2. CYCLIN D1
• What is it?
It’s a protein that regulates the progression through the G1 phase and
the G1 to S phase transition of the cell cycle.
• Function
It regulates the cell cycle by altering the transcription of genes that
control adhesion and migration.
• Reactions
• ⇧cell adhesion to stromal cells and fibronectin.
• Stabilizes F-actin fiber.
• Enhances chemotaxis and inflammatory chemokine secretion.
• Disrupts the redox balance by producing reactive oxygen species.
• Important for tumor initiation, maintenance, progression and
metastasis.
3. REDOX STATUS
• What is it?
It’s the balance between oxidants (or pro-oxidants)
and antioxidants.
Oxidants, including free radicals and other reactive
species, are continuously produced in the cell.
As it is impossible to completely prevent oxidant
production, several antioxidant systems have evolved
in the cell. In order to maintain a healthy status,
oxidants and antioxidants should be in equilibrium.
• Interactions
Oxidation is loss of electrons.
Reduction is gain of electrons.
4. ADHESION AND MIGRATION
→ ADHESION: the process of cells
assembling together in organized
and distinctive patterns to a surface
and, along with their connections to
the internal cytoskeleton through
cell adhesion molecules, it
determines the overall architecture
of the tissue.
→ MIGRATION: movement of cells of
different lineages over short and
long distances throughout the body
mainly during embryonic stage,
wound healing and immune
responses.
5. DRUG RESISTANCE
• What is it?
The reduction in effectiveness of a drug such as
antimicrobial, anthelmintic or antineoplastic in curing
a disease or condition, it’s a consequence of
evolution.
→ Inherent properties, such as genetic characteristics.
→ Acquired resistance after drug exposure.
6. MULTIPLE MYELOMA
• What is it?
It’s an incurable hemopathy
characterized by the
accumulation of clonal plasma
cells within the bone marrow
and the overproduction of
monoclonal immunoglobulin.
Clinical signs:
• Recurrent infections
• Immunodeficiency
• Renal failure
• Bone lesions
7. Bone
Marrow
Cells
Cellular
cycle
G1 to S
Phase
By Cycling
D1
Redox
unbalance
☝Cell
adhesión
and
migration
Stabilize F
actin fiber
Enhance
chemotaxis
and
inflammation
TUMOR:
initiation,
maintenance,
progression,
metastasis
Multiple
Myeloma
IT ALL DEPENDS
ON DRUG
RESISTENCE
8. OBJECTIVE
The objective of this article is to uncover the
functional implication of cyclin D1 in cells
microenvironment and in the development of multiple
myeloma pathogenesis
9. METODOS
CITOMETRÍA DE FLUJO
→ Fundamento: analiza una población celular según
parámetros como el tamaño celular, medidas de
epítopes (CD) o mecanismos de fluido de membrana.
→ Principio: suspensión de cells en sln isotónica →
Pequeño orificio: fila india → Se hace incidir un haz de
luz láser → Análisis de dispersión y reflexión.
10. WESTERN BLOT
→ Fundamento: detecta proteinas específicas en una
muestra.
→ Principio: proteínas separadas por electroforesis en
gel según tamaño (SDS-PAGE poliacrilámida gel) →
Filtro: incubación con anticuerpos → Identificación de
rxn.
11. MICROSCOPIA CONFOCAL
→ Fundamento: a través de la microscopía de
fluorescencia permite obtener imágenes
tridimensionales de células y tejidos complejos, se
apoya en la computación y los métodos electrónicos.
→ Principio: cortes ópticos finos de la muestra →
eliminación de la luz → reconstrucción tridimensional.
12. MTS ensayo
→ Fundamento: Ensayo de citotoxicidad colorimétrico
basado en la reduccion de Bromuro de 3-(4,5-
dimetiltiazol-2-ilo)-2,5-difeniltetrazol (MTT)
permitiendo medir la supervivencia y la proliferación
celular ademas de la citotoxicidad de potenciales
agentes terapéuticos.
→ Principio: Proceso realizado por la enzima
mitocondrial succinato deshidrogenasa que transforma
un compuesto amarillo (MTT/hidrofilico) en uno azul
(formazan/hidrofobico)
13. RT- PCR
→Fundamento:
Generación de gran
cantidad de DNA
(amplificación) por el
uso de una
transcriptasa inversa.
→Principio: Uso de una
hebra de RNA junto a
la transcriptasa inversa
para formar DNA .
17. Author Discussion Agreement
Kawano Y, Moschetta M,
Manier S, Glavey S, Görgün
GT, Roccaro AM, Anderson KC,
Ghobrial IM, Neri P, Ren L,
Azab AK, Brentnall M, Gratton
K, Klimowicz AC, Lin C,
Duggan P, Tassone P, Mansoor
A, Stewart DA, Boise LH,
Ghobrial IM, et al
MM is characterized by the
development of plasma tumor
cells that dynamically and
bidirectionally interact with their
bone marrow microenvironment.
The genetic and phenotypic
heterogeneity of MM may
consequently operate at the level
of tumor cell/tumor
microenvironment interactions.
✔
Neumeister P, Pixley FJ, Xiong
Y, Xie H, Wu K, Ashton A,
Cammer M, Chan A, Symons M,
Stanley ER, Pestell RG. Zhong
Z, Yeow WS, Zou C, Wassell R,
Wang C, Pestell RG, Quong JN,
Quong AAMeng H, Tian L, Zhou
J, Li Z, Jiao X, Li WW,
Plomann M, Lisanti MP, Wang
C, Pestell RG
Several studies have reported
that cyclin D1 controls cell
adhesion and migration of various
cell types and tumors and,
consequently, their metastatic
potential.
✔
18. Author Discussion Agreement
Goel A, Spitz DR, weiner
GJ. Manipulation of
cellular
redox parameters for
improving therapeutic
responses in
B-cell lymphoma and
multiple myeloma. J Cell
Biochem.
2012, 113:419–425
Myeloma cell death can be achieved
through the generation of ROS that
follows ER stress and UPR activation.
X
Cahu J, Bustany S, Sola B.
Senescence-associated
secretory
phenotype favors the
emergence of cancer
stem-like cells.
Cell Death Disease. 2012;
3:e446.
The common response of MM
cells exposed to X-irradiation, DNA
damaging agents, or
constitutive cyclin D1 expression is
ROS generation.
✓
19. CONCLUSION
1. Although the study only analyzed the action
of the Cyclin D1 in the parental line with
multiple myeloma, it is known that the
protein also took importance in other
pathogenesis with other response of the
expression itself, it required further
studies.
2. Cyclin D1 is an important tool because it
increases the sensibility to acute
treatments for multiple myeloma.
20. CONCLUSION
3. The redox state resulting from the
presence of cyclin D1 is necessary for the
EKR ½ pathway.
4. The Cyclin D1 has a close relationship
with the inflammatory response by the
production of cytokines such as IL8,
increases cell adhesion and promotes cell
migration.
21.
22.
23. BIBLIOGRAFÍA
• Cyclin D1 unbalances the redox status controlling cell adhesion, migration,
and drug resistance in myeloma cells. Sophie Bustany, Jérôme Bourgeais,
Guergana Tchakarska, Simon Body, Olivier Hérault, Fabrice Gouilleux,
Brigitte Sola.(07 de June de 2016). Oncotarget.-Obtenido de
http://www.impactjournals.com/oncotarget/index.php?journal=oncotarge
t&page=article&op=view&path[]=9901&pubmed-linkout=1
• Redox Status. Pedro Tauler Riera (05 February 2016) Obtenido de:
http://link.springer.com/referenceworkentry/10.1007%2F978-3-540-
29807-6_167
• LA MICROSCOPÍA: HERRAMIENTA PARA ESTUDIAR CÉLULAS Y TEJIDOS.
Obtenido de:
http://www.medic.ula.ve/histologia/anexos/microscopweb/MONOWEB/ini
cio.htm.
24. BIBLIOGRAFÍA
• PRUEBAS DE TAMIZAJE PARA DETERMINAR EFECTOS CITOTÓXICOS EN
EXTRACTOS, FRACCIONES Ó SUSTANCIAS, UTILIZANDO LA PRUEBA DEL
MTT. Clemencia Castro de Prado. Diembre 1 de 2006. Obtenido de:
http://old.iupac.org/publications/cd/medicinal_chemistry/Practica-
IV-2.pdf
• The Basics: RT-PCR. Subbu Dharmaraj, MS. 2016. Obtenido de:
https://www.thermofisher.com/co/en/home/references/ambion-
tech-support/rtpcr-analysis/general-articles/rt--pcr-the-basics.html.
• Cell-Cell Adhesion and Communication. Molecular Cell Biology. 4th
edition. 2000. W. H. Freeman and Company. Obtenido de:
http://www.ncbi.nlm.nih.gov/books/NBK21599/.
• Cell Biology of Embryonic Migration. Satoshi Kurosaka and Anna
Kashina. 2009 Jun 1. Obtenido de:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2542983/
