Establishment and characterization of a GES‑1 human gastric epithelial cell line stably expressing miR‑23a

Chen,Li; Gao,Yan; Zhu,Lihua; Song,Hongjiang; Zhao,Linlin; Liu,Aihua; Zhang,Guangling; Shi,Guoyou

doi:10.3892/ol.2018.8765

Establishment and characterization of a GES‑1 human gastric epithelial cell line stably expressing miR‑23a

Authors:
- Li Chen
- Yan Gao
- Lihua Zhu
- Hongjiang Song
- Linlin Zhao
- Aihua Liu
- Guangling Zhang
- Guoyou Shi
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Affiliations: Department of Pathogen Biology, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan, Hebei 063210, P.R. China, The First Department of General Surgery, Hongqi Hospital Affiliated to Mudanjiang Medical University, Mudanjiang, Heilongjiang 157011, P.R. China, Department of Gastrointestinal Surgery, Harbin Medical University Cancer Hospital, Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China, Pharmacy Disciplines, Jitang College, North China University of Science and Technology, Tangshan, Hebei 063210, P.R. China, Department of Pathogen Biology, Jitang College, North China University of Science and Technology, Tangshan, Hebei 063210, P.R. China
Published online on: May 22, 2018 https://doi.org/10.3892/ol.2018.8765
Pages: 977-983
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

MicroRNAs (miRNAs/miRs) are highly conserved, endogenous, small and single‑stranded RNA molecules that promote the degradation and translational inhibition of specific target mRNAs in order to regulate cell proliferation and differentiation, and organism growth and development. MiR‑23a has been demonstrated to function as an oncogene in certain types of tumor. The aim of the present study was to provide a tool for elucidating the mechanisms of action of miR‑23a in gastric cancer, and identify the function of miR‑23a in a human gastric epithelium cell line, by establishing a human gastric epithelial GES‑1 cell line that stably expressed miR‑23a. A plasmid was constructed for the expression of miR‑23a by inserting the miR‑23a primary sequence into a pcDNA3 vector (pcDNA3/pri‑23a). PcDNA3/pri‑23a or the empty pcDNA3 vector (EV), which was then transfected into human gastric epithelium GES‑1 cells using Lipofectamine to produce GES‑1/miR‑23a cells and GES‑1/EV cells, respectively. G418 (Geneticin) was used to select and expand the G418‑resistant colonies, and miR‑23a expression was assessed by reverse transcription‑semi‑quantitative polymerase chain reaction. The proliferation of the cells was assessed using cell counting and MTT assays. The invasive ability of the cells was evaluated using a Transwell assay. The colony‑forming ability of the cells was assessed using a colony formation assay. A human gastric epithelium GES‑1/miR‑23a cell line with the stable expression of miR‑23a was successfully established. Compared with the control GES‑1 and GES‑1/EV cells, the mRNA expression of the miR‑23a gene in GES‑1/miR‑23a cells was significantly increased (P<0.05). The proliferation rate, invasive ability and colony‑forming ability of the GES‑1/miR‑23a cells were significantly higher compared with those of the control GES‑1/EV cells and the parental GES‑1 cells (P<0.05). Additionally, the results of the present study demonstrated that miR‑23a enhanced the cell proliferation rate, invasive ability and cell colony forming ability of GES‑1 cells. This data provides a solid experimental foundation for further studies on the function of miRNAs in the development and progression of gastric cancer.

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