Cholesterol homeostasis and autophagic flux in perifosine-treated human hepatoblastoma HepG2 and glioblastoma U-87 MG cell lines.
Perifosine exerts an antiproliferative effect on HepG2 and U-87 MG cells and also interferes with the transport of cholesterol from the plasma membrane to the endoplasmic reticulum (ER). Recently we demonstrated that exposure of U-87 MG cells to perifosine causes an accumulation of autophagosomes. We have now expanded the study to establish the molecular mechanism by which perifosine interferes with the autophagic process. Using transmission electron microscopy, we report that the treatment of HepG2 and U-87 MG cells with perifosine causes an intense cytoplasmic vacuolization identified as autophagic vesicles. The accumulation of autophagosomes induced by perifosine is due to a blockage of the autophagic flux, thereby affecting cell proliferation. Perifosine also provokes a differential ER stress response in the HepG2 and U-87 MG cell lines. We have also demonstrated a relationship between the deregulation of cholesterol transport and the inhibition of the autophagic flux prompted by perifosine. Thus our findings clearly demonstrate that perifosine impairs the autophagic flux in HepG2 and U-87 MG cells, which is related to defects in intracellular cholesterol transport. Our study is relevant for anticancer therapy because tumour cells exhibit autophagy as a pro-survival mechanism. Further research to identify the precise mechanisms of autophagy maturation and the role of cholesterol may provide new insights into the antiproliferative action of perifosine.
Department of Biochemistry and Molecular Biology I, Faculty of Sciences, University of Granada, Av. Fuentenueva s/n, Granada 18071, Spain(2).
Pablo
P
Ríos-Marco
Department of Cell Biology, Faculty of Sciences, University of Granada, Av. Fuentenueva s/n, Granada 18071, Spain(3).
Antonio
A
Ríos
Department of Biochemistry and Molecular Biology I, Faculty of Sciences, University of Granada, Av. Fuentenueva s/n, Granada 18071, Spain(2).
José M
JM
Jiménez-López
Department of Biochemistry and Molecular Biology I, Faculty of Sciences, University of Granada, Av. Fuentenueva s/n, Granada 18071, Spain(2). Electronic address: mpazcj@ugr.es.
María P
MP
Carrasco
Department of Biochemistry and Molecular Biology I, Faculty of Sciences, University of Granada, Av. Fuentenueva s/n, Granada 18071, Spain(2). Electronic address: cmarco@ugr.es.
Carmen
C
Marco
D001343:Autophagy; D001932:Brain Neoplasms / Q000473:pathology; D045744:Cell Line, Tumor; D002784:Cholesterol / Q000378:metabolism*; D005909:Glioblastoma / Q000473:pathology*; D018197:Hepatoblastoma / Q000473:pathology*; D006706:Homeostasis; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology
D010767:Phosphorylcholine; C105905:perifosine; D002784:Cholesterol
Antitumour agents; Autophagy; Cholesterol homeostasis; HepG2 and U-87 MG cells; Perifosine