Hecho por: Samuel Salazar Arango Laura Camila Sepulveda

1. Laura Camila Sepúlveda Rivas - Samuel Salazar Arango
3rd semester, Medicine
Universidad Pontificia Bolivariana

2. INTRODUCTION
P53
Is the guardian of the genome, plays a
crucial role in maintaining the checkpoint
and passage of cells into the next phase.
→ altered p53 activity can lead to either
arrested cell proliferation or unregulated
hyperplasia.
→ Moreover, a clear involvement of p53 in
cell cycle re-entry,a common pathway in
apoptosis in neurodegeneration, has been
shown

3. INTRODUCTION
CELL CYCLE
Cell cycle is the name we give the
process through which cells replicate
and make two new cells. Cell cycle has
different stages called G1, S, G2, and
M. G1 is the stage where the cell is
preparing to divide. To do this, it then
moves into the S phase where the cell
copies all the DNA, then in the G2 the
cell growth and produce proteins, the
in the M phase the cell divide

4. NEUROBLASTOMA
→ Neuroblastoma is a malignant blue-cell tumor made up of neuroblasts.
Their growth is fast and they frequently have areas of bleeding,
necrosisand and calcification.
→ Is the most common malignant tumor in the first year of life
→ Neuroblastoma accounts for approximately 8% of all solid tumors in
childhood, and yet it assumes 15% of mortality related to solid tumors.

5. OBJECTIVE

6. Methodology

7. Western blot analyses
→ Is a laboratory technique used to detect a specific
protein in a blood or tissue sample using primary
antibodies directed against the protein in question
→ This study is evaluating the presence of proteins
- ADAM10
- ADAM17
- BACE1
- βAPP
- p53
- Calpain 2
- β-actin

8. Real-time quantitative polymerase
chain reaction
Is a PCR-based technique that
couples amplification of a target
DNA sequence with quantification
of the concentration of that DNA
species in the reaction.
Total RNA was extracted and
purified
Real-time PCR was performed
with 100 ng of total RNA
using the QuantiFast SYBR
Green RT-PCR kit

9. Cell viability assay
→ This measures cell viability in real-time.
→ In this assay, an engineered substrate is added directly to the corp medium.
→ The prosubstrate can penetrate cell membranes and enter cells. However, only
viable cells with active metabolism can reduce the prosubstrate into a substrate.
→ Formazan crystals formation is then checked under a microscope and the
media is replaced by 100 % DMSO until the purple crystals dissolved.

10. What is a flow cytometry
Flow cytometry has been used to identify distinct cell types within a
heterogeneous pool of cells, based on extracellular or surface marker
expression, an application commonly known as immunophenotyping
→ cells were trypsinized and dispersed into single cells.
→ All treated cells were collected and washed with ice cold.
Washed cells were fixed and immobilized in ice cold 70 % ethanol
until required.
→ Stained cells were analysed using a FACS Calibur flow
cytometer
Sub G1 and cell cycle analysis by
flow cytometer

11. RESULTS

12. RESULTS

13. RESULTS

14. RESULTS

15. Discussion
AUTHORS What did he say? Agree?
Long and Holtzman Considering the multifactorial and complex nature of AD, multidimensional
therapeutic strategies are currently adopted to prevent or slow disease
progression rather than cure the disease, which often seems to be impractical
Ittner and Ittner The seminal molecular event is an unbalanced processing of the amyloid
precursor protein, initiating a mechanism bringing into play synergistically
toxic amyloid peptides and the hyperphosphorylated forms of Tau
Vincent and Checler Its propensity to chelate zinc gives it the ability to inhibit the
metalloproteases ADAM10 and ADAM17, which are responsible for the non-
amyloidogenic processing of βAPP
- Dizdaroglu
- Mazumder
Phe was previously reported to inhibit transcription by interfering with
metal-containing DNA binding domain of specific proteins

16. 1 2 3
CONCLUSIONS
Thanks to molecular biology studies, it
was determined that antimicrobial and
anti-cancer therapies might trigger
Alzheimer's disease pathogenesis, which is
essential for its prevention
Studying the consequences of α-
secretase inhibition on p53 status in
cultured human neuroblastoma, we
could advance in treatment
The simultaneous targeting of βAPP
processing and p53 levels could serve
as a therapeutic approach since it
would alleviate multiple pathological
processes leading to neuronal
vulnerability and death.

17. CREDITS: This presentation template was created
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THANKS
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