Nutr Res Pract.  2009 Sep;3(3):180-184.

beta-Glucan enhanced apoptosis in human colon cancer cells SNU-C4

• Kim MJ
• Hong SY
• Kim SK
• Cheong C
• Park HJ
• Chun HK
• Jang KH
• Yoon BD
• Kim CH
• Kang SA

Abstract

The apoptotic effect of bacteria-derived beta-glucan was investigated in human colon cancer cells SNU-C4 using terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, reverse transcription-polymerase chain reaction (RT-PCR) expressions of Bcl-2, Bax, and Caspase-3 genes, and assay of caspase-3 enzyme activity. beta-Glucan of 10, 50, and 100 microg/mL decreased cell viability in a dose-dependent manner with typical apoptotic characteristics, such as morphological changes of chromatin condensation and apoptotic body formation from TUNEL assay. In addition, beta-glucan (100 microgram/mL) decreased the expression of Bcl-2 by 0.6 times, whereas the expression of Bax and Caspase-3 were increased by 3.1 and 2.3 times, respectively, compared to untreated control group. Furthermore, the caspase-3 activity in the beta-glucan-treated group was significantly increased compared to those in control group (P < 0.05). Bacterial derived beta-glucan could be used as an effective compound inducing apoptosis in human colon cancer.

Keyword

beta-glucan; apoptosis; colon cancer; caspase-3

MeSH Terms

Apoptosis
Caspase 3
Cell Survival
Chromatin
Colon
Colonic Neoplasms
DNA Nucleotidylexotransferase
Humans
In Situ Nick-End Labeling
Caspase 3
Chromatin
DNA Nucleotidylexotransferase

Figure

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  Fig. 1 Cell viability of β-glucan in human colon cancer cells SNU-C4 for 24 hrs. Viability was determined with MTT assay. Results are presented as mean ± SEM. The experiments were done triplicate. Different letters denote significant differences within each group (P < 0.05).

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  Fig. 2 Morphological change of β-glucan induced cell death in human colon cancer cells SNU-C4. Cells were cultured without resveratrol (A) or with 10, 50, and 100 µg/mL of β-glucan (B, C, and D). Morphology (top): phase-contrast microscopy showed cell shrinkage, irregularity of shape in β-glucan treated cultures. TUNEL assay (bottom): SNU-668 cells stained using TUNEL method. Condensed and marginated chromatin showed to be stained dark brown. The experiments were done triplicate. Scale bar, 100 µm.

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  Fig. 3 RT-PCR analysis of Bcl-2, Bax and Caspase-3. As the internal control, Cyclophilin mRNA was also β-glucan transcribed. A: control, B: 10 µg/mL of β-glucan treated group, C: 50 µg/mL of β-glucan treated group, D: 100 µg/mL of β-glucan treated group. The experiments were done triplicate. Different letters denote significant differences within each group (P < 0.05).

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  Fig. 4 Caspase-3 activity of β-glucan treated SNU-C4. The rate of DEVD-pNA cleavage was measured at 405 nm. Caspase-3 is a positive control. The experiments were done triplicate. Different letters means significantly different among groups (P < 0.05).

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