Atg7 deficiency increases resistance of MCF-7 human breast cancer cells to photodynamic therapy.
Photodynamic therapy (PDT) uses a photosensitizer, light and oxygen to produce extensive oxidative damage to organelles housing the photosensitizer. Although PDT is an efficient trigger of apoptosis, it also induces autophagy in many kinds of cells. Autophagy can serve as both a cell survival and a cell death mechanism. Our previous study indicates that autophagy contributes to cell death after PDT, especially in apoptosis-deficient cells. Here, we provide further evidence to support the role of autophagy in cell killing after PDT. Autophagy was blocked by knockdown of one essential factor, LC3 or Atg7, in MCF-7 cells. The cells were exposed to a range of doses of PDT sensitized by the phthalocyanine Pc 4; steps in autophagy were monitored by western blotting for LC3-II and by fluorescence microscopy for the uptake of monodansylcadaverine or for the distribution of transfected GFP-LC3; and overall cell death was monitored by MTT assay and by clonogenic assay. We find that blocking autophagy increased the survival of MCF-7 cells after PDT and increased the shoulder on the dose-response curve. In response to Pc 4-PDT, Atg7-deficient MCF-7 cells remained capable of robust accumulation of LC3-II, but were defective in comparison to Atg7(+) cells in the formation of autophagosomes. We conclude that apoptosis-deficient cells rely on autophagy for cell death after Pc 4-PDT and that the strong activation of LC3 maturation in response to PDT could occur even in cells with limited or no Atg7 expression.
Xue
Liang-Yan
LY
Department of Radiation Oncology, Case Western Reserve University School of Medicine, Cleveland, OH, USA.
Chiu
Song-Mao
SM
Oleinick
Nancy L
NL
D017209:Apoptosis / Q000502:physiology; D001343:Autophagy / Q000502:physiology; D000071193:Autophagy-Related Protein 7; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D045744:Cell Line, Tumor / Q000528:radiation effects*; D005260:Female; D006801:Humans; D007211:Indoles / Q000627:therapeutic use; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000627:therapeutic use; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D044764:Ubiquitin-Activating Enzymes / Q000172:deficiency*
D007211:Indoles; C477012:MAP1LC3A protein, human; D008869:Microtubule-Associated Proteins; D017319:Photosensitizing Agents; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; C107448:phthalocyanine Pc 4; C509349:ATG7 protein, human; D000071193:Autophagy-Related Protein 7; D044764:Ubiquitin-Activating Enzymes