Tumor Biology

, Volume 37, Issue 11, pp 15315–15324 | Cite as

miR-222 induces Adriamycin resistance in breast cancer through PTEN/Akt/p27kip1 pathway

  • Dan-dan Wang
  • Su-jin Yang
  • Xiu Chen
  • Hong-Yu Shen
  • Long-ji Luo
  • Xiao-hui Zhang
  • Shan-liang Zhong
  • Jian-hua ZhaoEmail author
  • Jin-hai TangEmail author
Original Article


The high resistant rate of Adriamycin (Adr) is associated with a poor prognosis of breast cancer in women worldwide. Since miR-222 might contribute to chemoresistance in many cancer types, in this study, we aimed to investigate its efficacy in breast cancer through PTEN/Akt/p27 kip1 pathway. Firstly, in vivo, we verified that miR-222 was upregulated in chemoresistant tissues after surgery compared with the paired preneoadjuvant samples of 21 breast cancer patients. Then, human breast cancer Adr-resistant cell line (MCF-7/Adr) was constructed to validate the pathway from the parental sensitive cell line (MCF-7/S). MCF-7/Adr and MCF-7/S were transfected with miR-222 mimics, miR-222 inhibitors, or their negative controls, respectively. The results showed that inhibition of miR-222 in MCF-7/Adr significantly increased the expressions of PTEN and p27 kip1 and decreased phospho-Akt (p-Akt) both in mRNA and protein levels (p < 0.05) by using quantitative real-time PCR (qRT-PCR) and western blot. MTT and flow cytometry suggested that lower expressed miR-222 enhanced apoptosis and decreased the IC50 of MCF-7/Adr cells. Additionally, immunofluorescence demonstrated that the subcellular location of p27 kip1 was dislocated resulting from the alteration of miR-222. Conversely, in MCF-7/S transfected with miR-222 mimics, upregulation of miR-222 is associated with decreasing PTEN and p27 kip1 and increasing Akt accompanied by less apoptosis and higher IC50. Importantly, Adr resistance induced by miR-222 overexpression through PTEN/Akt/p27 was completely blocked by LY294002, an Akt inhibitor. Taken together, these data firstly elucidated that miR-222 could reduce the sensitivity of breast cancer cells to Adr through PTEN/Akt/p27 kip1 signaling pathway, which provided a potential target to increase the sensitivity to Adr in breast cancer treatment and further improved the prognosis of breast cancer patients.


miR-222 PTEN/Akt/p27 pathway Adr resistance Breast cancer 



We thank Shan-liang Zhong and Wei-xian Chen for useful discussions and help in revision of the present paper. This study was funded by the National Natural Science Foundation of China (grant number 81272470).

Compliance with ethical standards

Conflicts of interest

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.


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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Dan-dan Wang
    • 1
  • Su-jin Yang
    • 1
  • Xiu Chen
    • 1
  • Hong-Yu Shen
    • 1
  • Long-ji Luo
    • 1
  • Xiao-hui Zhang
    • 2
  • Shan-liang Zhong
    • 2
  • Jian-hua Zhao
    • 2
    Email author
  • Jin-hai Tang
    • 1
    Email author
  1. 1.Department of General SurgeryJiangsu Cancer Hospital Affiliated to Nanjing Medical UniversityNanjingChina
  2. 2.Center of Clinical Laboratory ScienceJiangsu Cancer Hospital Affiliated to Nanjing Medical UniversityNanjingChina

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