Downregulation of AIF-2 Inhibits Proliferation, Migration, and Invasion of Human Glioma Cells via Mitochondrial Dysfunction

  • Wei ChenEmail author
  • Hao Liu
  • Tuo Wang
  • Gang Bao
  • Ning Wang
  • Rui-Chun Li


Glioma remains the leading cause of brain tumor–related death worldwide. Apoptosis inducing factor (AIF) is a family of mitochondrial oxidoreductases that play important roles in mitochondrial metabolism and redox control. AIF-1 has been demonstrated to exert cell-killing effect via apoptosis in cancer cells, whereas the role of AIF-2 in cancer cells has not been determined. This study aimed to investigate the role of AIF-2 in human glioma cells. We found that AIF-2 was upregulated in human glioma tissues and cell lines, especially in U251 cells. Downregulation of AIF-2 using specific siRNA (Si-AIF-2) significantly reduced cell proliferation, induced G1 cell cycle arrest and differently regulated the expression of cell cycle regulator proteins in U251 cells. In addition, the results of Matrigel invasion assay and live-cell tracking assay showed that knockdown of AIF-2 inhibited cell invasion and migration. The results of immunocytochemistry indicated that knockdown of AIF-2 significantly attenuated the nuclear translocation of AIF-1, which was confirmed by western blot analysis. Furthermore, downregulation of AIF-2 resulted in mitochondrial dysfunction in U251 cells, as evidenced by reduced mitochondrial membrane potential (MMP), mitochondrial complex I activity, and mitochondrial Ca2+ buffering capacity. In conclusion, we found that AIF-2 plays a key role in promoting cell proliferation, invasion, and migration via regulating AIF-1-related mitochondrial cascades. Downregulation of the candidate oncogene AIF-2 might constitute a strategy to kill human glioma cells.


Glioma AIF Mitochondrial dysfunction Apoptosis 



The authors would like to thank Dr. Terry Chen for his technical support for the experiments and the preparation of the manuscript.

Compliance with Ethical Standards

This study was approved by the Human Research Ethics Committee of Xi’an Jiaotong University.

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. Bano D, Prehn JHM (2018) Apoptosis-inducing factor (AIF) in physiology and disease: the tale of a repented natural born killer. EBioMedicine 30:29–37CrossRefGoogle Scholar
  2. Boussiotis VA, Charest A (2018) Immunotherapies for malignant glioma. Oncogene 37:1121–1141CrossRefGoogle Scholar
  3. Churbanova IY, Sevrioukova IF (2008) Redox-dependent changes in molecular properties of mitochondrial apoptosis-inducing factor. J Biol Chem 283:5622–5631CrossRefGoogle Scholar
  4. Cloughesy TF, Cavenee WK, Mischel PS (2014) Glioblastoma: from molecular pathology to targeted treatment. Annu Rev Pathol 9:1–25CrossRefGoogle Scholar
  5. Dai B, Hu Z, Huang H, Zhu G, Xiao Z, Wan W, Zhang P, Jia W, Zhang L (2014) Overexpressed KDM5B is associated with the progression of glioma and promotes glioma cell growth via downregulating p21. Biochem Biophys Res Commun 454:221–227CrossRefGoogle Scholar
  6. Fukami T, Nakasu S, Baba K, Nakajima M, Matsuda M (2004) Hyperthermia induces translocation of apoptosis-inducing factor (AIF) and apoptosis in human glioma cell lines. J Neuro-Oncol 70:319–331CrossRefGoogle Scholar
  7. Hangen E, De Zio D, Bordi M, Zhu C, Dessen P, Caffin F, Lachkar S, Perfettini JL, Lazar V, Benard J, Fimia GM, Piacentini M, Harper F, Pierron G, Vicencio JM, Benit P, de Andrade A, Hoglinger G, Culmsee C, Rustin P, Blomgren K, Cecconi F, Kroemer G, Modjtahedi N (2010) A brain-specific isoform of mitochondrial apoptosis-inducing factor: AIF2. Cell Death Differ 17:1155–1166CrossRefGoogle Scholar
  8. Huang ST, Bi KW, Kuo HM, Lin TK, Liao PL, Wang PW, Chuang JH, Liou CW (2014) Phyllanthus urinaria induces mitochondrial dysfunction in human osteosarcoma 143B cells associated with modulation of mitochondrial fission/fusion proteins. Mitochondrion 17:22–33CrossRefGoogle Scholar
  9. Jeong JC, Shin WY, Kim TH, Kwon CH, Kim JH, Kim YK, Kim KH (2011) Silibinin induces apoptosis via calpain-dependent AIF nuclear translocation in U87MG human glioma cell death. J Exp Clin Cancer Res 30:44CrossRefGoogle Scholar
  10. Lee JW, Jeong EG, Soung YH, Kim SY, Nam SW, Kim SH, Lee JY, Yoo NJ, Lee SH (2006) Immunohistochemical analysis of apoptosis-inducing factor (AIF) expression in gastric carcinomas. Pathol Res Pract 202:497–501CrossRefGoogle Scholar
  11. Loeffler M, Daugas E, Susin SA, Zamzami N, Metivier D, Nieminen AL, Brothers G, Penninger JM, Kroemer G (2001) Dominant cell death induction by extramitochondrially targeted apoptosis-inducing factor. FASEB J 15:758–767CrossRefGoogle Scholar
  12. Lopez J, Tait SW (2015) Mitochondrial apoptosis: killing cancer using the enemy within. Br J Cancer 112:957–962CrossRefGoogle Scholar
  13. Millan A, Huerta S (2009) Apoptosis-inducing factor and colon cancer. J Surg Res 151:163–170CrossRefGoogle Scholar
  14. Molenaar JJ, Ebus ME, Koster J, van Sluis P, van Noesel CJ, Versteeg R, Caron HN (2008) Cyclin D1 and CDK4 activity contribute to the undifferentiated phenotype in neuroblastoma. Cancer Res 68:2599–2609CrossRefGoogle Scholar
  15. Motiani RK, Hyzinski-Garcia MC, Zhang X, Henkel MM, Abdullaev IF, Kuo YH, Matrougui K, Mongin AA, Trebak M (2013) STIM1 and Orai1 mediate CRAC channel activity and are essential for human glioblastoma invasion. Pflugers Arch 465:1249–1260CrossRefGoogle Scholar
  16. Preta G (2017) Understanding the Dr. Jekyll and Mr. Hyde nature of apoptosis-inducing factor: future perspectives. Biom J 40:239–240Google Scholar
  17. Sevrioukova IF (2009) Redox-linked conformational dynamics in apoptosis-inducing factor. J Mol Biol 390:924–938CrossRefGoogle Scholar
  18. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA Cancer J Clin 65:87–108CrossRefGoogle Scholar
  19. Vahsen N, Cande C, Briere JJ, Benit P, Joza N, Larochette N, Mastroberardino PG, Pequignot MO, Casares N, Lazar V, Feraud O, Debili N, Wissing S, Engelhardt S, Madeo F, Piacentini M, Penninger JM, Schagger H, Rustin P, Kroemer G (2004) AIF deficiency compromises oxidative phosphorylation. EMBO J 23:4679–4689CrossRefGoogle Scholar
  20. Xie Y, Wanggou S, Liu Q, Li X, Liu J, Wu M (2018) A brain-specific isoform of apoptosis-inducing factor 2 attenuates ischemia-induced oxidative stress in HT22 cells. Neurochem Int 112:179–186CrossRefGoogle Scholar
  21. Yu CJ, Jia LT, Meng YL, Zhao J, Zhang Y, Qiu XC, Xu YM, Wen WH, Yao LB, Fan DM, Jin BQ, Chen SY, Yang AG (2006) Selective proapoptotic activity of a secreted recombinant antibody/AIF fusion protein in carcinomas overexpressing HER2. Gene Ther 13:313–320CrossRefGoogle Scholar
  22. Yu Z, Liu N, Li Y, Xu J, Wang X (2013) Neuroglobin overexpression inhibits oxygen-glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons. Neurobiol Dis 56:95–103CrossRefGoogle Scholar
  23. Zhao H, Wang C, Lu B, Zhou Z, Jin Y, Wang Z, Zheng L, Liu K, Luo T, Zhu D, Chi G, Luo Y, Ge P (2016) Pristimerin triggers AIF-dependent programmed necrosis in glioma cells via activation of JNK. Cancer Lett 374:136–148CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of NeurosurgeryThe First Affiliated Hospital of Xi’an Jiaotong UniversityXi’anChina

Personalised recommendations