LUIGI BOURLOT ELENA GALLO

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LUIGI BOURLOT ELENA GALLO

CANCER STEM CELLS Stem cells property: Self-renewal Tumorigenesis Tumor growth Differentiation Identify in many tipes of cancer: Cervical cancer Cell-surface markers: CD133 Pluripotency-associated proteins (PA): Oct4, Sox2 We will see that: Autophagy has a role in CSCs pluripotency mainteinace Autophagy regulates the proliferation and invasion of CSCs Homeostasis of PA is regulated in hESCs by Autophagy (a conserved lysosomal degradation pathway , generally induced by extracellular stress).

CSCs HAVE STAMINAL PROPERTIES Low adhesion suspended culture of HeLa and CasKi Formation of tumorspheres Immunofluorescence for the detection of Oct4 and stem cells marker CD133 Presence of stem cells markers. Higher expression in tumorspheres. CSCs HAVE STAMINAL PROPERTIES

Authophagy induction with HBSS buffer (starvation). Use of Acridine Orange staining for the detection of autophagosomes Western blot for the expression level of autophagic protein LC3 II and Beclin I Autophagy is higher in cancer stem cells than in the general cancer cells

Analysis of proliferation ( Real-time Cellular Analysis noninvasive electrical impedance) afeter HBSS treatment. Analysis of invasion ( invasion assay using a transwell filter) after HBSS treatment. Western blot for the detection of PA after HBSS treatment in adherent and tumorspheres cells. A B C Authophagy regulate the proliferation, invasion of CSCs and the expression level of PA

Autophagy regulates stemness: Analysis of proliferation and invasion after 3-MA autophagy inhibition treatment. Western blot analysis of PA proteins after 3-MA treatment. A B Autophagy regulates stemness: 3-MA treated cells decrease their proliferation , invasion and PA expression.

Discussion CSCs have a critical role in the tumor genesis, growth , invasion, methastasis, tumor relapse and drugs resistance. These results demostrate that autophagy regulates the stemness of cervical cancer stem cells Understanding the mechanisms through which authophagy regulates the maintainance of stem properties may be foundamental to develop new therapheutic approaches. In the future stem cells could be a preference target in cancer treatment.