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Tumor Biology

, Volume 37, Issue 7, pp 9493–9501 | Cite as

Silencing A7-nAChR levels increases the sensitivity of gastric cancer cells to ixabepilone treatment

  • Chao-Chiang Tu
  • Chien-Yu Huang
  • Wan-Li Cheng
  • Chin-Sheng Hung
  • Yu-Jia Chang
  • Po-Li Wei
Original Article
  • 244 Downloads

Abstract

Gastric cancer is an important health issue worldwide. Currently, improving the therapeutic efficacy of chemotherapy drugs is an important goal of cancer research. Alpha-7 nicotine acetylcholine receptor (A7-nAChR) is the key molecule that mediates gastric cancer progression, metastasis, and therapy responses; however, the role of A7-nAChR in the therapeutic efficacy of ixabepilone remains unclear. A7-nAChR expression was silenced by small interfering RNA (siRNA) technology. The cytotoxicity of ixabepilone was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and ixabepilone-induced apoptosis was analyzed by flow cytometry and annexin V/propidium iodide (PI) apoptotic assay. The expression patterns of anti-apoptotic proteins (AKT, phospho-AKT, Mcl-1, and Bcl-2) and pro-apoptotic proteins (Bad and Bax) were determined by western blot. Our study found that A7-nAChR knockdown (A7-nAChR-KD) AGS cells were more sensitive to ixabepilone administration than scrambled control AGS cells. We found that A7-nAChR knockdown enhanced ixabepilone-induced cell death as evidenced by the increased number of annexin V-positive (apoptotic) cells. After scrambled control and A7-nAChR-KD cells were treated with ixabepilone, we found that pAKT and AKT levels were significantly reduced in both groups of cells. The levels of Bcl-2 and the anti-apoptotic Mcl-1 isoform increased dramatically after ixabepilone treatment in scrambled control cells but not in A7-nAChR-KD cells. Bad and Bax levels did not change between the treatment group and vehicle group in both A7-nAChR-KD and scrambled control cells, whereas cleaved PARP levels dramatically increased in ixabepilone-treated A7-nAChR-KD cells. Our results demonstrated that knockdown of A7-nAChR enhanced the sensitivity of gastric cancer cells to ixabepilone administration. Thus, the A7-nAChR expression level in patients with gastric cancer may be a good indicator of ixabepilone sensitivity.

Keywords

Nicotine acetylcholine receptor Gastric cancer Ixabepilone 

Abbreviation

A7-nAChR

Alpha-7 nicotine acetylcholine receptor

Notes

Acknowledgments

This work was supported by a grant from the National Science Council, Taiwan (Grant No. NSC101-2314-B-038-016-MY3).

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  1. 1.Graduate Institute of Clinical Medicine, College of MedicineTaipei Medical UniversityTaipeiTaiwan
  2. 2.Division of General Surgery, Department of SurgeryNew Taipei HospitalTaipeiTaiwan
  3. 3.Division of General Surgery, Department of Surgery, Shuang Ho HospitalTaipei Medical UniversityTaipeiTaiwan
  4. 4.Department of Surgery, School of Medicine, College of MedicineTaipei Medical UniversityTaipeiTaiwan
  5. 5.Division of General Surgery, Department of SurgeryTaipei Medical University Hospital, Taipei Medical UniversityTaipeiTaiwan
  6. 6.Cancer Research CenterTaipei Medical University Hospital, Taipei Medical UniversityTaipei CityTaiwan

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