Open AccessMolecular Characterization of Severin from Clonorchis sinensis Excretory/Secretory Products and Its Potential Anti-apoptotic Role in Hepatocarcinoma PLC Cells
Author(s) -
Xueqing Chen,
Shan Li,
Lei He,
Xiaoyun Wang,
Pei Liang,
Wenjun Chen,
Meng Bian,
Mengyu Ren,
Jintian Lin,
Chi Liang,
Jin Xu,
Zhongdao Wu,
Xuerong Li,
Yan Huang,
Xinbing Yu
Publication year - 2013
Publication title -
plos neglected tropical diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.99
H-Index - 135
eISSN - 1935-2735
pISSN - 1935-2727
DOI - 10.1371/journal.pntd.0002606
Subject(s) - clonorchis sinensis , biology , apoptosis , microbiology and biotechnology , clonorchiasis , annexin , gelsolin , immunology , biochemistry , actin , helminths
Background Clonorchiasis, caused by the infection of Clonorchis sinensis ( C. sinensis ), is a kind of neglected tropical disease, but it is highly related to cholangiocarcinoma and hepatocellular carcinoma (HCC). It has been well known that the excretory/secretory products of C. sinensis ( Cs ESPs) play key roles in clonorchiasis associated carcinoma. From genome and transcriptome of C. sinensis , we identified one component of Cs ESPs, severin ( Cs severin), which had three putative gelsolin domains. Its homologues are supposed to play a vital role in apoptosis resistance of tumour cell. Methodology/Principal Findings There was significant similarity in tertiary structures between human gelsolin and Cs severin by bioinformatics analysis. We identified that Cs severin expressed at life stage of adult worm, metacercaria and egg by the method of quantitative real-time PCR and western blotting. Cs severin distributed in vitellarium and intrauterine eggs of adult worm and tegument of metacercaria by immunofluorence assay. We obtained recombinant Cs severin (r Cs severin) and confirmed that r Cs severin could bind with calciumion in circular dichroism spectrum analysis. It was demonstrated that r Cs severin was of the capability of actin binding by gel overlay assay and immunocytochemistry. Both Annexin V/PI assay and mitochondrial membrane potential assay of human hepatocarcinoma cell line PLC showed apoptosis resistance after incubation with different concentrations of r Cs severin. Morphological analysis, apoptosis-associated changes of mitochondrial membrane potential and Annexin V/PI apoptosis assay showed that co-incubation of PLC cells with r Cs severin in vitro led to an inhibition of apoptosis induced by serum-starved for 24 h. Conclusions/Significance Collectively, the molecular properties of Cs severin, a molecule of Cs ESPs, were characterized in our study. r Cs severin could cause obvious apoptotic inhibition in human HCC cell line. Cs severin might exacerbate the process of HCC patients combined with C. sinensis infection.