Production of DUSP1 protein using the baculovirus insect cell expression system and its in vitro effects on cancer cells

Cheng,Peng; Zhu,Shuying; Jun,Li; Huang,Lihua; Hong,Yahui

doi:10.3892/ijmm.2015.2179

Production of DUSP1 protein using the baculovirus insect cell expression system and its in vitro effects on cancer cells

Authors:
- Peng Cheng
- Shuying Zhu
- Li Jun
- Lihua Huang
- Yahui Hong
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Affiliations: College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, Hunan 410128, P.R. China
Published online on: April 9, 2015 https://doi.org/10.3892/ijmm.2015.2179
Pages: 1715-1719

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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.

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