Nutr Res Pract.  2017 Apr;11(2):90-96. 10.4162/nrp.2017.11.2.90.

Anti-inflammatory effect of lycopene in SW480 human colorectal cancer cells

• Cha JH
• Kim WK
• Ha AW
• Kim MH
• Chang MJ

• KMID: 2412020
• DOI: http://doi.org/10.4162/nrp.2017.11.2.90

Abstract

BACKGROUND/OBJECTIVES
Although the antioxidative effects of lycopene are generally known, the molecular mechanisms underlying the anti-inflammatory properties of lycopene are not fully elucidated. This study aimed to examine the role and mechanism of lycopene as an inhibitor of inflammation.
METHODS
/MATERIALS: Lipopolysaccharide (LPS)-stimulated SW 480 human colorectal cancer cells were treated with 0, 10, 20, and 30 µM lycopene. The MTT assay was performed to determine the effects of lycopene on cell proliferation. Western blotting was performed to observe the expression of inflammation-related proteins, including nuclear factor-kappa B (NF-κB), inhibitor kappa B (IκB), mitogen-activated protein kinase (MAPK), extracellular signal-related kinase (ERK), c-jun NH2-terminal kinase (JNK), and p38 (p38 MAP kinase). Real-time polymerase chain reaction was performed to investigate the mRNA expression of tumor necrosis factor α (TNF-α), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Concentrations of nitric oxide (NO) and prostaglandin E2 (PGE2) were determined via enzyme-linked immunosorbent assays.
RESULTS
In cells treated with lycopene and LPS, the mRNA expression of TNF-α, IL-1β, IL-6, iNOS, and COX-2 were decreased significantly in a dose-dependent manner (P < 0.05). The concentrations of PGE2 and NO decreased according to the lycopene concentration (P < 0.05). The protein expressions of NF-κB and JNK were decreased significantly according to lycopene concertation (P < 0.05).
CONCLUSIONS
Lycopene restrains NF-κB and JNK activation, which causes inflammation, and suppresses the expression of TNF-α, IL-1β, IL-6, COX-2, and iNOS in SW480 human colorectal cancer cells.

Keyword

Lycopene; colorectal cancer; inflammation; nitric oxide; prostaglandin

MeSH Terms

Blotting, Western
Cell Proliferation
Colorectal Neoplasms*
Cyclooxygenase 2
Dinoprostone
Enzyme-Linked Immunosorbent Assay
Humans*
Inflammation
Interleukin-1beta
Interleukin-6
Nitric Oxide
Nitric Oxide Synthase Type II
Phosphotransferases
Protein Kinases
Real-Time Polymerase Chain Reaction
RNA, Messenger
Tumor Necrosis Factor-alpha
Cyclooxygenase 2
Dinoprostone
Interleukin-1beta
Interleukin-6
Nitric Oxide
Nitric Oxide Synthase Type II
Phosphotransferases
Protein Kinases
RNA, Messenger
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1 Effect of lycopene on cell proliferation in SW480 cells. The effect of lycopene on viable cell numbers were estimated by the MTT assay (A). The effect of lycopene and LPS on viable cell numbers after 24 h incubation the MTT assay (B). Each graph represents the mean ± SE of three independent experiments. Comparison among concentration of lycopene that yielded significant differences at α = 0.05 as indicated by different letters above each graph by Duncan's multiple range test.

• 

  Fig. 2 Effects of lycopene on the mRNA expression of TNF-α, IL-1β and IL-6 in LPS-stimulated SW480 cells. Total RNA was isolated and real-time PCR was performed. Each graph represents the mean ± SE of three independent experiments. Comparison among concentration of lycopene that yielded significant differences at α = 0.05 as indicated by different letters above each graph by Duncan's multiple range test.

• 

  Fig. 3 Effects of lycopene on the mRNA expression of COX-2 (A) and PGE2 concentration (B) in LPS-stimulated SW480 cells. Total RNA was isolated and real-time PCR was performed. PGE2 concentration was measured at 405 nm by ELISA assay. Each graph represents the mean ± SE of three independent experiments. Comparison among concentration of lycopene that yielded significant differences at α = 0.05 as indicated by different letters above each graph by Duncan's multiple range test.

• 

  Fig. 4 Effects of lycopene on the mRNA expression of iNOS (A) and NO (B) concentration in LPS-stimulated SW480 cells. Total RNA was isolated and real-time PCR was performed. NO concentration was measured at 540 nm by Griess reaction assay. Each graph represents the mean ± SE of three independent experiments. Comparison among concentration of lycopene that yielded significant differences at α = 0.05 as indicated by different letters above each graph by Duncan's multiple range test.

• 

  Fig. 5 Effects of lycopene on protein expression of NF-κB (A) and IκB (B) in LPS-stimulated SW480 cells. Protein expression was measured by western blot assay. Actin was used as an internal control in a densitometric analysis. Each graph represents the mean ± SE of three independent experiments. Comparison among concentration of lycopene that yielded significant differences at α = 0.05 as indicated by different letters above each graph by Duncan's multiple range test.

• 

  Fig. 6 Effects of lycopene on protein expression of JNK (A), ERK (B), p38 (C) in LPS-stimulated SW480 cells. Protein expression was measured by western blot assay. Actin was used as an internal control in a densitometric analysis. Each graph represents the mean ± SE of three independent experiments. Comparison among concentration of lycopene that yielded significant differences at α = 0.05 as indicated by different letters above each graph by Duncan's multiple range test.

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