RPUSD1 role in NSCLC - Biología molecular

Por:
- Isabella Ríos Pérez
- Samuel Rodriguez Duque

Estudiantes de la Universidad Pontificia Bolivariana - Programa Medicina - 3er semestre.

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Presented By :
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Isabella Ríos Pérez
Samuel Rodríguez Duque
01Universidad Pontificia Bolivariana
Faculty of Health Sciences – Medicine Program – Third Semester

2. Image taken from: www.spandidos-
publications.com/10.3892/mmr.2019.10121 ProjectAboutHome
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Pseudouridine
synthase (RNA
stability)INTRODUCTION
Promotes mRNA
translation
Most common lung
cancer; linked to
PI3K/AKT activation
RPUSD1
eIF4E
NSCLC
PI3K/AKT
Signaling pathway that
drives cancer cell
growthKey Concepts

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03INTRODUCTION Proposed mechanism of RPUSD1 in NSCLC Image taken from: Figure 7, Zhang J, et al. Int J Biol Macromol.
2025;245:127603. 7G: Proposed mechanism: RPUSD1
→ eIF4E → NBS1 → PI3K/AKT 7F: Increased Ki67 with RPUSD1
overexpression

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04GENERAL
OBJECTIVE To analyze the role of
RPUSD1 in enhancing the
expression of eIF4E through
the RluA catalytic domain,
activating the PI3K/AKT
signaling pathway, and
promoting cell proliferation
and invasion in non-small
cell lung cancer.

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05• Fundamento: Detecta proteínas en tejido con anticuerpos.
• NSCLC en parafina + anti-RPUSD1 → DAB.
• Intensidad (0–2) × porcentaje (1–4); ≥2 = alta expresión.
• Correlación con características clínicopatológicas. 2.2 Inmunohistoquímica (IHC)
MÉTODOS

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MÉTODOS • Fundamento: Mide niveles de ARNm.
• ARN → ADNc → amplificación SYBR Green.
• Normalización con GAPDH; método 2^-ΔΔCt.
• Cuantificar RPUSD1, eIF4E y otros genes. 2.5 PCR cuantitativa en tiempo
real (qRT-PCR)

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2.6 Western blot
MÉTODOS
Fundamento: Separación, transferencia e
identificación de proteínas específicas.
Separar proteínas por tamaño (SDS-PAGE)
Transferirlas a una membrana
Detectar específicamente la proteína de interés
mediante anticuerpos
Revelando la señal con quimioluminiscencia.

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2.7 Tinción por inmunofluorescencia
MÉTODOS
Fundamento: Detectar la localización y abundancia de una proteína
específica
Anticuerpo primario: detecta de forma específica la proteína de interés
Anticuerpo secundario: con un fluoróforo permite visualizarla bajo luz
apropiada
Al combinarse con la fluorescencia de DAPI, se puede observar la
localización de la proteína en relación con el núcleo celular

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FIG. 1A — INMUNOHISTOQUIMICA
RESULTADOS
FIG. 1C— WESTERN BLOT

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RESULTADOS
FIG 2H — WESTERN BLOT
FIG. 1F — LÁSER CONFOCAL

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RESULTADOS
FIG 3H — WESTERN BLOT
FIG 4C — WESTERN BLOT

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RESULTADOS
FIG 5D — WESTERN BLOT

13. AUTHOR What the author stated
Agreement with our
results
Y.C. Chen et al.
NBS1 directly binds to the ABD domain of
PI3K p110, activating PI3K and
downstream AKT signaling.
Agrees — RPUSD1 upregulates
NBS1, triggering the same
activation pathway.
M.H. Yang et
al. / B. Liu et al.
p85 binds to the ABD domain of p110 and
inhibits PI3K/AKT; NBS1 competes for this
site, removing the inhibition. p85
structural domains stabilize p110 and
regulate its kinase activity.
Agrees — RPUSD1-induced
NBS1 likely displaces p85,
releasing inhibition and
allowing activation.
M. Del Campo
et al. / S.
Grunberg et al.
Asp67 in the RluA domain is essential for
RPUSD1´s catalytic pseudouridylation
activity.
Partially agrees — mutants
lacking catalytic activity still
promoted malignancy in our
study.
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DISCUSSION

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CONCLUSION
2. eIF4E plays a
key role in
RPUSD1-mediated
processes 3. The molecular
mechanisms of
RPUSD1 are
complex and
multifaceted 1.RPUSD1
promotes lung
cancer cell
growth and
invasion

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15Mapa conceptual - Isabella Ríos Pérez
https://lucid.app/lucidspark/3deddd5a-20a5-49f6-8316-5bafb2e224d7/edit?
viewport_loc=-163%2C347%2C3024%2C1841%2C0_0&invitationId=inv_97fe1a6d-4f66-489e-bc38-e572dd24a44f

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16Mapa conceptual - Samuel Rodríguez Duque