ASPSCR-1 and Sirt-5 alleviate Clonorchis liver fluke rCs NOSIP-induced oxidative stress, proliferation, and migration in cholangiocarcinoma cells.

Autor: Bian, Meng, Li, Shan, Zhou, Hanzong, Bi, Lijun, Shen, Yong, Tingjin, Chen, Yu, Xinbing, Huang, Yan, Xu, Qingxia
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Zdroj: PLoS Neglected Tropical Diseases; 11/10/2023, Vol. 17 Issue 11, p1-14, 14p
Abstrakt: 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. It has been well known that NO from chronic inflammation responses are thought to be a major component of the damage and ultimate carcinogenesis ESPs such as nitric oxide synthase interacting protein (NOSIP) are thought to enhance the damage. The objective of this study was to identify the protein candidates interact with recombinant CsNOSIP (rCsNOSIP) and explore their role involved in CCA development or progression. Methods: We applied HuProt microarray containing 21,000 probe sets for a systematic identification of rCsNOSIP-binding proteins and grouped binding hits by gene function. Pull-down assays were used to confirm the interaction of rCsNOSIP with alveolar soft part sarcoma (ASPSCR-1) and sirtuins 5 (Sirt-5). ASPSCR-1/Sirt-5 over-expression and siRNA knockdown experiments were employed for obtain of ASPSCR-1/Sirt-5 high or low expression (ASP-oe/Sirt5-oe or ASP-si/Sirt5-si) cholangiocarcinoma cell line (CCLP-1) cells. Nitric oxide (NO) and reactive oxygen species assay (ROS) as well as cell proliferation and wound-healing assays were performed to observe the effect of rCsNOSIP on ASP-oe/Sirt5-oe or ASP-si/Sirt5-si CCLP-1 cells. Results: Seventy candidate proteins protein "hits" were detected as rCsNOSIP-binding proteins by HuProt microarray and bioinformatics analysis. Pull down assay showed that ASPSCR-1 and Sirt-5 could interact with rCsNOSIP. In addition, endotoxin-free-rCsNOSIP could increase the production of NO and ROS and promote the migration of CCLP-1 cells, while its effect on enhancing cell proliferation was not significant. Furthermore, ROS/NO production, proliferation, or migration were increased in ASP-si or Sirt5-si CCLP-1 cells but decreased in Asp-oe or Sirt5-oe CCLP-1 cells when stimulated with rCsNOSIP. Conclusions: Our findings suggest that CsNOSIP as a component of CsESPs might promote the development and invasion of CCA and Sirt5/ ASPSCR1 as host molecules might play a novel protective role against adverse stimulus during C. sinensis infection. This work supports the idea that CsESPs induce the occurrence and progression of CCA through ROS/RNS-induced oxidative and nitrative DNA damage. Author summary: Clonorchis sinensis (C. sinensis) is prevalent in China, Korea, and Vietnam, and 15–20 million people are estimated to be infected by this fluke, creating a socio-economic burden in epidemic regions. It is generally believed that excretory-secretory products from C. sinensis (CsESPs) can directly interact with the biliary epithelium and induce host inflammatory response. Our previous studies have manifested that CsNOSIP as a component of CsESPs can trigger nitrative and oxidative stress by enhancing the release of NO and reactive oxygen species (ROS) in macrophage. In the current study, we identified rCsNOSIP-binding proteins by HuProt microarray. In addition, we demonstrated that ASPSCR-1 and Sirt-5 could interact with rCsNOSIP by pull down assay. Moreover, rCsNOSIP could increase the production of NO and ROS and promote the migration of human CCLP-1 cells by using NO assay, detection of ROS assay as well as wound-healing assays. Our study provided new information about rCsNOSIP in promoting CCA invasion and metastasis as well as a novel protective role of SIRT5/ ASPSCR1 against adverse stimulus and support a basis for further discovery of drug targets for prevention and control of clonorchiasis. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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