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Ganoderma lucidum polysaccharides in human monocytic leukemia cells: from gene expression to network construction.

Cheng KC, Huang HC, Chen JH, Hsu JW, Cheng HC, Ou CH, Yang WB, Chen ST, Wong CH, Juan HF - BMC Genomics (2007)

Bottom Line: In previous studies, F3, the active component of the polysaccharide extract, was found to activate various cytokines such as IL-1, IL-6, IL-12, and TNF-alpha.The apoptosis induction through the DR3 and DR4/5 death receptors was found to be one of the most significant pathways and play a key role in THP-1 cells after F3 treatment.Our results showed that F3 may induce death receptor ligands to initiate signaling via receptor oligomerization, recruitment of specialized adaptor proteins and activation of caspase cascades.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Life Science, National Taiwan University, Taipei 106, Taiwan. jerekcheng@gmail.com

ABSTRACT

Background: Ganoderma lucidum has been widely used as a herbal medicine for promoting health and longevity in China and other Asian countries. Polysaccharide extracts from Ganoderma lucidum have been reported to exhibit immuno-modulating and anti-tumor activities. In previous studies, F3, the active component of the polysaccharide extract, was found to activate various cytokines such as IL-1, IL-6, IL-12, and TNF-alpha. This gave rise to our investigation on how F3 stimulates immuno-modulating or anti-tumor effects in human leukemia THP-1 cells.

Results: Here, we integrated time-course DNA microarray analysis, quantitative PCR assays, and bioinformatics methods to study the F3-induced effects in THP-1 cells. Significantly disturbed pathways induced by F3 were identified with statistical analysis on microarray data. The apoptosis induction through the DR3 and DR4/5 death receptors was found to be one of the most significant pathways and play a key role in THP-1 cells after F3 treatment. Based on time-course gene expression measurements of the identified pathway, we reconstructed a plausible regulatory network of the involved genes using reverse-engineering computational approach.

Conclusion: Our results showed that F3 may induce death receptor ligands to initiate signaling via receptor oligomerization, recruitment of specialized adaptor proteins and activation of caspase cascades.

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Related in: MedlinePlus

Characterization of F3 induced cell death in human THP-1 cells. Phase-contrast microscopy was used to detect the morphology of the control (A) and F3-treated THP-1 cells (B). Cell shrinkage, shape irregularity, and cellular detachment were observed in F3-treated cells, but not in the control. The control (C) and F3-treated THP-1 cells (D) were stained with 4, 6-diamidino- 2-phenylindole (DAPI). (E) The percentage of chromatin condensed cells. There was apparent difference in cell morphology between the un-treated and F3-treated THP-1 cells.
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Figure 2: Characterization of F3 induced cell death in human THP-1 cells. Phase-contrast microscopy was used to detect the morphology of the control (A) and F3-treated THP-1 cells (B). Cell shrinkage, shape irregularity, and cellular detachment were observed in F3-treated cells, but not in the control. The control (C) and F3-treated THP-1 cells (D) were stained with 4, 6-diamidino- 2-phenylindole (DAPI). (E) The percentage of chromatin condensed cells. There was apparent difference in cell morphology between the un-treated and F3-treated THP-1 cells.

Mentions: After THP-1 cells were treated with F3 (30 μg/mL) for 48 hours, we observed the change of cell morphology under phase-contrast microscope. Differences in cell morphology can be detected between un-treated and F3-treated THP-1 cells. In Figure 2, representative photos of DAPI-staining results are shown. Cell shrinkage, one of cell death characteristics, happened in THP-1 cells after 48 hours treatment with F3 (Figure 2A and 2B). During cell apoptosis, an early event is the nuclear chromatin condensation, leading to the degradation of genomic DNA. DAPI nuclear staining was performed to check the apoptotic changes shown by cell morphology (Figure 2C and 2D). The percentage of chromatin condensed cells in F3-treated culture saw a significant increase (Figure 2E). Shrunken nucleus and apoptotic bodies in DAPI staining were features in determining whether cells had undergone apoptosis. These results indicated that incubation of THP-1 cells with F3 for 48 hours would lead to cell aggregations and apoptosis.


Ganoderma lucidum polysaccharides in human monocytic leukemia cells: from gene expression to network construction.

Cheng KC, Huang HC, Chen JH, Hsu JW, Cheng HC, Ou CH, Yang WB, Chen ST, Wong CH, Juan HF - BMC Genomics (2007)

Characterization of F3 induced cell death in human THP-1 cells. Phase-contrast microscopy was used to detect the morphology of the control (A) and F3-treated THP-1 cells (B). Cell shrinkage, shape irregularity, and cellular detachment were observed in F3-treated cells, but not in the control. The control (C) and F3-treated THP-1 cells (D) were stained with 4, 6-diamidino- 2-phenylindole (DAPI). (E) The percentage of chromatin condensed cells. There was apparent difference in cell morphology between the un-treated and F3-treated THP-1 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2211495&req=5

Figure 2: Characterization of F3 induced cell death in human THP-1 cells. Phase-contrast microscopy was used to detect the morphology of the control (A) and F3-treated THP-1 cells (B). Cell shrinkage, shape irregularity, and cellular detachment were observed in F3-treated cells, but not in the control. The control (C) and F3-treated THP-1 cells (D) were stained with 4, 6-diamidino- 2-phenylindole (DAPI). (E) The percentage of chromatin condensed cells. There was apparent difference in cell morphology between the un-treated and F3-treated THP-1 cells.
Mentions: After THP-1 cells were treated with F3 (30 μg/mL) for 48 hours, we observed the change of cell morphology under phase-contrast microscope. Differences in cell morphology can be detected between un-treated and F3-treated THP-1 cells. In Figure 2, representative photos of DAPI-staining results are shown. Cell shrinkage, one of cell death characteristics, happened in THP-1 cells after 48 hours treatment with F3 (Figure 2A and 2B). During cell apoptosis, an early event is the nuclear chromatin condensation, leading to the degradation of genomic DNA. DAPI nuclear staining was performed to check the apoptotic changes shown by cell morphology (Figure 2C and 2D). The percentage of chromatin condensed cells in F3-treated culture saw a significant increase (Figure 2E). Shrunken nucleus and apoptotic bodies in DAPI staining were features in determining whether cells had undergone apoptosis. These results indicated that incubation of THP-1 cells with F3 for 48 hours would lead to cell aggregations and apoptosis.

Bottom Line: In previous studies, F3, the active component of the polysaccharide extract, was found to activate various cytokines such as IL-1, IL-6, IL-12, and TNF-alpha.The apoptosis induction through the DR3 and DR4/5 death receptors was found to be one of the most significant pathways and play a key role in THP-1 cells after F3 treatment.Our results showed that F3 may induce death receptor ligands to initiate signaling via receptor oligomerization, recruitment of specialized adaptor proteins and activation of caspase cascades.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Life Science, National Taiwan University, Taipei 106, Taiwan. jerekcheng@gmail.com

ABSTRACT

Background: Ganoderma lucidum has been widely used as a herbal medicine for promoting health and longevity in China and other Asian countries. Polysaccharide extracts from Ganoderma lucidum have been reported to exhibit immuno-modulating and anti-tumor activities. In previous studies, F3, the active component of the polysaccharide extract, was found to activate various cytokines such as IL-1, IL-6, IL-12, and TNF-alpha. This gave rise to our investigation on how F3 stimulates immuno-modulating or anti-tumor effects in human leukemia THP-1 cells.

Results: Here, we integrated time-course DNA microarray analysis, quantitative PCR assays, and bioinformatics methods to study the F3-induced effects in THP-1 cells. Significantly disturbed pathways induced by F3 were identified with statistical analysis on microarray data. The apoptosis induction through the DR3 and DR4/5 death receptors was found to be one of the most significant pathways and play a key role in THP-1 cells after F3 treatment. Based on time-course gene expression measurements of the identified pathway, we reconstructed a plausible regulatory network of the involved genes using reverse-engineering computational approach.

Conclusion: Our results showed that F3 may induce death receptor ligands to initiate signaling via receptor oligomerization, recruitment of specialized adaptor proteins and activation of caspase cascades.

Show MeSH
Related in: MedlinePlus