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Tumor Biology

, Volume 34, Issue 2, pp 963–971 | Cite as

Expression and regulatory function of miRNA-34a in targeting survivin in gastric cancer cells

  • Weiguo Cao
  • Rong Fan
  • Lifu Wang
  • Shidan Cheng
  • Hao Li
  • Jinsong Jiang
  • Mei Geng
  • Yening Jin
  • Yunlin Wu
Research Article

Abstract

We aimed to investigate the expression of microRNA-34a (miR-34a) in human gastric cancer cells and to evaluate the effects of miR-34a, acting via its gene survivin, on gastric cancer cell HGC-27 to provide potential new strategies for treating gastric cancer. In vitro cultures of the human gastric cancer cell lines MGC80-3, HGC-27, NCI-N87, and SGC-7901 and the normal human gastric epithelial cell line GES-1 were established. The expression of miR-34a in each gastric cancer cell line and GES-1 normal human gastric epithelial cell line was detected using quantitative real-time polymerase chain reaction (qRT-PCR). After the HGC-27 cells were transfected with a miR-34a mimic for 48 h, the changes in the expression levels of miR-34a were detected using qRT-PCR. The effect of miR-34a on HGC-27 cell viability was measured using a tetrazolium-based colorimetric [−(4,5)-dimethylthiahiazo-(−z-y1)-3,5-di-phenytetrazoliumromide (MTT)] assay. Flow cytometry was used to analyze the effects of miR-34a on HGC-27 cell proliferation. Annexin V/propidium iodide double staining and flow cytometry were used to analyze the effects of miR-34a on HGC-27 cell apoptosis. A Transwell invasion chamber was used to detect the effects of miR-34a on HGC-27 cell invasion. Finally, western blotting was used to analyze the effects of miR-34a on survivin protein expression. The qRT-PCR test determined that miR-34a expression in gastric cancer cells was significantly reduced compared to the normal gastric epithelial cell line GES-1 (p < 0.01). Compared to the control group, cellular miR-34a expression levels were significantly increased in HGC-27 human gastric carcinoma cells after transfection with a miR-34a mimic for 48 h (p < 0.01). The MTT assay demonstrated that after overexpressing miR-34a in HGC-27 cells, cellular viability was significantly reduced (p < 0.05). Flow cytometry analysis determined that upon miR-34a overexpression, the proliferation index decreased significantly (p < 0.05), and cellular apoptosis was significantly increased (p < 0.01). The Transwell invasion chamber assay illustrated that after increasing the expression of miR-34a, the number of cells passing through the Transwell chamber was significantly reduced (p < 0.01). Based on western blotting, compared with the control group, survivin protein expression levels were significantly decreased in the HGC-27 cells transfected with the miR-34a mimic for 48 h (p < 0.01). In conclusion, the expression level of miR-34a was downregulated in human gastric cancer cell lines. miR-34a can negatively regulate survivin protein expression and inhibit gastric cancer cell proliferation and invasion. Therapeutically enhancing miR-34a expression or silencing the survivin gene may benefit patients with gastric cancer.

Keywords

Gastric cancer Survivin MicroRNA-34a HGC-27 

Notes

Conflicts of interest

The authors have no commercial, proprietary, or financial interest in the products or companies described in this article.

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2012

Authors and Affiliations

  • Weiguo Cao
    • 1
    • 2
  • Rong Fan
    • 2
  • Lifu Wang
    • 2
  • Shidan Cheng
    • 2
  • Hao Li
    • 1
  • Jinsong Jiang
    • 1
  • Mei Geng
    • 1
  • Yening Jin
    • 1
  • Yunlin Wu
    • 2
  1. 1.Department of Oncology, Ruijin HospitalShanghai Jiaotong University School of MedicineShanghaiChina
  2. 2.Department of Gastroenterology, Ruijin HospitalShanghai Jiaotong University School of MedicineShanghaiChina

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