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Whole Genome Expression Profiling and Signal Pathway Screening of MSCs in Ankylosing Spondylitis.

Li Y, Wang P, Xie Z, Huang L, Yang R, Gao L, Tang Y, Zhang X, Ye J, Chen K, Cai Z, Wu Y, Shen H - Stem Cells Int (2014)

Bottom Line: Our results showed that in normal media 676 genes were differentially expressed in AS, 354 upregulated and 322 downregulated, while in an inflammatory environment 1767 genes were differentially expressed in AS, 1230 upregulated and 537 downregulated.In addition, by KEGG pathway analysis, 14 key genes from the MAPK signaling and 8 key genes from the TLR signaling pathway were identified as differentially regulated.The results of qRT-PCR verified the expression variation of the 9 genes mentioned above.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, No. 107 Yanjiang Road West, Guangzhou 510120, China.

ABSTRACT
The pathogenesis of dysfunctional immunoregulation of mesenchymal stem cells (MSCs) in ankylosing spondylitis (AS) is thought to be a complex process that involves multiple genetic alterations. In this study, MSCs derived from both healthy donors and AS patients were cultured in normal media or media mimicking an inflammatory environment. Whole genome expression profiling analysis of 33,351 genes was performed and differentially expressed genes related to AS were analyzed by GO term analysis and KEGG pathway analysis. Our results showed that in normal media 676 genes were differentially expressed in AS, 354 upregulated and 322 downregulated, while in an inflammatory environment 1767 genes were differentially expressed in AS, 1230 upregulated and 537 downregulated. GO analysis showed that these genes were mainly related to cellular processes, physiological processes, biological regulation, regulation of biological processes, and binding. In addition, by KEGG pathway analysis, 14 key genes from the MAPK signaling and 8 key genes from the TLR signaling pathway were identified as differentially regulated. The results of qRT-PCR verified the expression variation of the 9 genes mentioned above. Our study found that in an inflammatory environment ankylosing spondylitis pathogenesis may be related to activation of the MAPK and TLR signaling pathways.

No MeSH data available.


Related in: MedlinePlus

Verification of differentially expressed genes of MSCs in AS patients cultured in inflammatory environment. RNA was isolated from MSCs of HD (n = 5) and AS patients (n = 5) after MSCs were cultured in inflammatory environment mimicked by TNF-α (10 ng/mL) and IFN-γ (10 ng/mL). Comparison of the levels of differentially expressed genes in MSCs from both groups was based on quantitative real-time polymerase chain reaction (qRT-PCR). Beta-actin was used as an internal control. The results showed that when grown in inflammatory culture medium, 9 of 22 differentially expressed genes from whole genomic expression analysis in AS patients were truly differentially expressed. These genes include GNA12, DUSP1, MAPK11, RAC1, MAPK3, TRAF3, IFNAR1, IL6, and IFNA1. The symbol “∗” represents P < 0.05.
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fig3: Verification of differentially expressed genes of MSCs in AS patients cultured in inflammatory environment. RNA was isolated from MSCs of HD (n = 5) and AS patients (n = 5) after MSCs were cultured in inflammatory environment mimicked by TNF-α (10 ng/mL) and IFN-γ (10 ng/mL). Comparison of the levels of differentially expressed genes in MSCs from both groups was based on quantitative real-time polymerase chain reaction (qRT-PCR). Beta-actin was used as an internal control. The results showed that when grown in inflammatory culture medium, 9 of 22 differentially expressed genes from whole genomic expression analysis in AS patients were truly differentially expressed. These genes include GNA12, DUSP1, MAPK11, RAC1, MAPK3, TRAF3, IFNAR1, IL6, and IFNA1. The symbol “∗” represents P < 0.05.

Mentions: KEGG pathway analysis was used to discover key signaling pathways and relationships between differentially expressed genes. In the inflammatory environment, the two highest scores of indicators in the AS group were hsa04010: mitogen-activated protein kinase (MAPK) signaling pathway, and hsa04620: toll-like receptor (TLR) signaling pathway (Table 4). In the MAPK pathway, there were 14 differentially expressed genes (P < 0.05): DUSP1, GNA12, RAC1, MRAS, ELK1, ATF4, MAPK3, CDC25B, STMN1, MAX, TGFBR2, MAPK11, CACNB3, FGF8, and RASA2 (Table 5). In the TLR pathway, there were 8 key genes with a significant difference in expression (P < 0.05): RAC1, TRAF3, MAPK3, IFNAR1, TICAM1, IL6, MAPK11, and IFNA1 (Table 5). However, there were only two differentially expressed genes (CACNA2D3 and PPM1B) in the MAPK pathway and one key gene (TLR6) in the TLR pathway in the AS group cultured in normal media, both with an insignificant KEGG analysis score (P > 0.05). The results of qRT-PCR verified the expression variation of such genes mentioned above, including GNA12, DUSP1, MAPK11, RAC1, MAPK3, TRAF3, IFNAR1, IL6, and IFNA1 (Figure 3).


Whole Genome Expression Profiling and Signal Pathway Screening of MSCs in Ankylosing Spondylitis.

Li Y, Wang P, Xie Z, Huang L, Yang R, Gao L, Tang Y, Zhang X, Ye J, Chen K, Cai Z, Wu Y, Shen H - Stem Cells Int (2014)

Verification of differentially expressed genes of MSCs in AS patients cultured in inflammatory environment. RNA was isolated from MSCs of HD (n = 5) and AS patients (n = 5) after MSCs were cultured in inflammatory environment mimicked by TNF-α (10 ng/mL) and IFN-γ (10 ng/mL). Comparison of the levels of differentially expressed genes in MSCs from both groups was based on quantitative real-time polymerase chain reaction (qRT-PCR). Beta-actin was used as an internal control. The results showed that when grown in inflammatory culture medium, 9 of 22 differentially expressed genes from whole genomic expression analysis in AS patients were truly differentially expressed. These genes include GNA12, DUSP1, MAPK11, RAC1, MAPK3, TRAF3, IFNAR1, IL6, and IFNA1. The symbol “∗” represents P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4269092&req=5

fig3: Verification of differentially expressed genes of MSCs in AS patients cultured in inflammatory environment. RNA was isolated from MSCs of HD (n = 5) and AS patients (n = 5) after MSCs were cultured in inflammatory environment mimicked by TNF-α (10 ng/mL) and IFN-γ (10 ng/mL). Comparison of the levels of differentially expressed genes in MSCs from both groups was based on quantitative real-time polymerase chain reaction (qRT-PCR). Beta-actin was used as an internal control. The results showed that when grown in inflammatory culture medium, 9 of 22 differentially expressed genes from whole genomic expression analysis in AS patients were truly differentially expressed. These genes include GNA12, DUSP1, MAPK11, RAC1, MAPK3, TRAF3, IFNAR1, IL6, and IFNA1. The symbol “∗” represents P < 0.05.
Mentions: KEGG pathway analysis was used to discover key signaling pathways and relationships between differentially expressed genes. In the inflammatory environment, the two highest scores of indicators in the AS group were hsa04010: mitogen-activated protein kinase (MAPK) signaling pathway, and hsa04620: toll-like receptor (TLR) signaling pathway (Table 4). In the MAPK pathway, there were 14 differentially expressed genes (P < 0.05): DUSP1, GNA12, RAC1, MRAS, ELK1, ATF4, MAPK3, CDC25B, STMN1, MAX, TGFBR2, MAPK11, CACNB3, FGF8, and RASA2 (Table 5). In the TLR pathway, there were 8 key genes with a significant difference in expression (P < 0.05): RAC1, TRAF3, MAPK3, IFNAR1, TICAM1, IL6, MAPK11, and IFNA1 (Table 5). However, there were only two differentially expressed genes (CACNA2D3 and PPM1B) in the MAPK pathway and one key gene (TLR6) in the TLR pathway in the AS group cultured in normal media, both with an insignificant KEGG analysis score (P > 0.05). The results of qRT-PCR verified the expression variation of such genes mentioned above, including GNA12, DUSP1, MAPK11, RAC1, MAPK3, TRAF3, IFNAR1, IL6, and IFNA1 (Figure 3).

Bottom Line: Our results showed that in normal media 676 genes were differentially expressed in AS, 354 upregulated and 322 downregulated, while in an inflammatory environment 1767 genes were differentially expressed in AS, 1230 upregulated and 537 downregulated.In addition, by KEGG pathway analysis, 14 key genes from the MAPK signaling and 8 key genes from the TLR signaling pathway were identified as differentially regulated.The results of qRT-PCR verified the expression variation of the 9 genes mentioned above.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, No. 107 Yanjiang Road West, Guangzhou 510120, China.

ABSTRACT
The pathogenesis of dysfunctional immunoregulation of mesenchymal stem cells (MSCs) in ankylosing spondylitis (AS) is thought to be a complex process that involves multiple genetic alterations. In this study, MSCs derived from both healthy donors and AS patients were cultured in normal media or media mimicking an inflammatory environment. Whole genome expression profiling analysis of 33,351 genes was performed and differentially expressed genes related to AS were analyzed by GO term analysis and KEGG pathway analysis. Our results showed that in normal media 676 genes were differentially expressed in AS, 354 upregulated and 322 downregulated, while in an inflammatory environment 1767 genes were differentially expressed in AS, 1230 upregulated and 537 downregulated. GO analysis showed that these genes were mainly related to cellular processes, physiological processes, biological regulation, regulation of biological processes, and binding. In addition, by KEGG pathway analysis, 14 key genes from the MAPK signaling and 8 key genes from the TLR signaling pathway were identified as differentially regulated. The results of qRT-PCR verified the expression variation of the 9 genes mentioned above. Our study found that in an inflammatory environment ankylosing spondylitis pathogenesis may be related to activation of the MAPK and TLR signaling pathways.

No MeSH data available.


Related in: MedlinePlus