PMID- 29125839 OWN - NLM STAT- MEDLINE DCOM- 20171211 LR - 20181113 IS - 1935-2735 (Electronic) IS - 1935-2727 (Print) IS - 1935-2727 (Linking) VI - 11 IP - 11 DP - 2017 Nov TI - Csseverin inhibits apoptosis through mitochondria-mediated pathways triggered by Ca2 + dyshomeostasis in hepatocarcinoma PLC cells. PG - e0006074 LID - 10.1371/journal.pntd.0006074 [doi] LID - e0006074 AB - 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. Csseverin, a component of the excretory/secretory products of C. sinensis (CsESPs), 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 Csseverin gene. METHODS: In the present study, we evaluated the effects of Csseverin 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 Csseverin 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 Csseverin in PLC cells showed apoptosis resistance after the induction of apoptosis. Additionally, the activation of caspase-3 and caspase-9 was specifically weakened in Csseverin overexpression PLC cells. The overexpression of Csseverin 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 Csseverin overexpression PLC cells. CONCLUSIONS: The present findings suggest that Csseverin, a component of CsESPs, confers protection from human HCC cell apoptosis via the inactivation of membranous Ca2+ channels. Csseverin might be involved in the process of HCC through C. sinensis infestation in affected patients. FAU - Shi, Mengchen AU - Shi M AUID- ORCID: 0000-0002-9457-8211 AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Key Laboratory of Liver Disease Research, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China. FAU - Zhou, Lina AU - Zhou L AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Zhao, Lu AU - Zhao L AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Shang, Mei AU - Shang M AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - He, Tongtong AU - He T AD - School of Public Health, Sun Yat-Sen University, Guangzhou, China. FAU - Tang, Zeli AU - Tang Z AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Sun, Hengchang AU - Sun H AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Ren, Pengli AU - Ren P AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Lin, Zhipeng AU - Lin Z AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Chen, Tingjin AU - Chen T AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Yu, Jinyun AU - Yu J AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Xu, Jin AU - Xu J AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Yu, Xinbing AU - Yu X AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. FAU - Huang, Yan AU - Huang Y AD - Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China. AD - Key Laboratory for Tropical Disease Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, China. AD - Guangdong Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, China. LA - eng PT - Journal Article DEP - 20171110 PL - United States TA - PLoS Negl Trop Dis JT - PLoS neglected tropical diseases JID - 101291488 RN - 0 (Helminth Proteins) SB - IM MH - Animals MH - Apoptosis/*drug effects MH - Cell Line, Tumor MH - Cell Survival/drug effects MH - Clonorchis sinensis/*pathogenicity MH - Helminth Proteins/*metabolism MH - Humans MH - Mitochondria/*drug effects PMC - PMC5705155 COIS- The authors have declared that no competing interests exist. EDAT- 2017/11/11 06:00 MHDA- 2017/12/12 06:00 PMCR- 2017/11/10 CRDT- 2017/11/11 06:00 PHST- 2017/08/01 00:00 [received] PHST- 2017/10/26 00:00 [accepted] PHST- 2017/11/28 00:00 [revised] PHST- 2017/11/11 06:00 [pubmed] PHST- 2017/12/12 06:00 [medline] PHST- 2017/11/11 06:00 [entrez] PHST- 2017/11/10 00:00 [pmc-release] AID - PNTD-D-17-01224 [pii] AID - 10.1371/journal.pntd.0006074 [doi] PST - epublish SO - PLoS Negl Trop Dis. 2017 Nov 10;11(11):e0006074. doi: 10.1371/journal.pntd.0006074. eCollection 2017 Nov.