails
Tanacetum parthenium
Vimentin
Wound Healing
Cadherins
Cyclooxygenase 2
Matrix Metalloproteinase 2
Vimentin

Figure

• 

  Fig. 1 Parthenolide (PT) decreases the viability of colorectal cancer cells and induces apoptotic cell death. (A) SW620 cells were treated with PT at concentrations of 0, 5, 10, 20, or 40 µM. After 24 hours of incubation, the MTT assay was conducted to measure viability. Data represent the mean±SE of three independent experiments. (B) Changes in the cell cycle induced by PT treatment. After treatment with PT for 24 hours, SW620 cells were harvested and stained with propidium iodide. The percentage of sub-G1 population is shown in each histogram, and the total number of events analyzed for each condition was 10,000. aP<0.05, bP< 0.01 versus control.

• 

  Fig. 2 Parthenolide (PT) inhibits colorectal cancer cell migration and invasion in vitro. (A) Cell mobility as detected by the wound healing assay. Cells were grown to confluence in 6-well plates, wounded, and treated with the indicated concentrations of PT for 24 hours. Scratch closure was monitored for 24 hours; microscopic images taken at 0 and 24 hours postscratching are shown. Images were captured at a magnification of ×10, and the columns represent the means±SEs of three independent experiments. (B) Migration and invasion assays were performed by the Transwell cell culture system. After incubating for the indicated time, the cells were fixed and stained with crystal violet. Microscopic images were captured at ×20 magnification, and the columns represent the means±SEs of three independent experiments. aP<0.01, bP<0.05 versus the width at 0 hour; cP<0.05, dP<0.01 versus control.

• 

  Fig. 3 Parthenolide (PT) inhibits epithelial-mesenchymal transition-associated markers. Cells were treated with different concentrations of PT, and total cell lysates were extracted by using RIPA buffer. The levels of E-cadherin, β-catenin, vimentin, Snail, matrix metalloproteinase-2 (MMP-2), MMP-9, cyclooxygenase-2 (COX-2), and actin were assessed by Western blotting with the appropriate antibodies. Actin was used as a loading control.

• 

  Fig. 4 Parthenolide (PT) regulates Bcl-2 family members and caspase-3 activation. The cells were treated with different concentrations of PT, and total cell lysates were extracted by using RIPA buffer. The levels of Bcl-2, Bcl-xL, caspase-3, and actin were assessed by Western blotting with the appropriate antibodies. Actin was used as a loading control.

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