Seminario Biología Molecular Tercer Semestre Medicina Universidad Pontificia Bolivariana

1. JUAN DIEGO GÓMEZ
AGUDELO
TERCER SEMESTRE
Universidad Pontificia Bolivariana

2. RHEUMATOID
ARTHRITIS
RA is a systemic autoimmune disease
characterized by synovial dysplasia and
migration of synovial fibroblasts, resulting in
erosion of cartilage and bone, and leading to
joint destruction and dysfunction.
During the pathogenesis of RA, various
inflammatory cells such as T cells, B cells, and
macrophages are activated to release
inflammatory factors to stimulate the activation
of synovial fibroblasts, leading to the increase
of synovial fibroblasts proliferation and
migration.
2

3. MARKERS
TRIM 33
It is thought to be a transcriptional
corepressor.
They interact with transcriptional factor to
repress the transcription of certain genes
binding. A correpresor binds directly to a
repressor to control DNA transcription.
KLF 9
It is a transcription factor that binds to GC
box elements located in the promoter.
Binding of the encoded protein to a single
GC box inhibits mRNA expression while
binding to tandemly repeated GC box
elements activates transcription.

4. OBJECTIVE
This study aimed to investigate how
TRIM33 regulated by KLF9 affects the
aggressive behaviors of synovial
fibroblasts induced by tumor necrosis
factor‐α (TNF‐α).

5. METHODS
Polymerase chain reaction (abbreviated PCR) is a
laboratory technique for rapidly producing
(amplifying) millions to billions of copies of a specific
segment of DNA, which can then be studied in
greater detail.
Western blot
PCR
Western blotting is a laboratory technique used to
detect a specific protein in a blood or tissue sample.
• Gel electrophoresis to separate the sample's
proteins.
• The membrane is exposed to an antibody specific
to the target protein.
Western blot tested the expression of
metastasis related proteins and many other
molecules.
To test TRIM33 expression in control
and TNF

6. METHODS
It is based on covalently linking an enzyme to a known
antigen or antibody, reacting the enzyme-linked material
with the patient’s specimen, and then assaying for
enzyme activity by adding the substrate of the enzyme.
Enzyme activity is measured by adding the substrate for
the enzyme and estimating the color reaction in a
spectrophotometer.
ELISA Dual-Luciferase Reporter Assay
Luciferase assays can be used to investigate a
variety of cellular processes using bioluminescent
light output as a sensitive measure of biological
activity.
Different IL levels were measured with ELISA, to see
how it would behave over an overexpression of
TRIM33.
Dual – luciferase reporter assay reported TRIM33
MUT and WT by the expression of specific RNA of
KLF9 and the control.

7. RESULTS

8. RESULTS

9. DISCUSSION
Author What they said Agreed/Disagreed
Kulkarni A. “Recent studies have found
that TRIM33 gene is lowly
expressed in liver cancer and
chronic myelogenous
Leukemia”
Agreed
Qi G. “In vitro and in vivo studies have
confirmed that TRIM33 plays an
important role in valproic acid
inhibiting the migration and
invasion of prostate cancer cells”
Agreed
Li L. and Chen M. “Previous studies hinted that KLF9
suppressed TNF‐α‐induced MH7A
cells or RA synovial fibroblast
inflammation, migration, and
proliferation”
Agreed

10. CONCLUSIONS
• Understanding the molecular composition
allows us to study biological processes and
their alterations in more depth, thus
allowing us to search for medical
treatments by experimenting on cellular
components in order to reverse or alter
physiological or pathophysiological
mechanisms.
• Molecular biology allows to apply methods that
allow to know values and reactions of different
components, showing their action within different
organisms including the human, thus achieving
studies and experiments to achieve evolutionary
processes that can become beneficial.