Seminario de Biología Molécular,tercer semestre de medicina.Universidad Pontificia Bolivariana

1. ISABELA MURILLO MORENO
FACULTY OF HEALTH SCIENCES-UPB
MEDICINE
THIRD SEMESTER
2023

2. What is cancer?
THE CANCER IS A BROAD TERM USED TO REFER TO A GROUP OF DISEASES THAT CAN ORIGINATE IN
ALMOST ANY ORGAN OR TISSUE IN THE BODY WHEN ABNORMAL CELLS GROW UNCONTROLLABLY,
EXCEED THEIR NORMAL LIMITS AND INVADE ADJACENT PARTS OF THE BODY AND/OR SPREAD TO OTHER
ORGANS .
CANCER CAN START ANYWHERE IN THE HUMAN BODY, MADE UP OF TRILLIONS OF CELLS.

3. TUMOR CELLS NEED A LARGE AMOUNT OF ENERGY TO MAINTAIN RAPID CELL
PROLIFERATION; THEY ACTIVELY REPROGRAM APOPTOSIS TO BECOME RESISTANT TO
DRUGS.
Introduction
ORAL SQUAMOUS CELL CARCINOMA IS A COMMON TUMOR WITH A HIGH INCIDENCE IN DEVELOPING COUNTRIES.

4. Objetive
This study aimed to determine whether glucose transporter-1/3 (GLUT1/3) increased
expression could contribute to oral tumor severity.
This image shows an example of a keratinizing
squamous cell carcinoma of the oral cavity.

5. Methods
WESTERN BLOT (WB)
GLUT1/3 RT-PCR
Forty-eight tissue samples from OSCC patients
and AN (controls) were arbitrarily chosen to
determine the mRNA expression of GLUT1 and
GLUT3
The tissue was crushed to powder with LN2, mixed
thoroughly in RIPA (Radio immunoprecipitation assay)
buffer, centrifuged, supernatant isolated (used as
protein), and quanti- fied (OD at 595 nm).

6. IHC (Immunohistochemistry)
staining of GLUT1/3
Cell
culture,treatmentofcisplatin/GLUT1
inhibitors,andMTTassay
The EnVision Kit (DAKO, USA; Cat no: K8023) and DAB
(diaminobenzidine tetrachloride) was used for staining.
We have included a negative con- trol (BSA replacing
the 10Ab) and a positive control (known GLUT1/3
immunoreactivity) in each staining batch.
The experiments were performed in 6-well
plates. 5 μM of GLUT inhibitor (BAY-876,
SML1774, Sigma-Aldrich) was treated for 12
hours. After that 20 μM cisplatin/vehicle
control was treated, and the cells were
harvested after 24 hours.

7. Results
Fig. 2. GLUT3 protein and mRNA expression in primary and
drug-resistant oral tumor samples. (A) Representative
Western Blot and RT-PCR gel showing the expression of
GLUT3 protein in AN, PT, N1-3 and RCRT specimens as
indicated.
Fig. 1. GLUT1 protein and mRNA expression in primary and drug-resistant oral
tumor samples. (A) Representative Western Blot and RT-PCR gel showing the
expression of GLUT1 protein in tumor-adjacent (AN) normal, primary tumor (PT),
invasive (N1-3) and chemo-radiation therapy-resistant recurrent tumors (RCRT)
as indicated. The dot plot with error bars of GLUT1

8. Fig. 3. GLUT1/3 protein expression in primary and drug-resistant oral tumor samples. (A–D) Representative
IHC staining of oral cancer (OSCC; A-AN, B-PT, C- N1-3 and D-RCRT) showed varying IR to GLUT1 antibody.
(E–H) Representative IHC staining of oral cancer (OSCC; A-AN, B-PT, C-N1-3 and D-RCRT) samples showing
varying immunoreactivity to GLUT3 antibody.
Fig. 4. Expression of OTFs like HIF1α, AP1 (c-Jun, c-Fos and Fra-2), and NFκB in OSCC and their
correlation with GLUT1/3 mRNA/Protein expression. (A) HIF1α, AP1 (c-Jun, Fra-2 and c-Fos), and NFκB
protein expression at various stages of OSCC progression.

9. GLUT1/3 protein expressionincreases with histological
grade/stage, lymph node metastasis, and distant
metastasis of OSCC
GLUT1 WAS UPREGULATED IN HUMAN OSCC-DERIVED CELLS
UNDER HYPOXIA
Discussion
author submits that match?
(Azad et al., 2016; Feitosa et al., 2018; Li
et al., 2016).
BCL-2 INHIBITOR-INDUCED BECLIN-1/ATG5 MEDIATED AUTOPHAGY
(Liang et al., 2015).
􏰪􏰨􏰕(Shimanishi et al., 2013)􏰄􏰞􏰊􏰆􏰄􏰃􏰕􏰄 􏰁􏰟 􏰊􏰈􏰐􏰠 􏰗􏰏􏰎􏰘􏰬􏰐

10. Conclusions
WHAT WOULD BE OF MEDICINE?
WITHOUT THE POWER OF
SPECIFIC DETECTION AND
QUANTIFICATION OF GENETIC
MATERIAL IN A BIOLOGICAL
SAMPLE.
THIS AREA HAS SHOWN A
SIGNIFICANT IMPACT ON HEALTH,
AS IN THIS TYPE OF INVASIVE
CANCER, WHERE IT ALLOWS US
TO FIND IN A COMPREHENSIVE
WAY AND BEYOND WHAT WE CAN
SEE WHERE THERE IS AN
ALTERATION AND HOW
THERAPEUTICALLY WE CAN FIND
A SOLUTION THAT REVOLVES
AROUND TO THE WELL-BEING OF
PATIENTS.
MOLECULAR BIOLOGY
IS ESSENTIAL IN
MEDICINE TO
UNDERSTAND
FUNDAMENTAL
BIOLOGICAL
PROCESSES SUCH AS
CELL FUNCTIONS AND
BEHAVIORS IN
CERTAIN SITUATIONS
FOR THE TREATMENT
OF DIFFERENT
DISEASES.

11. Thank
Thank
Thank
you!
you!
you!