Identifying Biomarkers to Predict the Prognosis of Biliary Atresia by Weighted Gene Co-Expression Network Analysis

Abstract

The prognosis of children with biliary atresia (BA) after Kasai operation remains difficult to predict, and liver fibrosis is closely related to the prognosis of children with BA. We aimed to find biomarkers for native liver survival (NLS) prediction by weighted gene co-expression network analysis (WGCNA). The biological processes and signal pathways that biomarkers involved in were further analyzed by bioinformatics. Quantitative Real-time PCR, Western blot and immunohistochemistry was performed to detect biomarkers expression. The relationship of biomarkers with clinicopathological characteristics of BA was also investigated. LECT2 was overexpressed or knockdown in LX-2 cells, and the expression of fibrogenic genes such as a-SMA and COL1A1 was quantified. We found that LECT2 mRNA expression was higher in BA liver tissues compared with normal liver tissues. Bioinformatics showed that LECT2 mainly played a fibrosis-promoting role in the development in BA by regulating bile acid metabolism and promoting inflammatory response. LECT2 immunohistochemistry scores of BA children were higher than control group (p = 0.001). Survival analysis revealed that LECT2 high expression is an unfavorable prognostic factor for native liver survival in BA patients. Additionally, the high LECT2 expression was an independent prognostic factor affecting native liver survival (HR 3.702, 95%CI:2.085–6.575, p = 0.001). LECT2 modulates TGF-β mediated a-SMA and COL1A1 expression in LX-2 cells. siRNA-LECT2 inhibits the expression of a-SMA and COL1A1 in LX-2 cells. Overexpression of LECT2 resulted in an increase in a-SMA and COL1A1 expression. Knockdown of LECT2 inhibits the proliferation and increase apoptosis in activated LX-2 cells. LECT2 may act as a new prognostic biomarker for native liver survival in BA patients.

TGF-β1-Mediated Leukocyte Cell-Derived Chemotaxin 2 Is Associated With Liver Fibrosis in Biliary Atresia

“…We believe that it may be related to the mechanism of BA fibrosis. Moreover, it was proposed to be related to liver fibrosis in BA and involved in BA development (5). Thus, we initially evaluated the expression of LECT2 in liver tissue and serum of patients with BA.…”

“…We believe that it may be related to the mechanism of BA fibrosis. Moreover, it was proposed to be related to liver fibrosis in BA and involved in BA development ( 5 ).…”

“…On the other hand, macrophages produce pro-fibrotic mediators such as TGF-β1 and platelet-derived growth factor to regulate the hepatic stellate cells trans-differentiation to the key extracellular matrix producing cells, myofibroblasts ( 25 ). And LECT2 participated in the inflammatory response to promote liver fibrosis and was regulated by TGF-β1 ( 5 ). Therefore, we hypothesize that TGF-β1 is the link between LECT2 and CD163 + macrophage interactions.…”

“…LECT2 plays a crucial role in liver fibrogenesis ( 4 ). It promotes liver fibrosis in BA development by regulating TGF-β-mediated α-SMA and COLIA1 expression in LX-2 cells ( 5 ).…”

Progress in Biomarkers Related to Biliary Atresia

“…Kong et al . 101 found increased expression of leukocyte cell-derived chemotaxin 2 (LECT2) mRNA in the liver tissues of children with BA. Another study found that LECT2 was also highly expressed in the sera of children with BA.…”