Open AccessCsseverin inhibits apoptosis through mitochondria-mediated pathways triggered by Ca2 + dyshomeostasis in hepatocarcinoma PLC cells
Author(s) -
Mei Shi,
Lina Zhou,
Lu Zhao,
Mei Shang,
Tongtong He,
Zeli Tang,
Hengchang Sun,
Pengli Ren,
Zhipeng Lin,
Tingjin Chen,
Jinyun Yu,
Jin Xu,
Xinbing Yu,
Yan Huang
Publication year - 2017
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.0006074
Subject(s) - apoptosis , microbiology and biotechnology , mitochondrial permeability transition pore , biology , transfection , cytochrome c , mitochondrion , clonorchis sinensis , cell culture , programmed cell death , biochemistry , immunology , genetics , helminths
Background Numerous experimental and epidemiological studies have demonstrated a link between Clonorchis sinensis ( C . sinensis ) infestation and cholangiocarcinoma (CCA) as well as hepatocellular carcinoma (HCC). The underlying molecular mechanism involved in the malignancy of CCA and HCC has not yet been addressed. Cs severin, a component of the excretory/secretory products of C. sinensis ( Cs ESPs), was confirmed to cause obvious apoptotic inhibition in the human HCC cell line PLC. However, the antiapoptotic mechanism is unclear. In the present study, we investigated the cellular features of the antiapoptotic mechanism upon transfection of the Cs severin gene. Methods In the present study, we evaluated the effects of Cs severin gene overexpression on the apoptosis of PLC cells using an Annexin PE/7-AAD assay. Western blotting was applied to quantify the activation of caspase-3 and caspase-9, the mitochondrial translocation of Bax and the release of Cyt c upon Cs severin overexpression in PLC cells. Laser scanning confocal microscopy was used to analyze the changes of intracellular calcium. Fluorescence assay and immunofluorescence assays were performed to observe the changes of the mitochondrial permeability transition pore (MPTP). Results The overexpression of Cs severin in PLC cells showed apoptosis resistance after the induction of apoptosis. Additionally, the activation of caspase-3 and caspase-9 was specifically weakened in Cs severin overexpression PLC cells. The overexpression of Cs severin reduced the increase in intracellular free Ca2+, thereby inhibiting MPTP opening in PLC cells. Moreover, Bax mitochondrial translocation and the subsequent release of Cyt c were downregulated in apoptotic Cs severin overexpression PLC cells. Conclusions The present findings suggest that Cs severin, a component of Cs ESPs, confers protection from human HCC cell apoptosis via the inactivation of membranous Ca 2+ channels. Cs severin might be involved in the process of HCC through C . sinensis infestation in affected patients.