Publication: RNAi screen reveals a role of SPHK2 in dengue virus–mediated apoptosis in hepatic cell lines
Issued Date
2017-11-01
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ISSN
19326203
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2-s2.0-85034237715
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Mahidol University
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SCOPUS
Bibliographic Citation
PLoS ONE. Vol.12, No.11 (2017)
Suggested Citation
Atthapan Morchang, Regina Ching Hua Lee, Pa Thai Yenchitsomanus, Gopinathan Pillai Sreekanth, Sansanee Noisakran, Justin Jang Hann Chu, Thawornchai Limjindaporn RNAi screen reveals a role of SPHK2 in dengue virus–mediated apoptosis in hepatic cell lines. PLoS ONE. Vol.12, No.11 (2017). doi:10.1371/journal.pone.0188121 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/41331
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Title
RNAi screen reveals a role of SPHK2 in dengue virus–mediated apoptosis in hepatic cell lines
Abstract
© 2017 Morchang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Hepatic dysfunction is a feature of dengue virus (DENV) infection. Hepatic biopsy specimens obtained from fatal cases of DENV infection show apoptosis, which relates to the pathogenesis of DENV infection. However, how DENV induced liver injury is not fully understood. In this study, we aim to identify the factors that influence cell death by employing an apoptosis-related siRNA library screening. Our results show the effect of 558 gene silencing on caspase 3-mediated apoptosis in DENV-infected Huh7 cells. The majority of genes that contributed to apoptosis were the apoptosis-related kinase enzymes. Tumor necrosis factor superfamily member 12 (TNFSF12), and sphingosine kinase 2 (SPHK2), were selected as the candidate genes to further validate their influences on DENV-induced apoptosis. Transfection of siRNA targeting SPHK2 but not TNFSF12 genes reduced apoptosis determined by Annexin V/PI staining. Knockdown of SPHK2 did not reduce caspase 8 activity; however, did significantly reduce caspase 9 activity, suggesting its involvement of SPHK2 in the intrinsic pathway of apoptosis. Treatment of ABC294649, an inhibitor of SPHK2, reduced the caspase 3 activity, suggesting the involvement of its kinase activity in apoptosis. Knockdown of SPHK2 significantly reduced caspase 3 activity not only in DENV-infected Huh7 cells but also in DENV-infected HepG2 cells. Our results were consistent across all of the four serotypes of DENV infection, which supports the pro-apoptotic role of SPHK2 in DENV-infected liver cells.