title: Inhibitory effects of the recombinant human serine protease inhibitor Hespintor on the proliferation, migration and invasion of hepatocellular carcinoma cells.
abstract: Hespintor is a new Kazal-type serine proteinase inhibitor (Serpin) screened from the HepG2 hepatoblastoma cell line using the suppression subtractive hybridization (SSH) technique. Seprin is closely associated with the progression and remission of malignant tumors, and has certain significance in the diagnosis and treatment of tumors. Investigations on the antitumor activity of Serpin are expected to aid in the development of a new method for tumor treatment based on the serine protease inhibitor. Although the Hespintor prokaryotic expression strain and recombinant Hespintor protein (recombinant fusion protein of Hespintor and rHespintor) have already been obtained, the protein extraction efficiency is low due to the low initial amount of extracted protein and large number of purification steps, which affect the study of the protein function. The aim of the present study was to improve the purification method of rHespintor, increase the protein extraction efficiency, and investigate its effects on the proliferation, migration and invasion of the HepG2 hepatoblastoma cell line. The results demonstrated that the application of urea gradient washing of inclusion body of the protein may effectively remove the majority of impure proteins from the targeted protein. After one-step purification, the target protein rHespintor exhibited a high inhibitory effect of Trypsin Hydrolysis, which was exhibited in a dose-dependent manner. Hoechst 33258 staining was used to determine cell apoptosis. After treating HepG2 hepatoblastoma cells with rHespintor, the cell growth was inhibited, the proliferation ability was reduced, and the number of migrated and invaded cells were significantly decreased. Hoechst 33258 staining and flow cytometry assay results showed clear cell apoptosis. The results reveal showed that rHespintor significantly inhibited proliferation, migration and invasion of the HepG2 hepatoblastoma cell line in vitro, and induced cell apoptosis to a certain extent, indicating that the recombinant protein Hespintor exerts an antitumor effect in vitro, and has the potential and feasibility to become an antitumor drug.
authors.affiliation: Liaoning Provincial University Key Laboratory of Biophysics, College of Medicine, Dalian University, Dalian, Liaoning 116622, P.R. China.
authors.forename: Xue-Lei
authors.initials: XL
authors.lastname: Wang
authors.affiliation: Beijing Center for Physical and Chemical Analysis, Beijing 100094, P.R. China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Li
authors.affiliation: Beijing Center for Physical and Chemical Analysis, Beijing 100094, P.R. China.
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Cheng
authors.affiliation: Liaoning Provincial University Key Laboratory of Biophysics, College of Medicine, Dalian University, Dalian, Liaoning 116622, P.R. China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Chi
authors.affiliation: Liaoning Provincial University Key Laboratory of Biophysics, College of Medicine, Dalian University, Dalian, Liaoning 116622, P.R. China.
authors.forename: Xiao-Kun
authors.initials: XK
authors.lastname: Yao
authors.affiliation: Beijing Center for Physical and Chemical Analysis, Beijing 100094, P.R. China.
authors.forename: Hui-Juan
authors.initials: HJ
authors.lastname: Wu
authors.affiliation: Liaoning Provincial University Key Laboratory of Biophysics, College of Medicine, Dalian University, Dalian, Liaoning 116622, P.R. China.
authors.forename: Yong-Zhi
authors.initials: YZ
authors.lastname: Lun
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D056945:Hep G2 Cells; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry; D008113:Liver Neoplasms / Q000473:pathology*; D009361:Neoplasm Invasiveness; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology*; D015843:Serpins / Q000302:isolation & purification* / Q000494:pharmacology*
chemical_list: D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D015843:Serpins