HLTF promotes hepatocellular carcinoma progression Juan David Ballut 000483796.pdf
1. 01
Juan David Ballut Rodriguez, Estudiante III semestre
Universidad Pontificia Bolivariana
30 de enero / 2023
2. INTRODUCTION
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The Helicase-like Transcription Factor (HLTF) belongs to the SWI/SNF
family of proteins involved in chromatin remodeling. In addition to its
role in gene transcription, HLTF has been implicated in DNA repair,
which suggests that this protein acts as a tumor suppressor.
HELICASE-LIKE TRANSCRIPTION FACTOR (HLTF) AND IT´S
RELATION TO CARCINOGENESIS
HLTF has a low methylationrate in HCC.
Two transcript variants encoding the same protein have
been found for this gene.
DNA methylation
3. Chain of proteins in the cell that communicates a signal from a receptor on the
surface of the cell to the DNA in the nucleus of the cell. MAPK can activate a
transcription factor, such as Myc. This process is described in more detail below.
SRSF1 AND ERK/MAPK PATHWAY
SRSF1
SRSF1 is the gene that codes for ASF/SF2 (essential sequence
specific splicing factor)
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4. To provide data of the underlying mechanism of HLTF in HCC progression for the first time.
OBJECTIVE
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5. DNA sample
DNA Primers
Raw nucleotides
DNA polymerase
Technique used to amplify
specific segments of DNA
You need:
METHODS
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Tissues or cells were lysed with RIPA buffer supplemented with protease and
phosphatase inhibitors, and proteins were harvested. Then, the proteins were
separated on PAGE gels and electrotransferred onto NC membranes.
REAL-TIME QUANTITATIVE PCR (QRT‒
PCR)
Total RNA was extracted using
an RNA Miniprep Kit
WESTERN BLOTTING
The expression levels of
mRNAs and miRNAs were
normalized to those of GAPDH
and U6
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RNA extraction
Reverse transcription into cDNA
Adapted ligation
Amplification
Sequencing
RNA sequencing was completed by JLX MEDICAL SCIENCES (Shanghai, China).
Briefly, Huh7 cells transfected with shHLTF-1 or shNC were used to extract total
RNA for quality control.
UBIQUITINATION ASSAY
This pathway targets proteins to
the proteasome, which degrades
and recycles the substrates.
E1 Ub-activating enzyme
E2 Ub-conjugating enzyme
E3 Ub ligase
METHODS
RNA SEQUENCING
Key enzymes:
9. AUTHOR
Ariel Shimoni-
Sebag
Yuji
Masuda
Wenjing
Yong
TAKEAWAYS
"It has been confirmed that SRSF1 may
promote the activation of the ERK/MAPK
signaling pathway by increasing the levels
of B-RAF mRNA and protein".
The stability of SRSF1 protein was significantly reduced in
HLTF knockdown HCC cells, but the ubiquitination of SRSF1
was enhanced in the presence of MG132, while the result was
the opposite in HLTF-overexpressing cells. HLTF has been
reported to act as an E3 ubiquitin ligase to promote the
polyubiquitination of PCNA by forming a thioesterlinked Ub
chain, thus participating in postreplication DNA repair.
In our study, we found by a luciferase reporter assay
that miR-511-5p, as the upstream regulator of HLTF,
directly targets HLTF. It has been reported that miR-
511-5p expression is markedly reduced in gastric
and colorectal cancer and functions as a tumor
suppressor.
IN AGREEMENT DISAGREEMENT
DISCUSSION
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10. CONCLUSSIONS
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Inhibitors of the Mapk pathway and
HLTF such as miR-511-5p can be utilized
as a form of treatment not cure of hcc
patients.
MAPK pathways as a cellular proliferation
factor while studying it, helps us to find
out which genes and environmental
factors have a critical role during the
development of cancer, so there would be
the possibility of finding those causes
and possible treatments for diseases that
were initially considered untreatable.