Seminario biología molecular, tercer semestre medicina, Universidad pontificia Bolivariana

1. SEBASTIÁN HERNÁNDEZ CANO
3RD SEMESTER
MOLECULAR BIOLOGY
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2. Breast cancer
Reffers to a pathogenic growth of
cells around the breast area that
lose the ability to die
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3. KDM4Asignaling
pathway
Induces cell growth,
proliferation and survival
by regulating (+) DNA
repair processes
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HTTPS://WWW.GOOGLE.COM/URL?
SA=I&URL=HTTPS%3A%2F%2FLINK.SPRINGER.COM%2FARTICLE%2F10.1007%2FS00018-
019-03289-
W&PSIG=AOVVAW33FJG0_CETROGROUJ6JK2I&UST=1675647277961000&SOURCE=IMAG
ES&CD=VFE&VED=0CBAQJRXQFWOTCNIX9NEE_FWCFQAAAAADAAAAABAE

4. General Objective
General Objective
"Evaluate epigenetic regulator KDM4A activates
Notch1-NICD-dependent signaling to drive
tumorigenesis and metastasis in breast cancer "
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5. Materials and Methods
INMUNOHISTOCHEMICAL STAINING
RNA ISOLATION AND
QUANTITATIVE RT-PCR
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50 samples were extracted and
analyzed with GT vision III staining kit
Then, staining was classified as none
(0), weak (1), moderate (2) and
strong (3) both by intensity and
covered area
mRNA was isolated and transcribed
to cDNA by reverse transcriptase,
then PCR was performed (RT-PCR
process)

6. inmmunoreactions antigen-antibody were
performed for KDM4A, Notch1, CCND3
and β-actin.
gels were performed using electrophoresis
Materialsandmethods
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Western Blot analysis
Proteins of interest bound to DNA were
marked with antibodies (H3K27ac and
H3K9me3). the DNA was then purified and
qPCR was performed
ChIP-qPCR assay
https://www.google.com/url?sa=i&url=https%3A%2F%2Fwww.bio-rad.com%2Fen-us%2Fapplications-technologies%2Fprotein-electrophoresis-
methods&psig=AOvVaw2eMLzREzOM6SyYWpkiPaEG&ust=1675639707073000&source=images&cd=vfe&ved=0CBAQjRxqFwoTCKi5vr6C_fwCFQA
AAAAdAAAAABAE
https://www.google.com/url?sa=i&url=https%3A%2F%2Fwww.cellsignal.com%2Fapplications%2Fchip-and-
chip-seq%2Fregulation-expression-in-cell-and-
tissue&psig=AOvVaw2ZZtAvhnjdjAtJX9TlhYA2&ust=1675641402433000&source=images&cd=vfe&ved=0
CBAQjRxqFwoTCNjBheaI_fwCFQAAAAAdAAAAABAE

7. KDM4A
SILENCER RNA NOTCH1 SIGNALING
IN KNOCKDOWNS
LOW/HIGH
CLASSIFICATION
Results
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Low High

8. KDM4A KNOCKDOWN (F)
AND OVEREXPRESSION (G)
KDM4A INCREASES CELL
VIABILITY AND
PROLIFERATION
NOTCH 1 INHIBITION COULD
SUPRESS DOWNSTREAM
GENES
Results
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9. Author Statement Status
S.Z. Cui
In prostate cancer, USP1 is a deubiquitinase
that regulates KDM4A K48-linked deubiquitin
and stability to drive tumor cells proliferation
and enzalutamide resistance
Agree
J. Lin
Oncogene APOL1 promotes proliferation and
inhibits apoptosis via activating NOTCH1
signaling pathway in pancreatic cancer
Agree
M.A.J. Morgan, A. Shilatifard
histone modifications exert essential functions
in cell fate determination, terminal
differentiation and X inactivation
Agree
Discussion
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10. Conclusions
Asserting molecular pathways in pathological
events could help provide therapeutic targets such
as KDM4A or Notch1
learning about the tissue behavior behind protein
overexpression/knockdown might help establish
the reason to certain pathologies and develop more
specific and efficient treatments
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11. Epigenetic regulator KDM4A activates Notch1-
NICD-dependent signaling to drive tumorigenesis
and metastasis in breast cancer
NOTCH1
KDM4A is a histone demethylase
that works in DNA repair,
KDM4A
stimulates
An unusual and uncontrolled cell
proliferation that leads to
tumorgenesis
cell growth and proliferation
Overexpression
neoplasic events as well as
metastasis or complications related
to tumor resilience and spreading
speed
Lower survival outcomes
Inhibiting th pathway and its downstream proteins could
help reduce tumor viability, making it easier to deal with it
as well as reducing metastasis probabilities
Knockdown
Higher survival outcomes
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