Seminario biología molecular, III semestre medicina. Universidad Pontificia Bolivariana.

1. María José Pérez Zabala
III semestre
Universidad Pontificia Bolivariana
Medellín, Colombia

2. INTRODUCTION
PROTEOME
“Is the complete set of proteins made by an organism”
Also, the changes that this one haves differs from one cell to
another (different part of the bodie) and dependes also on what
stage of life the individual is and environmental conditions. RELEVANT PART
Important fact
Information about the proteome
helps to find out which proteins are
involved in specific diseases, also it
helps to develop medicines.

3. INTRODUCTION
4-(methylnitrosamin)-1-(3-
piridyl)-1-butanone or
Nicotine-derived nitrosone (NNK)
is one of the nitrosamines
derived from the nicotine in
tabaco
4-(methylnitrosamin)-1-(3- piridyl)-1-butanone
What is it?

4. How proteome derangement in malignant
epithelial cells and its stroma are related
to the exposure to 4‐(methylnitrosamino)‐
1‐(3‐pyridyl)‐1‐butanone
GENERAL OBJECTIVE

5. METHODS
2D-DIGE Immunoblotting
Basis:
Electrophoresis is performed with coomassie
brilliant blue and polyacrylamide gel. Sodium
Duodecyl Sulfate - Polyacrylamide Gel
Electrophoresis is the most popular method
for determining polypeptide size in the
presence of SDS, since a known size standard
is used and mobility is determined by the
molecular weight and electrical charge of the
molecule.
Basis:
It is another variation of the Southern
transfer. Proteins from cell extracts are
separated by gel eletrophoresis according to
their size
After that the proteins are transferred to a
filter, which is incubated with antibodies that
react with the protein of interest. The
antibody bound to the filter can be detected in
several ways, thus identifying the protein
against which the antibody is directed.
Why did they do it? Why did they do it?
Were used to confirm the protein
changes in animal tissues as observed
in 2D-DIGE and probe for the protein
changes in NNK transformed BEAS- 2B
cells. In addition, traditional Western
blot was used to measure the protein
changes of interest.
Protein identification.

6. METHODS
qRT‐PCR Immunohistochemistry
Basis:
It is the RNA amplification through its
cDNA synthesized a priori, using a reverse
transcriptase, followed by several cycles
of conceptional PCR. This technique can
determine the expression of genes in
various tissues .
Basis:
Is the identificacion of a tissue by
means of a specific interaction between
an antigen and a antibody has been
visibly labeled, the cell is colored to
demonstrate the presence and cellular
localization of a molecule of interest.
Why did they do it? Why did they do it?
Was used for amplification of total
RNA, E-cadherin, TWIST1, vimentin,
GAPDH
IHC for E-cadherin, vimentin, and
HRF were examined to substantiate
findings from Western blotting.
Slides were compared for
qualitative changes in protein
expression.

7. RESULTS

8. RESULTS

9. DISCUSSION
DISAGREE

10. CONCLUSIONS
Molecular biology techniques helps us to confirm the
expression or not of many proteins in a certain
pathology like it was for cancer of the lung in the
article.
Learning about molecular biology techniques
(Immunoblotting/ PCR/ etc) give us another perspective
of what is really happing with the patient or with
analyzed subject, gives us the precision we need for
treating correctly our patient.