1. LUIGI BOURLOT ELENA GALLO

2. 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).

3. 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

4. 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

5. 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

6. 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.

7. 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.