Seminario biología molecular - UPB (3 semestre)

En este seminario hablamos sobre el cáncer de mama

1. Estudiantes:
Manuela Betancur Toro
Santiago Bonilla
Tercer semestre
SEMINARIO

2. BREAST CANCER
It is a disease in which
altered breast cells multiply
uncontrollably. The cancerous
cells begin to develop within
the milk ducts or milk-
producing lobules and can
metastasize by spreading to
other organs. Causing a high
mortality rate worldwide.
INTRODUCTION

3. miRNA-499-5pSOX6
SOX6 is a member of the SoxD
subfamily of transcription factors. It
is involved in organ development and
various physiological processes.
In this study, suppression of SOX6
induced EMT in 4T1 breast cancer
cells and activated downstream
pathways related to EMT.
Is a microRNA that can regulate
gene expression.
Previous studies have shown that
miR-499-5p can promote invasion
and EMT in some types of cancer,
such as lung and colon cancer.
This study found that propranolol
treatment decreased miR-499-5p
expression in 4T1 breast cancer
cells.
INTRODUCTION

4. This study will focus on 4T1 cells, a murine
mammary adenocarcinoma cell line, as the
primary research subject.
They investigated the inhibitory efects and
mechanisms of propranolol on epithelial–
mesenchymal transition (EMT) in breast
cancer cells, aiming to elucidate this
phenomenon at the miRNA level
GENERAL OBJECTIVE

5. METHODS
La secuenciación de ADN es una técnica
utilizada para determinar el orden de los
nucleótidos (adenina, guanina, citosina y
timina) en una molécula de ADN.
En este estudio, la secuenciación permite
determinar la secuencia exacta de nucleótidos
en los microARN (miRNA) y los genes.
SECUENCIACION

6. RT-PCR
METHODS
Se utiliza para la detección y amplificación del ARN.
Permite estudiar la expresión de determinados
genes (su traducción en proteínas). En primer lugar,
se extrae el ARN total de las células estudiadas y se
separa la fracción correspondiente al mensajero
(ARNm).
En este estudio se utilizó para validar los cambios en
la expresión de miRNAs específicos.

7. WESTERN BLOT
METHODS
Permite la detección de una única proteína dentro de
una muestra biológica. La especificidad del Western
Blot se consigue utilizando un anticuerpo que reconoce
la proteína de interés.. El método implica el uso de
electroforesis en gel para separar las proteínas de la
muestra.
EN este estudio se utilizó para medir los cambios en la
expresión de proteínas relacionadas con EMT como E-
cadherina y vimentina.

8. INMUNOHISTOQUIMICA
METHODS
Es un procedimiento histopatológico basado en
la utilización de anticuerpos que, mediante
reacciones antígeno-anticuerpo, posteriormente
reveladas enzimáticamente, permiten la
identificación de marcadores antigénicos en los
tejidos, que se observan al microscopio óptico.
Se uso para visualizar los cambios en marcadores
de EMT in vivo e in vitro, en moleculas de cancer
de mama.

9. Figure 1
RESULTS
WESTERN BLOTINMUNOHISTOQUIMICA
WESTERN BLOT
INMUNOFLUORESCENCIA

10. Figure 2
RESULTS
WESTERN BLOT

11. Figure 3
RESULTS
WESTERN BLOT
RT-PCR

12. Figure 4
RESULTS
WESTERN BLOT

13. AUTHOR ARGUMENT AGREE/DESAGREE
Nieto, et al.
While EMT is closely related to cell differentiation, embryonic development,
and tissue repair, it also plays a pivotal role in tumor metastasis
He, et al.
Liu, et al.
Furthermore, it has been proven that miR-499-5p can promote the invasion
and EMT in lung and colon cancer cells
Canel, et al.
E-cadherin is an essential component of the cytoskeleton and intercellular
adhesion
DISCUSION

14. As we know, propranolol, a drug
traditionally used to treat
cardiovascular diseases, shows
promising potential as an
anticancer treatment, especially
in the prevention of metastasis.
Its ability to inhibit epithelial-
mesenchymal transition (EMT) in
breast cancer cells could open
new therapeutic avenues for
patients with metastatic cancer.
The results show how manipulation of
specific molecular factors (miR-499-
5p, Sox6) can alter cell phenotype,
evidenced by changes in EMT markers
such as E-cadherin and vimentin. This
highlights the plasticity of cancer
cells and the importance of
understanding the molecular
mechanisms that control it.
The identification of Sox6 as a
direct target of miR-499-5p
illustrates the specificity of
miRNA-mRNA interactions and
how these can influence broader
signaling cascades. This finding
reinforces the importance of
studying these interactions in
detail to understand gene
regulation.
3.2.1.
CONCLUSIONS

15. CONCEPTUAL MAP

16. CONCEPTUAL MAP