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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

Comparisons between oligonucelotide microarray and Q-PCR results. (A) THP-1 cells were treated with F3 for 6 hours (A) and 24 hours (B) and with LPS for 24 hours (C). The gene expressions showed consistent trends between the microarray and Q-PCR results. The error bar came from n > 3.
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Figure 9: Comparisons between oligonucelotide microarray and Q-PCR results. (A) THP-1 cells were treated with F3 for 6 hours (A) and 24 hours (B) and with LPS for 24 hours (C). The gene expressions showed consistent trends between the microarray and Q-PCR results. The error bar came from n > 3.

Mentions: To further validate our findings from the microarray analysis, we selected a set of genes known for their involvement in apoptosis through death receptors, and carried out gene expression studies using Q-PCR. 18 gene expressions related to the death receptor pathway were examined between F3- or LPS-treated THP-1 cells for 3, 6, 12, and 24 hours. mRNAs were reverse-transcribed and amplified through Q-PCR using primers specific for each gene of interest; the housekeeping gene, GAPDH, was used as internal control. Each experiment was repeated three times. The results are shown in Figure 7 and 8. Significance analysis of Q-PCR measurements was performed by EDGE software package [54]. There were nine significantly differential gene expressions in F3-treated THP-1 cells, including TNFRSF10B, CASP7, CASP6, TRADD, CASP3, TNFSF12, baculoviral IAP repeat-containing 2 (BIRC2/c-IAP), conserved helix-loop-helix ubiquitous kinase (CHUK/IKKα), and NF-κB (NFKB1); whereas in LPS-treated THP-1 cells these were TNFRSF12, CASP7, TNFSF12, CASP6, TRAF2, and NFKB1. Figure 9 shows that nearly all genes are consistent between the microarray and Q-PCR data.


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)

Comparisons between oligonucelotide microarray and Q-PCR results. (A) THP-1 cells were treated with F3 for 6 hours (A) and 24 hours (B) and with LPS for 24 hours (C). The gene expressions showed consistent trends between the microarray and Q-PCR results. The error bar came from n > 3.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 9: Comparisons between oligonucelotide microarray and Q-PCR results. (A) THP-1 cells were treated with F3 for 6 hours (A) and 24 hours (B) and with LPS for 24 hours (C). The gene expressions showed consistent trends between the microarray and Q-PCR results. The error bar came from n > 3.
Mentions: To further validate our findings from the microarray analysis, we selected a set of genes known for their involvement in apoptosis through death receptors, and carried out gene expression studies using Q-PCR. 18 gene expressions related to the death receptor pathway were examined between F3- or LPS-treated THP-1 cells for 3, 6, 12, and 24 hours. mRNAs were reverse-transcribed and amplified through Q-PCR using primers specific for each gene of interest; the housekeeping gene, GAPDH, was used as internal control. Each experiment was repeated three times. The results are shown in Figure 7 and 8. Significance analysis of Q-PCR measurements was performed by EDGE software package [54]. There were nine significantly differential gene expressions in F3-treated THP-1 cells, including TNFRSF10B, CASP7, CASP6, TRADD, CASP3, TNFSF12, baculoviral IAP repeat-containing 2 (BIRC2/c-IAP), conserved helix-loop-helix ubiquitous kinase (CHUK/IKKα), and NF-κB (NFKB1); whereas in LPS-treated THP-1 cells these were TNFRSF12, CASP7, TNFSF12, CASP6, TRAF2, and NFKB1. Figure 9 shows that nearly all genes are consistent between the microarray and Q-PCR data.

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