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Identification of Target Genes Involved in Wound Healing Angiogenesis of Endothelial Cells with the Treatment of a Chinese 2-Herb Formula.

Tam JC, Ko CH, Koon CM, Cheng Z, Lok WH, Lau CP, Leung PC, Fung KP, Chan WY, Lau CB - PLoS ONE (2015)

Bottom Line: We had previously demonstrated that a Chinese 2-herb formula (NF3) significantly stimulated angiogenesis of HUVEC in wound healing.The microarray results illustrated that different panels of differentially expressed genes were strictly governed in NF3-treated HUVEC in a time-regulated manner.This study provided concrete scientific evidence in support of the regulatory role of NF3 on endothelial cells involved in wound healing angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong; State Key Laboratory of Phytochemistry and Plant Resources in West China, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.

ABSTRACT
Angiogenesis is vitally important in diabetic wound healing. We had previously demonstrated that a Chinese 2-herb formula (NF3) significantly stimulated angiogenesis of HUVEC in wound healing. However, the molecular mechanism has not yet been elucidated. In line with this, global expression profiling of NF3-treated HUVEC was performed so as to assess the regulatory role of NF3 involved in the underlying signaling pathways in wound healing angiogenesis. The microarray results illustrated that different panels of differentially expressed genes were strictly governed in NF3-treated HUVEC in a time-regulated manner. The microarray analysis followed by qRT-PCR and western blotting verification of NF3-treated HUVEC at 6 h revealed the involvement of various genes in diverse biological process, e.g., MAP3K14 in anti-inflammation; SLC5A8 in anti-tumorogenesis; DNAJB7 in protein translation; BIRC5, EPCAM, INSL4, MMP8 and NPR3 in cell proliferation; CXCR7, EPCAM, HAND1 and MMP8 in migration; CXCR7, EPCAM and MMP8 in tubular formation; and BIRC5, CXCR7, EPCAM, HAND1, MMP8 and UBD in angiogenesis. After 16 h incubation of NF3, other sets of genes were shown with differential expression in HUVEC, e.g., IL1RAPL2 and NR1H4 in anti-inflammation; miR28 in anti-tumorogenesis; GRIN1 and LCN1 in anti-oxidation; EPB41 in intracellular signal transduction; PRL and TFAP2A in cell proliferation; miR28, PRL and SCG2 in cell migration; PRL in tubular formation; and miR28, NR1H4 and PRL in angiogenesis. This study provided concrete scientific evidence in support of the regulatory role of NF3 on endothelial cells involved in wound healing angiogenesis.

No MeSH data available.


Related in: MedlinePlus

Network connectivity of identified genes in NF3-treated HUVEC by GePS.(A) Differentially expressed genes with frequent co-citations and interactions in NF3 treated HUVEC at 6 h. (B) Differentially expressed genes with frequent co-citations and interactions in NF3 treated HUVEC at 16 h. Genes were identified by their gene symbols (S1 and S2 Tables). Dashed lines indicated association by co-citation. Up arrow and down arrow corresponded to up- or down-regulation, respectively.
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pone.0139342.g003: Network connectivity of identified genes in NF3-treated HUVEC by GePS.(A) Differentially expressed genes with frequent co-citations and interactions in NF3 treated HUVEC at 6 h. (B) Differentially expressed genes with frequent co-citations and interactions in NF3 treated HUVEC at 16 h. Genes were identified by their gene symbols (S1 and S2 Tables). Dashed lines indicated association by co-citation. Up arrow and down arrow corresponded to up- or down-regulation, respectively.

Mentions: To elucidate the regulatory mechanisms of NF3 on the differentially expressed genes of HUVEC involved in wound healing angiogenesis, the genes were annotated by GePS according to their relative interactions and frequent co-citation from data and literature mining so as to predict their biological processes and underlying signaling pathways. The connectivity map organized genes based on their interactions, which could further extend the input genes by transcription factors or transcriptional downstream targets. In Fig 3, genes with differential expression in HUVEC after 6 h and 16 h of NF3 treatment were linked in the connectivity network. The advantage of connectivity map by GePS was to sort out genes with interactions for subsequent pathway prediction. All the target genes were further analyzed using the Universal Protein Resource (UniProt) in order to categorize them according to gene ontology (GO) and respective protein families. As illustrated in Table 3 of HUVEC after 6 h of NF3 treatment, BIRC5 was annotated to regulate cell cycle and migration behavior (microtubule organizing center). CXCR7 and HAND1 were responsible for neovascularization of endothelial cells. Some genes (DNAJB7 and HAND1) possessed DNA binding transcription factor activity. EPCAM, GUCY2C and INSL4 were directly involved in cell proliferation. Genes with protease activity participating in different cellular reaction included MAP3K14, MMP8 and PGC. On the other hand, after 16 h of NF3 treatment (Table 4), different genes were triggered and expressed in HUVEC. EPB41 was identified to participate in actin cytoskeleton organization, governing cell motility. IL1RAPL2 contained interleukin-1 receptor activity, transmitting inflammatory signal of interleukin-1. SCG2 was the candidate gene responsible for angiogenesis as well as regulation of proliferation and migration of endothelial cells. NR1H4, TFAP2A and ZNF711 belonged to DNA binding transcription factor activity or regulation of transcription. From the GO analysis, the differentially expressed genes in HUVEC would provide insight of the regulatory mechanisms as well as the prediction of the signaling pathways elicited by NF3 on mature endothelial cells in modulation of wound healing angiogenesis.


Identification of Target Genes Involved in Wound Healing Angiogenesis of Endothelial Cells with the Treatment of a Chinese 2-Herb Formula.

Tam JC, Ko CH, Koon CM, Cheng Z, Lok WH, Lau CP, Leung PC, Fung KP, Chan WY, Lau CB - PLoS ONE (2015)

Network connectivity of identified genes in NF3-treated HUVEC by GePS.(A) Differentially expressed genes with frequent co-citations and interactions in NF3 treated HUVEC at 6 h. (B) Differentially expressed genes with frequent co-citations and interactions in NF3 treated HUVEC at 16 h. Genes were identified by their gene symbols (S1 and S2 Tables). Dashed lines indicated association by co-citation. Up arrow and down arrow corresponded to up- or down-regulation, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591983&req=5

pone.0139342.g003: Network connectivity of identified genes in NF3-treated HUVEC by GePS.(A) Differentially expressed genes with frequent co-citations and interactions in NF3 treated HUVEC at 6 h. (B) Differentially expressed genes with frequent co-citations and interactions in NF3 treated HUVEC at 16 h. Genes were identified by their gene symbols (S1 and S2 Tables). Dashed lines indicated association by co-citation. Up arrow and down arrow corresponded to up- or down-regulation, respectively.
Mentions: To elucidate the regulatory mechanisms of NF3 on the differentially expressed genes of HUVEC involved in wound healing angiogenesis, the genes were annotated by GePS according to their relative interactions and frequent co-citation from data and literature mining so as to predict their biological processes and underlying signaling pathways. The connectivity map organized genes based on their interactions, which could further extend the input genes by transcription factors or transcriptional downstream targets. In Fig 3, genes with differential expression in HUVEC after 6 h and 16 h of NF3 treatment were linked in the connectivity network. The advantage of connectivity map by GePS was to sort out genes with interactions for subsequent pathway prediction. All the target genes were further analyzed using the Universal Protein Resource (UniProt) in order to categorize them according to gene ontology (GO) and respective protein families. As illustrated in Table 3 of HUVEC after 6 h of NF3 treatment, BIRC5 was annotated to regulate cell cycle and migration behavior (microtubule organizing center). CXCR7 and HAND1 were responsible for neovascularization of endothelial cells. Some genes (DNAJB7 and HAND1) possessed DNA binding transcription factor activity. EPCAM, GUCY2C and INSL4 were directly involved in cell proliferation. Genes with protease activity participating in different cellular reaction included MAP3K14, MMP8 and PGC. On the other hand, after 16 h of NF3 treatment (Table 4), different genes were triggered and expressed in HUVEC. EPB41 was identified to participate in actin cytoskeleton organization, governing cell motility. IL1RAPL2 contained interleukin-1 receptor activity, transmitting inflammatory signal of interleukin-1. SCG2 was the candidate gene responsible for angiogenesis as well as regulation of proliferation and migration of endothelial cells. NR1H4, TFAP2A and ZNF711 belonged to DNA binding transcription factor activity or regulation of transcription. From the GO analysis, the differentially expressed genes in HUVEC would provide insight of the regulatory mechanisms as well as the prediction of the signaling pathways elicited by NF3 on mature endothelial cells in modulation of wound healing angiogenesis.

Bottom Line: We had previously demonstrated that a Chinese 2-herb formula (NF3) significantly stimulated angiogenesis of HUVEC in wound healing.The microarray results illustrated that different panels of differentially expressed genes were strictly governed in NF3-treated HUVEC in a time-regulated manner.This study provided concrete scientific evidence in support of the regulatory role of NF3 on endothelial cells involved in wound healing angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong; State Key Laboratory of Phytochemistry and Plant Resources in West China, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.

ABSTRACT
Angiogenesis is vitally important in diabetic wound healing. We had previously demonstrated that a Chinese 2-herb formula (NF3) significantly stimulated angiogenesis of HUVEC in wound healing. However, the molecular mechanism has not yet been elucidated. In line with this, global expression profiling of NF3-treated HUVEC was performed so as to assess the regulatory role of NF3 involved in the underlying signaling pathways in wound healing angiogenesis. The microarray results illustrated that different panels of differentially expressed genes were strictly governed in NF3-treated HUVEC in a time-regulated manner. The microarray analysis followed by qRT-PCR and western blotting verification of NF3-treated HUVEC at 6 h revealed the involvement of various genes in diverse biological process, e.g., MAP3K14 in anti-inflammation; SLC5A8 in anti-tumorogenesis; DNAJB7 in protein translation; BIRC5, EPCAM, INSL4, MMP8 and NPR3 in cell proliferation; CXCR7, EPCAM, HAND1 and MMP8 in migration; CXCR7, EPCAM and MMP8 in tubular formation; and BIRC5, CXCR7, EPCAM, HAND1, MMP8 and UBD in angiogenesis. After 16 h incubation of NF3, other sets of genes were shown with differential expression in HUVEC, e.g., IL1RAPL2 and NR1H4 in anti-inflammation; miR28 in anti-tumorogenesis; GRIN1 and LCN1 in anti-oxidation; EPB41 in intracellular signal transduction; PRL and TFAP2A in cell proliferation; miR28, PRL and SCG2 in cell migration; PRL in tubular formation; and miR28, NR1H4 and PRL in angiogenesis. This study provided concrete scientific evidence in support of the regulatory role of NF3 on endothelial cells involved in wound healing angiogenesis.

No MeSH data available.


Related in: MedlinePlus