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Tumor Biology

, Volume 34, Issue 1, pp 349–357 | Cite as

The expression and function of microRNA-203 in lung cancer

  • Jianhua Jin
  • Jianzhong Deng
  • Fang Wang
  • Xiyi Xia
  • Tiefeng Qiu
  • Wenbin Lu
  • Xianwen Li
  • Hua Zhang
  • Xiaoyan Gu
  • Yungang Liu
  • Weiguo Cao
  • Wenlong Shao
Research Article

Abstract

We aimed to determine the expression of microRNA-203 (miR-203) in human lung cancer cell lines and to evaluate the effects of miR-203 by targeting survivin, on the lung cancer cell line 95-D to provide potential new strategies for treating lung cancer. The expression of miR-203 was detected using quantitative real-time PCR (qRT-PCR) in the in vitro cultured lung cancer cells A549, HCC827, NCI-H1299, and 95-D as well as in normal human bronchial epithelial cells. Following a 72-h transfection with the miR-203 precursor in 95-D lung cancer cells, the change in miR-203 expression was detected using qRT-PCR and the resulting effect on survivin protein expression was ascertained by Western blot analysis. The influence of miR-203 on the viability of 95-D lung cancer cells was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The effect of miR-203 on 95-D cell proliferation was analyzed using flow cytometry. The consequences of miR-203 expression on 95-D cell apoptosis were analyzed by Annexin V/propidium iodide double staining coupled with flow cytometry. The role of miR-203 in the invasive potential of 95-D cells was studied using a transwell chamber assay. A luciferase reporter gene system was used to verify that survivin is a target gene for miR-203. By qRT-PCR, the expression of miR-203 was lower in lung cancer cells than in normal bronchial epithelial cells (p < 0.01), and the expression of miR-203 in 95-D lung cancer cells was significantly higher after a 72-h transfection with the miR-203 precursor (p < 0.01). After a 72-h transfection with the miR-203 precursor, survivin protein levels in 95-D cells were significantly decreased (p < 0.01). Cell viability, as assessed with an MTT assay, decreased following an increase in miR-203 expression (p < 0.05). The flow cytometry results indicated that after miR-203 expression increased, the cell proliferation index decreased (p < 0.05) and the number of apoptotic cells increased (p < 0.01). Increased miR-203 expression led to a significant decrease in the number of cells that migrated through a transwell chamber membrane (p < 0.01). The luciferase reporter gene system demonstrated that the relative luciferase activity significantly decreased after transfection with the miR-203 precursor (p < 0.05). The expression of miR-203 is downregulated in lung cancer cells. miR-203 negatively regulates survivin protein expression and inhibits the proliferation and invasion of lung cancer cells. Therapeutic strategies that enhance miR-203 expression or silence survivin could potentially benefit lung cancer patients.

Keywords

miR-203 Lung cancer Survivin Proliferation Apoptosis 

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2012

Authors and Affiliations

  • Jianhua Jin
    • 1
  • Jianzhong Deng
    • 1
  • Fang Wang
    • 1
  • Xiyi Xia
    • 2
  • Tiefeng Qiu
    • 2
  • Wenbin Lu
    • 1
  • Xianwen Li
    • 1
  • Hua Zhang
    • 1
  • Xiaoyan Gu
    • 1
  • Yungang Liu
    • 1
  • Weiguo Cao
    • 3
  • Wenlong Shao
    • 4
  1. 1.Department of Medical Oncology, Wujin People’s HospitalJiangsu UniversityChangzhouPeople’s Republic of China
  2. 2.Department of Respiratory Medicine, Wujin People’s HospitalJiangsu UniversityChangzhouChina
  3. 3.Department of Oncology, Ruijin HospitalShanghai Jiaotong UniversityShanghaiChina
  4. 4.Department of Cardiothoracic SurgeryThe First Affiliated Hospital of Guangzhou Medical CollegeGuangzhouChina

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