title: Expression of Clonorchis sinensis GIIIsPLA2protein in baculovirus-infected insect cells and its overexpression facilitating epithelial-mesenchymal transition in Huh7 cells via AKT pathway.
abstract: Although prior studies confirmed that group III secretory phospholipase A2of Clonorchis sinensis (CsGIIIsPLA2) had stimulating effect on liver fibrosis by binding to LX-2 cells, large-scale expression of recombinant protein and its function in the progression of hepatoma are worth exploring. Because of high productivity and low lipopolysaccharides (LPS) in the Sf9-baculovirus expression system, we firstly used this system to express the coding region of CsGIIIsPLA2. The molecular weight of recombinant CsGIIIsPLA2protein was about 34 kDa. Further investigation showed that most of the recombinant protein presented intracellular expression in Sf9 insect cell nucleus and could be detected only into cell debris, which made the protein purification and further functional study difficult. Therefore, to study the role of CsGIIIsPLA2in hepatocellular carcinoma (HCC) progression, CsGIIIsPLA2overexpression Huh7 cell model was applied. Cell proliferation, migration, and the expression level of epithelial-mesenchymal transition (EMT)-related molecules (E-cadherin, N-cadherin, α-catenin, Vimentin, p300, Snail, and Slug) along with possible mechanism were measured. The results indicated that CsGIIIsPLA2overexpression not only inhibited cell proliferation and promoted migration and EMT but also enhanced the phosphorylation of AKT in HCC cells. In conclusion, this study supported that CsGIIIsPLA2overexpression suppressed cell proliferation and induced EMT through the AKT pathway.
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Mei
authors.initials: M
authors.lastname: Shang
authors.affiliation: Department of Clinical Laboratory, Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Zhizhi
authors.initials: Z
authors.lastname: Xie
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Zeli
authors.initials: Z
authors.lastname: Tang
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Department of Clinical Laboratory, Guangzhou Red Cross Hospital, Medical College, Jinan University, Guangzhou, 510060, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: He
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Caiqin
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Yinjuan
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Lu
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Zhiyue
authors.initials: Z
authors.lastname: Lv
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Zhongdao
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China.
authors.forename: Xinbing
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China. xuerong2@mail.sysu.edu.cn.|Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, People's Republic of China. xuerong2@mail.sysu.edu.cn.
authors.forename: Xuerong
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D016367:Baculoviridae; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation / Q000187:drug effects; D003004:Clonorchis sinensis / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition / Q000502:physiology*; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007313:Insecta; D011485:Protein Binding; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D015801:Helminth Proteins; D014157:Transcription Factors
keywords: Clonorchis sinensis; Epithelial-mesenchymal transition (EMT); Proliferation; Secretory phospholipase A2; Sf9 expression system