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Trps1 regulates biliary epithelial-mesenchymal transition and has roles during biliary fibrosis in liver grafts: a preliminary study.

Zhe C, Yu F, Tian J, Zheng S - PLoS ONE (2015)

Bottom Line: Mesenchymal markers were seen in biliary epithelial cells (BECs), with collagen deposited around the bile duct.Expression of epithelial marker mRNAs and proteins in HIBECs decreased with prolonged cold preservation (CP), while mesenchymal marker expression increased.Expression of E-cadherin was increased in HIBECs following Trps1 adenovirus infection and CP/reperfusion injury (CPRI), with vimentin expression levels reduced and CPRI-mediated epithelial-mesenchymal transition (EMT) inhibited.

View Article: PubMed Central - PubMed

Affiliation: Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, No. 29 Gaotanyan Road, Shapingba District, Chongqing, 400038, China.

ABSTRACT

Objective: To investigate the role(s) of Trps1 in non-anastomotic biliary stricture (NABS) following liver transplantation.

Methods: Immunohistochemical and histological techniques were used to detect Trps1, E-cadherin, CK19, vimentin, α-SMA, and collagen deposition. Human intrahepatic biliary epithelial cells (HIBECs) were infected with a Trps1 adenovirus, or transfected with Trps1 short-interfering RNAs (siRNAs). Reverse transcription polymerase chain reaction (RT-PCR) assays and western blotting were used to determine expression levels of epithelial and mesenchymal markers, and Trps1 in HIBECs.

Results: Expression of Trps1 and epithelial markers was down-regulated or absent in NABS liver samples. Mesenchymal markers were seen in biliary epithelial cells (BECs), with collagen deposited around the bile duct. Trps1 expression positively correlated with epithelial markers. Expression of epithelial marker mRNAs and proteins in HIBECs decreased with prolonged cold preservation (CP), while mesenchymal marker expression increased. A 12-h CP period led to increased Trps1 mRNA and protein levels. Expression of E-cadherin was increased in HIBECs following Trps1 adenovirus infection and CP/reperfusion injury (CPRI), with vimentin expression levels reduced and CPRI-mediated epithelial-mesenchymal transition (EMT) inhibited. Transfection of HIBECs with Trps1 siRNAs in conjunction with CPRI revealed that E-cadherin expression was decreased, vimentin expression was increased, and CPRI-mediated EMT was promoted.

Conclusion: Trps1 is involved in NABS pathogenesis following liver transplantation and negatively correlates with BEC EMT and biliary fibrosis in liver grafts. Trps1 demonstrates antagonistic effects that could reverse EMT.

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Related in: MedlinePlus

Western blotting analysis of Trps1 expression changes during CPRI-mediated EMT in HIBECs.A: Western blotting results; B–D: semi-quantitative Trps1, E-cadherin, and Vimentin protein results, respectively. Lane 1: HIBECs; 2: HIBECs subjected to CPRI; 3: HIBECs infected with a Trps1 adenoviurs and subjected to CPRI; 4: HIBECs transfected with a Trps1-specific siRNA, and subjected to CPRI.
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pone.0123233.g007: Western blotting analysis of Trps1 expression changes during CPRI-mediated EMT in HIBECs.A: Western blotting results; B–D: semi-quantitative Trps1, E-cadherin, and Vimentin protein results, respectively. Lane 1: HIBECs; 2: HIBECs subjected to CPRI; 3: HIBECs infected with a Trps1 adenoviurs and subjected to CPRI; 4: HIBECs transfected with a Trps1-specific siRNA, and subjected to CPRI.

Mentions: Cells infected with the Trps1 adenovirus showed increased Trps1 mRNA levels compared with those in uninfected cells or cells infected with the empty control virus. When the infected HIBECs were subjected to CPRI, we observed alterations in the expression levels of genes associated with EMT. There was a significant increase in E-cadherin mRNA and protein levels (p = 0.002 and p < 0.001, respectively), and a significant decrease in Vimentin mRNA and protein levels (p < 0.001, p = 0.017), with CPRI-mediated EMT inhibited.Trps1 mRNA levels were decreased in cells transfected with the Trps1-specific siRNA compared with those in untransfected cells or those transfected with the control siRNA (Fig 5). In contrast, HIBECs transfected with the Trps1-specific siRNA and subjected to CPRI showed significantly decreased mRNA and protein expression levels for E-cadherin (p = 0.032 and 0.044, respectively). Vimentin mRNA and protein levels were significantly increased compared with those in untreated control cells (p = 0.047 and 0.031, respectively), with CPRI-mediated EMT enhanced (Figs 6 and 7).


Trps1 regulates biliary epithelial-mesenchymal transition and has roles during biliary fibrosis in liver grafts: a preliminary study.

Zhe C, Yu F, Tian J, Zheng S - PLoS ONE (2015)

Western blotting analysis of Trps1 expression changes during CPRI-mediated EMT in HIBECs.A: Western blotting results; B–D: semi-quantitative Trps1, E-cadherin, and Vimentin protein results, respectively. Lane 1: HIBECs; 2: HIBECs subjected to CPRI; 3: HIBECs infected with a Trps1 adenoviurs and subjected to CPRI; 4: HIBECs transfected with a Trps1-specific siRNA, and subjected to CPRI.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4401436&req=5

pone.0123233.g007: Western blotting analysis of Trps1 expression changes during CPRI-mediated EMT in HIBECs.A: Western blotting results; B–D: semi-quantitative Trps1, E-cadherin, and Vimentin protein results, respectively. Lane 1: HIBECs; 2: HIBECs subjected to CPRI; 3: HIBECs infected with a Trps1 adenoviurs and subjected to CPRI; 4: HIBECs transfected with a Trps1-specific siRNA, and subjected to CPRI.
Mentions: Cells infected with the Trps1 adenovirus showed increased Trps1 mRNA levels compared with those in uninfected cells or cells infected with the empty control virus. When the infected HIBECs were subjected to CPRI, we observed alterations in the expression levels of genes associated with EMT. There was a significant increase in E-cadherin mRNA and protein levels (p = 0.002 and p < 0.001, respectively), and a significant decrease in Vimentin mRNA and protein levels (p < 0.001, p = 0.017), with CPRI-mediated EMT inhibited.Trps1 mRNA levels were decreased in cells transfected with the Trps1-specific siRNA compared with those in untransfected cells or those transfected with the control siRNA (Fig 5). In contrast, HIBECs transfected with the Trps1-specific siRNA and subjected to CPRI showed significantly decreased mRNA and protein expression levels for E-cadherin (p = 0.032 and 0.044, respectively). Vimentin mRNA and protein levels were significantly increased compared with those in untreated control cells (p = 0.047 and 0.031, respectively), with CPRI-mediated EMT enhanced (Figs 6 and 7).

Bottom Line: Mesenchymal markers were seen in biliary epithelial cells (BECs), with collagen deposited around the bile duct.Expression of epithelial marker mRNAs and proteins in HIBECs decreased with prolonged cold preservation (CP), while mesenchymal marker expression increased.Expression of E-cadherin was increased in HIBECs following Trps1 adenovirus infection and CP/reperfusion injury (CPRI), with vimentin expression levels reduced and CPRI-mediated epithelial-mesenchymal transition (EMT) inhibited.

View Article: PubMed Central - PubMed

Affiliation: Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, No. 29 Gaotanyan Road, Shapingba District, Chongqing, 400038, China.

ABSTRACT

Objective: To investigate the role(s) of Trps1 in non-anastomotic biliary stricture (NABS) following liver transplantation.

Methods: Immunohistochemical and histological techniques were used to detect Trps1, E-cadherin, CK19, vimentin, α-SMA, and collagen deposition. Human intrahepatic biliary epithelial cells (HIBECs) were infected with a Trps1 adenovirus, or transfected with Trps1 short-interfering RNAs (siRNAs). Reverse transcription polymerase chain reaction (RT-PCR) assays and western blotting were used to determine expression levels of epithelial and mesenchymal markers, and Trps1 in HIBECs.

Results: Expression of Trps1 and epithelial markers was down-regulated or absent in NABS liver samples. Mesenchymal markers were seen in biliary epithelial cells (BECs), with collagen deposited around the bile duct. Trps1 expression positively correlated with epithelial markers. Expression of epithelial marker mRNAs and proteins in HIBECs decreased with prolonged cold preservation (CP), while mesenchymal marker expression increased. A 12-h CP period led to increased Trps1 mRNA and protein levels. Expression of E-cadherin was increased in HIBECs following Trps1 adenovirus infection and CP/reperfusion injury (CPRI), with vimentin expression levels reduced and CPRI-mediated epithelial-mesenchymal transition (EMT) inhibited. Transfection of HIBECs with Trps1 siRNAs in conjunction with CPRI revealed that E-cadherin expression was decreased, vimentin expression was increased, and CPRI-mediated EMT was promoted.

Conclusion: Trps1 is involved in NABS pathogenesis following liver transplantation and negatively correlates with BEC EMT and biliary fibrosis in liver grafts. Trps1 demonstrates antagonistic effects that could reverse EMT.

Show MeSH
Related in: MedlinePlus