Seminario biología molecular Universidad pontifica bolivariana Tercer semestre

1. Paulina Morales Rodriguez
III semestre

2. HEPATOCELLULAR
CARCINOMA
Is the most common type of primary liver
cancer in adults. Representing the 6th
most commonly diagnosed type of cancer
with a percentage of 75% to 85% of sick
people.

3. Contributor tumor
progression is given by:
Increasing evidences showed that
m RNA were frequently abnormally
expressed in tumor tissues.
Suggesting that these molecules
had important roles in tumor
progression and metastasis.
The microRNAs are a class of small
endogenous non-coding RNAs
that are typically between gene
expression post-transcriptionally
The tumor microenvironment, tumor
cells, innate and adaptive immune
cells, cancer associated fibroblast.
The m RNA translational inhibition:

4. OBJETIVES
53 M
Explore the role and mechanisms
of cancer associated fibroblasts
exosomes in promoting
hepatocellular cancer progression
with:
Suggested that cancer
associated fibroblasts
exosomes, promoted the
aggressive phenotype of
hepatocellular cancer cells
Imprinted region are
involved in the epithelial-
mesenchymal transition
process and oncogenesis

5. METHODS
CELL CULTURE AND
TRANSFECTION
IMMUNOFLUORESCENCE
This cells were cultured in high-glucose,
dulbeccos modified eagles medium,
supplemented with 10% fetal bovine serum, 1%
penicillin an 100 mirogr streotomycin at 37 C
with 5% of CO2. m RNA mimics were design
and synthesized by genepharma and were
transfected into hepatocellular cancer cells
Cells were fixed in 4% paraformaldehyde and
blocked with 5% bovine serum albumin. Next,
cells were incubated with anti alfa- SMA and
anti-vitamentin at 4C for a night, followed by
incubation with 488 conjugated secondary
antibody for 2 hours. After washing with PBS,
cells were counterstained with DAPI for
observation under the microscope

6. METHODS
WESTERN BLOT RNA EXTRACTION AND PCR
Was carried out using standard procedures as
previously descried. The antibodies used for
western blot assay were as follows: anti alfa
SMA, anti-vimentin, TSG101, CD8, CD9, E-
cadherin, N-cadhern, HHIP, and GAPDH. The
protein level was standardized to GAPDH, and
the standardized to experimental control.
Total RNA of cells was extracted by the
RNA isolation kits. For m RNA detection, c
DNA was synthesized by the quantitect
reverse transcription kit. All operations were
carried out according to manufacturers
instructions. The m RNA expression levels
of gene were quantified by SYBR mix and
roche real time PCR detection system

7. RESULTS
1 2 3

8. RESULTS
4 5 6

9. CONCLUSION
AUTHOR
Exosomes secreted by tumor cells
can modulate varius biological
processes in tumor progression,
including tumor angiogenesis,
metastasis and immune escape
Wang M
Chang C
Tada M
OPINION
The DLK1-DIO3 miRNA cluster
plays a significant role in cancer
self renerwal and maintenance
of the aggressive phenotype
A previuous study reported that
DNA hypermethylation and/or loss
oh HHIP transcription, and Hh
signal activation through the
activation oh HHIP was impicated
in the pathogenesis oh HCC

10. CONCLUSIONS
The molecular part of this experiment is fundamental since
thanks to these bases it is possible to understand and
analyze how the interactions of the different systems of the
cell occur, which includes many relationships, the synthesis
of proteins, metabolism, and how all these interactions are
regulated to achieve proper cell function.
Allow the study of the DNA of human cells (their complete
genome or only the genes potentially involved in the disease),
to determine the type of alteration and/or cancer that it
presents, its prognosis and/or therapeutic alternatives to
follow.