MicroRNA-200c regulates cisplatin resistance by targeting ZEB2 in human gastric cancer cells

Jiang,Tao; Dong,Pengfei; Li,Long; Ma,Xiao; Xu,Pei; Zhu,He; Wang,Yanqiu; Yang,Baotong; Liu,Kuangge; Liu,Jinwei; Xue,Juan; Lv,Runzhe; Su,Panke; Kong,Guoqiang; Chang,Yongchao; Zhao,Chonggao; Wang,Lidong

doi:10.3892/or.2017.5659

MicroRNA-200c regulates cisplatin resistance by targeting ZEB2 in human gastric cancer cells

Authors:
- Tao Jiang
- Pengfei Dong
- Long Li
- Xiao Ma
- Pei Xu
- He Zhu
- Yanqiu Wang
- Baotong Yang
- Kuangge Liu
- Jinwei Liu
- Juan Xue
- Runzhe Lv
- Panke Su
- Guoqiang Kong
- Yongchao Chang
- Chonggao Zhao
- Lidong Wang
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Affiliations: Henan Key Laboratory - Esophageal Cancer Laboratory for Cancer Research, Basic Medical College, Zhengzhou University, Zhengzhou, Henan 450052, P.R. China, Department of Clinical Laboratory, The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471013, P.R. China
Published online on: May 22, 2017 https://doi.org/10.3892/or.2017.5659
Pages: 151-158

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Abstract

This study was specifically designed to confirm the hypothesis that microRNA-200c (miR-200c) affects the development of cisplatin (DDP) resistance in human gastric cancer cells by targeting zinc finger E-box binding homeobox 2 (ZEB2). A total of 50 gastric cancer tissues and their corresponding normal adjacent tissue samples were collected. Then, the expression levels of miR-200c and ZEB2 in both gastric cancer specimens and cells were detected using the quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemical methods. A dual‑luciferase reporter gene assay was conducted to evaluate the effect of miR-200c on the 3'-untranslated region (3'UTR) luciferase activity of ZEB2. SGC7901/DDP cells were transfected with miR-200c mimics and ZEB2 siRNA, respectively. Subsequently, changes in cellular proliferation and apoptosis were detected through the methyl thiazolyl tetrazolium assay and flow cytometric analysis, respectively. We also carried out a western blot analysis assay in order to detect the expression of apoptosis-related genes and ZEB2. miR-200c was significantly downregulated and ZEB2 was significantly upregulated in both gastric cancer tissues and SGC7901/DDP cells when compared with those in normal tissues and SGC7901 cells (P<0.01). The dual luciferase reporter gene assay showed that miR-200c could specifically bind with the 3'UTR of ZEB2 and significantly suppress the luciferase activity by 42% (P<0.01). Upregulation of miR-200c or downregulation of ZEB2 enhanced the sensitivity of SGC7901/DDP cells to DDP. miR‑200c was significantly downregulated in both gastric cancer tissues and cells, while the expression of ZEB2 exhibited the opposite trend. Our study further demonstrated that miR-200c could enhance the sensitivity of SGC7901/DDP cells to DDP through targeted regulation of ZEB2 expression in gastric cancer tissues.

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