Limits...
Gene Expression Profiling and Pathway Network Analysis Predicts a Novel Antitumor Function for a Botanical-Derived Drug, PG2.

Kuo YL, Chen CH, Chuang TH, Hua WK, Lin WJ, Hsu WH, Chang PM, Hsu SL, Huang TH, Kao CY, Huang CY - Evid Based Complement Alternat Med (2015)

Bottom Line: The results showed that PG2 product batches were consistent and of high quality.Within the PG2 signature, there were five genes associated with doxorubicin: IL-8, MDM4, BCL2, PRODH2, and BIRC5.Moreover, the combination of PG2 and doxorubicin had a synergistic effect on induced cell death in HL-60 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Computer Science and Information Engineering, National Taiwan University, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan.

ABSTRACT
PG2 is a botanical drug that is mostly composed of Astragalus polysaccharides (APS). Its role in hematopoiesis and relieving cancer-related fatigue has recently been clinically investigated in cancer patients. However, systematic analyses of its functions are still limited. The aim of this study was to use microarray-based expression profiling to evaluate the quality and consistency of PG2 from three different product batches and to study biological mechanisms of PG2. An integrative molecular analysis approach has been designed to examine significant PG2-induced signatures in HL-60 leukemia cells. A quantitative analysis of gene expression signatures was conducted for PG2 by hierarchical clustering of correlation coefficients. The results showed that PG2 product batches were consistent and of high quality. These batches were also functionally equivalent to each other with regard to how they modulated the immune and hematopoietic systems. Within the PG2 signature, there were five genes associated with doxorubicin: IL-8, MDM4, BCL2, PRODH2, and BIRC5. Moreover, the combination of PG2 and doxorubicin had a synergistic effect on induced cell death in HL-60 cells. Together with the bioinformatics-based approach, gene expression profiling provided a quantitative measurement for the quality and consistency of herbal medicines and revealed new roles (e.g., immune modulation) for PG2 in cancer treatment.

No MeSH data available.


Related in: MedlinePlus

Network analysis of PG2-enriched signature reveals the connection to immune function. (a) The subnetwork was composed of 87 genes, including graph-based clique and overexpressed genes. These genes were subsequently analyzed using CPDB pathway analysis. As indicated in this figure, several subnetworks were identified, including several immune-related signaling pathways, such as interleukin-mediated signaling, toll-like signaling, Trk receptor signaling, and the CTLA4 inhibitory pathway. Links represent a protein-protein interaction in the network. The size of each node is based on the number of links to other nodes. Nodes with upregulated and downregulated genes are colored in red and green, respectively. The depth of color indicates the gene expression level fold change. Several genes from PG2-enriched signature, represented by a black borderline on a node (e.g., STAT5A in CTLA4 inhibitory pathway), are not in the original pathway analysis but are included in the pathways of the subnetworks based on a literature survey. The network was visualized using Cytoscape 3.0 (http://www.cytoscape.org/). HL-60 cells were treated with different concentrations of PG2 for 6 hr. (b) PTPN11 and (c) NFKB2 mRNA was measured by Q-RT PCR and normalized to β-actin (n = 3). PG2 treatment alone drastically increased the production of IL-1β (d) and IL-6 (e) in human THP-1 macrophages (n = 3) (#P < 0.05; ##P < 0.01; ###P < 0.001).
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4417974&req=5

fig3: Network analysis of PG2-enriched signature reveals the connection to immune function. (a) The subnetwork was composed of 87 genes, including graph-based clique and overexpressed genes. These genes were subsequently analyzed using CPDB pathway analysis. As indicated in this figure, several subnetworks were identified, including several immune-related signaling pathways, such as interleukin-mediated signaling, toll-like signaling, Trk receptor signaling, and the CTLA4 inhibitory pathway. Links represent a protein-protein interaction in the network. The size of each node is based on the number of links to other nodes. Nodes with upregulated and downregulated genes are colored in red and green, respectively. The depth of color indicates the gene expression level fold change. Several genes from PG2-enriched signature, represented by a black borderline on a node (e.g., STAT5A in CTLA4 inhibitory pathway), are not in the original pathway analysis but are included in the pathways of the subnetworks based on a literature survey. The network was visualized using Cytoscape 3.0 (http://www.cytoscape.org/). HL-60 cells were treated with different concentrations of PG2 for 6 hr. (b) PTPN11 and (c) NFKB2 mRNA was measured by Q-RT PCR and normalized to β-actin (n = 3). PG2 treatment alone drastically increased the production of IL-1β (d) and IL-6 (e) in human THP-1 macrophages (n = 3) (#P < 0.05; ##P < 0.01; ###P < 0.001).

Mentions: Using the PG2-enriched signature to construct a PPI subnetwork, as shown in the center of Figure 3(a), six crucial immune response pathways were identified to be significant (P value < 0.05 and q-value < 0.05). These pathways included natural killer (NK) cell-mediated cytotoxicity, signaling by the B cell receptor, neurotrophic factor-mediated Trk receptor signaling, toll-like receptor cascades, the CTLA4 inhibitory pathway, and interleukin-mediated signaling (Figure 3(a)). Briefly, NK cells produce inflammatory cytokines and interact with other cells of the innate and adaptive immune system. NK cells also have antitumor functions as they efficiently attack target cancer cells that express low levels of MHC class I molecules [23]. Based on previous reports, α-galactosylceramide analogs, which are used as immunotherapy drugs, can induce IL-2 secretion to enhance the cytotoxic effects of NK cells [24, 25]. Importantly, activation of B lymphocytes (referred to as B cells) plays an important role in an efficient antitumor response by the immune system. Interestingly, some cytokines, such as TNF-α, IL-2, and TGF-β, can regulate synaptic plasticity and influence CNS functions. Microglia, which are immune cells found in the brain and spinal cord, can be detected in CNS injuries and have been found to produce and secrete a variety of neurotrophic factors, such as NGF, BDNF, neurotrophin-3 (NT-3), and neurotrophin-4/5 (NT-4/5). Microglia express members of the tyrosine kinase (Trk) receptor family, such as TrkA, TrkB, and TrkC [26]. Moreover, previous reports have shown that NT-3 increases microglial proliferation and phagocytosis through Trk receptor expression and stimulates microglial cells to attack cancer cells [27].


Gene Expression Profiling and Pathway Network Analysis Predicts a Novel Antitumor Function for a Botanical-Derived Drug, PG2.

Kuo YL, Chen CH, Chuang TH, Hua WK, Lin WJ, Hsu WH, Chang PM, Hsu SL, Huang TH, Kao CY, Huang CY - Evid Based Complement Alternat Med (2015)

Network analysis of PG2-enriched signature reveals the connection to immune function. (a) The subnetwork was composed of 87 genes, including graph-based clique and overexpressed genes. These genes were subsequently analyzed using CPDB pathway analysis. As indicated in this figure, several subnetworks were identified, including several immune-related signaling pathways, such as interleukin-mediated signaling, toll-like signaling, Trk receptor signaling, and the CTLA4 inhibitory pathway. Links represent a protein-protein interaction in the network. The size of each node is based on the number of links to other nodes. Nodes with upregulated and downregulated genes are colored in red and green, respectively. The depth of color indicates the gene expression level fold change. Several genes from PG2-enriched signature, represented by a black borderline on a node (e.g., STAT5A in CTLA4 inhibitory pathway), are not in the original pathway analysis but are included in the pathways of the subnetworks based on a literature survey. The network was visualized using Cytoscape 3.0 (http://www.cytoscape.org/). HL-60 cells were treated with different concentrations of PG2 for 6 hr. (b) PTPN11 and (c) NFKB2 mRNA was measured by Q-RT PCR and normalized to β-actin (n = 3). PG2 treatment alone drastically increased the production of IL-1β (d) and IL-6 (e) in human THP-1 macrophages (n = 3) (#P < 0.05; ##P < 0.01; ###P < 0.001).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4417974&req=5

fig3: Network analysis of PG2-enriched signature reveals the connection to immune function. (a) The subnetwork was composed of 87 genes, including graph-based clique and overexpressed genes. These genes were subsequently analyzed using CPDB pathway analysis. As indicated in this figure, several subnetworks were identified, including several immune-related signaling pathways, such as interleukin-mediated signaling, toll-like signaling, Trk receptor signaling, and the CTLA4 inhibitory pathway. Links represent a protein-protein interaction in the network. The size of each node is based on the number of links to other nodes. Nodes with upregulated and downregulated genes are colored in red and green, respectively. The depth of color indicates the gene expression level fold change. Several genes from PG2-enriched signature, represented by a black borderline on a node (e.g., STAT5A in CTLA4 inhibitory pathway), are not in the original pathway analysis but are included in the pathways of the subnetworks based on a literature survey. The network was visualized using Cytoscape 3.0 (http://www.cytoscape.org/). HL-60 cells were treated with different concentrations of PG2 for 6 hr. (b) PTPN11 and (c) NFKB2 mRNA was measured by Q-RT PCR and normalized to β-actin (n = 3). PG2 treatment alone drastically increased the production of IL-1β (d) and IL-6 (e) in human THP-1 macrophages (n = 3) (#P < 0.05; ##P < 0.01; ###P < 0.001).
Mentions: Using the PG2-enriched signature to construct a PPI subnetwork, as shown in the center of Figure 3(a), six crucial immune response pathways were identified to be significant (P value < 0.05 and q-value < 0.05). These pathways included natural killer (NK) cell-mediated cytotoxicity, signaling by the B cell receptor, neurotrophic factor-mediated Trk receptor signaling, toll-like receptor cascades, the CTLA4 inhibitory pathway, and interleukin-mediated signaling (Figure 3(a)). Briefly, NK cells produce inflammatory cytokines and interact with other cells of the innate and adaptive immune system. NK cells also have antitumor functions as they efficiently attack target cancer cells that express low levels of MHC class I molecules [23]. Based on previous reports, α-galactosylceramide analogs, which are used as immunotherapy drugs, can induce IL-2 secretion to enhance the cytotoxic effects of NK cells [24, 25]. Importantly, activation of B lymphocytes (referred to as B cells) plays an important role in an efficient antitumor response by the immune system. Interestingly, some cytokines, such as TNF-α, IL-2, and TGF-β, can regulate synaptic plasticity and influence CNS functions. Microglia, which are immune cells found in the brain and spinal cord, can be detected in CNS injuries and have been found to produce and secrete a variety of neurotrophic factors, such as NGF, BDNF, neurotrophin-3 (NT-3), and neurotrophin-4/5 (NT-4/5). Microglia express members of the tyrosine kinase (Trk) receptor family, such as TrkA, TrkB, and TrkC [26]. Moreover, previous reports have shown that NT-3 increases microglial proliferation and phagocytosis through Trk receptor expression and stimulates microglial cells to attack cancer cells [27].

Bottom Line: The results showed that PG2 product batches were consistent and of high quality.Within the PG2 signature, there were five genes associated with doxorubicin: IL-8, MDM4, BCL2, PRODH2, and BIRC5.Moreover, the combination of PG2 and doxorubicin had a synergistic effect on induced cell death in HL-60 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Computer Science and Information Engineering, National Taiwan University, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan.

ABSTRACT
PG2 is a botanical drug that is mostly composed of Astragalus polysaccharides (APS). Its role in hematopoiesis and relieving cancer-related fatigue has recently been clinically investigated in cancer patients. However, systematic analyses of its functions are still limited. The aim of this study was to use microarray-based expression profiling to evaluate the quality and consistency of PG2 from three different product batches and to study biological mechanisms of PG2. An integrative molecular analysis approach has been designed to examine significant PG2-induced signatures in HL-60 leukemia cells. A quantitative analysis of gene expression signatures was conducted for PG2 by hierarchical clustering of correlation coefficients. The results showed that PG2 product batches were consistent and of high quality. These batches were also functionally equivalent to each other with regard to how they modulated the immune and hematopoietic systems. Within the PG2 signature, there were five genes associated with doxorubicin: IL-8, MDM4, BCL2, PRODH2, and BIRC5. Moreover, the combination of PG2 and doxorubicin had a synergistic effect on induced cell death in HL-60 cells. Together with the bioinformatics-based approach, gene expression profiling provided a quantitative measurement for the quality and consistency of herbal medicines and revealed new roles (e.g., immune modulation) for PG2 in cancer treatment.

No MeSH data available.


Related in: MedlinePlus