5-aminolevulinic acid photodynamic therapy reduces HPV viral load via autophagy and apoptosis by modulating Ras/Raf/MEK/ERK and PI3K/AKT pathways in HeLa cells

J Photochem Photobiol B. 2019 May:194:46-55. doi: 10.1016/j.jphotobiol.2019.03.012. Epub 2019 Mar 21.

Abstract

Human papillomavirus (HPV) infection is linked to several diseases, the most prominent of which are cervical cancer and genital condyloma acuminatum. Previous studies have suggested an effective role for 5-aminolevulinic acid photodynamic therapy (ALA-PDT) against various cancers by the induction of autophagy and apoptosis. However, few reports have focused on the effectiveness of ALA-PDT on HPV related disorders. To identify the role of ALA-PDT in the context of HPV infection, we initially investigated 111 patients suffering from genital condyloma acuminatum. HPV viral load detected before and after ALA-PDT treatment was compared during this procedure; a significant difference was noted. HeLa (HPV18) cells were exposed to ALA-PDT in vitro to further explore the underlying mechanisms. Western blot analysis showed that ALA-PDT induces LC3II and p62 expression, along with the up regulation of caspase-3 and cleaved caspase-3. Our study also demonstrated that ALA-PDT treatment inhibits the proliferation of HeLa cells in a dose dependent manner and effectively reduces HPV viral load via autophagy and apoptosis by regulating the Ras/Raf/MEK/ERK and PI3K/AKT/mTOR pathways. Hydroxychloroquine (HCQ), although it inhibited autophagy degradation, functioned to activate reactive oxygen species (ROS) levels of ALA-PDT to enhance the observed effect. These findings suggest strategies for the improvement of PDT efficacy in patients.

Keywords: ALA-PDT; Apoptosis; Autophagy; HPV; HeLa.

MeSH terms

  • Aminolevulinic Acid
  • Apoptosis / drug effects
  • Apoptosis / radiation effects
  • Autophagy / drug effects
  • Autophagy / radiation effects
  • Cell Death / drug effects*
  • Cell Death / radiation effects*
  • Cell Proliferation / drug effects
  • Cell Proliferation / radiation effects
  • Cell Survival / drug effects
  • Cell Survival / radiation effects
  • Dose-Response Relationship, Drug
  • Dose-Response Relationship, Radiation
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • HeLa Cells
  • Humans
  • Levulinic Acids / pharmacology*
  • MAP Kinase Signaling System / drug effects
  • MAP Kinase Signaling System / radiation effects
  • Papillomaviridae / drug effects
  • Papillomaviridae / physiology*
  • Papillomaviridae / radiation effects
  • Phosphatidylinositol 3-Kinases / metabolism
  • Photochemotherapy*
  • Proto-Oncogene Proteins c-akt / metabolism
  • Reactive Oxygen Species / metabolism
  • TOR Serine-Threonine Kinases / metabolism
  • Viral Load / drug effects*
  • Viral Load / radiation effects*
  • raf Kinases / metabolism
  • ras Proteins / metabolism

Substances

  • Levulinic Acids
  • Reactive Oxygen Species
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • raf Kinases
  • Extracellular Signal-Regulated MAP Kinases
  • ras Proteins