title: Production of DUSP1 protein using the baculovirus insect cell expression system and its in vitro effects on cancer cells.
abstract: The aim of the present study was to produce the human dual specificity phosphatase 1 (DUSP1) protein with biological activity and to investigate its in vitro effects on cancer cells. DUSP1 protein was expressed in the baculovirus expression system and purified by Ni-affinity chromatography followed by dialysis in PBS. The purified protein was verified by SDS-PAGE and western blot analysis. Six cancer cell lines were then cultured in the presence of DUSP1 for various periods of time, and the phosphorylated extracellular signal-regulated kinase (p-ERK) content in each cell line was subsequently determined by western blot analysis. Compared to the β-actin level, the amount of p-ERK markedly decreased after 1 h, indicating that DUSP1 suppressed the expression of p-ERK in 6 cancer cell lines examined. Human cervical cancer cells were also collected and counted following co-culture with DUSP1 to examine its effect on the growth rate of cancer cells. A baculovirus expression system for the production of DUSP1 protein was successfully constructed. The p-ERK content was found to be significantly decreased when the cancer cell lines were exposed to DUSP1. The capability of binary fission was reduced when the cells were examined under a microscope. The proliferation of human cervical cancer cells was also inhibited by DUSP1.
authors: College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, Hunan 410128, P.R. China.
authors: Peng
authors: P
authors: Cheng
authors: College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, Hunan 410128, P.R. China.
authors: Shuying
authors: S
authors: Zhu
authors: College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, Hunan 410128, P.R. China.
authors: Li
authors: L
authors: Jun
authors: College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, Hunan 410128, P.R. China.
authors: Lihua
authors: L
authors: Huang
authors: College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, Hunan 410128, P.R. China.
authors: Yahui
authors: Y
authors: Hong
mesh_terms: D000818:Animals; D016367:Baculoviridae; D049109:Cell Proliferation / Q000187:drug effects*; D054638:Dual Specificity Phosphatase 1 / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D009369:Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D061987:Sf9 Cells; D018411:Spodoptera
chemical_list: D011994:Recombinant Proteins; C517019:DUSP1 protein, human; D054638:Dual Specificity Phosphatase 1