Академический Документы
Профессиональный Документы
Культура Документы
The cell can either repair itself or programmed itself to die, Apoptosis is one of the mechanisms that execute the cell to die.it has two
general pathways the intrinsic and extrinsic pathways.
In intrinsic pathway , the death inducing stimuli is originated inside the target cell itself.Mitochondria, the powerhouse of the cell have
significant role in executing the intrinsic pathway of apoptosis, Thus the intrinsic pathway is also known as Mitochondria-mediated
death pathway [1].The initiation begins with the internal stimuli such as severe DNA damage, lack of oxygen and presence of viral
proteins then it will be sense by some signalling proteins molecule like ATM and CHk which will activate the p53 protein which caused
to halt the cell cycle through recruiting other mediator proteins like the BAX and Bad (The pro apoptosis protein), it was the protein that
creates pores in mitochondria, once pores form in mitochondria and became permeable , cytochrome C will be release to the cytosol,
the complete release of cytochrome c to the cytosol is a point of no return , it won’t be reverse to normal cell moreover it will commit
apoptosis. cytochrome -C will be bind with APAF-1 and Pro-caspase 9 in the cytosol to form multi-subunit complex called
APOPTOSOME and binding of apaf-1 induce conformational change in the pro caspase 9 and it is activated to its fully proteolytic form
called caspase 9. Caspase 9 activates the executioner caspases such as caspase 3 and caspase 7, once it was activated it degrades
the inhibitor of the nucleus then nuclease activity proceeds which is destroying the cell such as degrading the DNA [2]. Apoptosis
executed
Extrinsic Pathway
In extrinsic pathway of apoptosis, the death inducing signal is triggered by external stimulus. The cell possesses plasma membrane
receptors to receive the external stimuli, in most cases is a cytokine specifically a TNF (TUMOR - NECROSIS FACTOR) which is
produce by immune system in response to conditions such as viral toxins, infections, radiation exposure. TNF will bind to its receptor
called TNFR-1 present on the plasma membrane it has external ligand binding domain and a cytosolic domain, the cytosolic domain
contains 70 amino acids called death domain. The binding of the external stimuli(TNF) to the receptor (TNFR1) cause a conformational
change in death domain which cause a recruitment of apoptosis adaptor proteins such as TRADD and FADD,TRADD and FADD
adaptor proteins are binds with Pro-Caspase 8 residue to form a multi-protein complex.Interactions of death effector domains present
in both Pro caspase 8 and FADD generate an active caspase 8 which activate the initiator caspase ,the caspase 3 , same way in the
intrinsic pathway , caspase 3 will carry out the apoptosis or self-destruction process.[3]
References
[1]https://www.easybiologyclass.com/intrinsic-pathway-of-apoptosis-apoptosis-molecular-mechanism-part-1/
[2] Shomus biology
[3]https://www.easybiologyclass.com/extrinsic-pathway-of-apoptosis-apoptosis-molecular-mechanism-part-2/
Increased in IFN-g Secretion via enhanced natural killer (NK) cells was caused by andrographolide and this NK- mediated in
killing the non– small cell lung cancer (NSCL) cells in membrane bound in certain cellular molecules. To discuss further how
andrographolide affects apoptosis of NSCL cells it was stated in the journal that, Apoptosis is mediated through up-regulation of Bax,
Bad, p53, Caspases-3, -9, and cleavage of poly (ADP-ribose) polymerase (PARP) and down regulation of Bcl-2, cyto-solic NF-kB p65,
phosphati-dylinositol-4,5-bisphosphate 3-kinase (PI3K), and p-AKT in many cell lines such as colorectal carcinoma (HCT116),
pancreatic carcinoma (MiaPaCa-2), HepG2, cervical carcinoma (HeLa), lung carcinoma (A549), and melanoma (A375) cells.
Furthermore, in focus on lung cancer cell lines, when andrographolide was co-treated with paclitaxel in A549 cells (5.10e328.0 mM þ
0.48e60.75 nM) and xenograft murine model (100 mg/kg þ 20 mg/kg), it showed a significant synergistic effect on ROS accumulation
and G2/M phase cell cycle arrest which cause apoptosis of the said carcinoma. On the other hand, one of the andrographolide
derivative namely, 3,19-diacetyl-14-deoxy-11,12-didehydro-Andro effectively suppressed nuclear translocation of NF-kB in A549 cells
and decreased pulmonary eosinophilia, bronchoalveolar lavage (BAL)fluid inflammatory cytokines level, serum immunoglobulin (Ig)-E
production, mucus hypersecretion and gene expression of inflammatory mediators in lung tissues in ovalbumin (OVA)-induced asthma
model in female BALB/c mice. Moreover, in NSCLC, andrographolide repressed the expression of matrix metallo-protein (MMP)-2 and
affected the expression of 939 genes leading to cell cycle arrest and apoptotic cell death.
Definition of terms
IFN-g (interferon gamma) - This gene encodes a soluble cytokine that is a member of the type II interferon class. The encoded protein is
secreted by cells of both the innate and adaptive immune systems. The active protein is a homodimer that binds to the interferon gamma
receptor which triggers a cellular response to viral and microbial infections
https://www.ncbi.nlm.nih.gov/gene/3458
NF-kb(nuclear factor kappa B subunit 1) - is a protein complex that controls transcription of DNA, cytokine production and cell survival.
NF-κB is found in almost all animal cell types and is involved in cellular responses to stimuli such as stress
https://en.wikipedia.org/wiki/NF-κB
PI3K-Akt Pathway is a signal transduction pathway that promotes survival and growth in response to extracellular signals.
https://en.wikipedia.org/wiki/Akt/PKB_signaling_pathway
reactive oxygen species (ROS)- are chemically reactive chemical species containing oxygen. Examples include peroxides, superoxide,
hydroxyl radical, singlet oxygen, and alpha-oxygen. In a biological context, ROS are formed as a natural byproduct of the normal
metabolism of oxygen and have important roles in cell signaling and homeostasis. However, during times of environmental stress, ROS
levels can increase dramatically. This may result in significant damage to cell structures.
https://en.wikipedia.org/wiki/Reactive_oxygen_species
Paclitaxel is a cyclodecane isolated from the bark of the Pacific yew tree, TAXUS BREVIFOLIA. It stabilizes MICROTUBULES in their
polymerized form leading to cell death.
https://pubchem.ncbi.nlm.nih.gov/compound/paclitaxel
Xenograft murine model-is used to test the efficacies of chemo treatments or gene therapy in an in vivo system. In this protocol, the mouse
xenograft model is used.
https://bio-protocol.org/e22
A synergistic effect is the result of two or more processes interacting together to produce an effect that is greater
https://www.safeopedia.com/definition/517
Matrix metalloproteinases (MMPs)- these enzymes are capable of degrading all kinds of extracellular matrix proteins, but also can
process a number of bioactive molecules. They are known to be involved in the cleavage of cell surface receptors, the release of
apoptotic ligands (such as the FAS ligand), and chemokine/cytokine inactivation.
https://en.wikipedia.org/wiki/Matrix_metalloproteinase
Autophagy
Auto means self, Phagy means eating , thus autophagy is a self-eating process, it is one of the major degradation process of proteins
in mammalian cell , It has 3 types, the macro autophagy that simply means degradation of bulk cellular components, micro autophagy
and chaperone – mediated autophagy ,this 3 types has common mechanism, it is the degradation of substrate within lysosome. Since it
is degradation process, it is responsible in cellular waste clearance moreover, it recycles nutrients sub unit such as amino acids,
another related process that can occur in autophagy is the autophagy cell death.
Reference
https://www.youtube.com/watch?v=UmSVKzHc5yA&t=67s( JJ Medicine Channel)
Reference
https://www.semanticscholar.org/paper/Inhibition-of-autophagy-by-andrographolide-cell-via-Mi-
Xiang/5ed9219e30f17558e19a0aa4b34eae4ceb9ec497
Cell adhesion
In NSCLC, it inhibited MMP-7 expression through the suppression of PI3K/AKT/AP-1 signaling pathway
leading to reduced cell invasiveness However, in NSCLC cells, the anti-invasive and anti-proliferative effects of andrographolide were
linked to its inhibitory effects on the expression of MMP-2 and other 939 genes]. In another study, andrographolide (1, 5 and
10 mM) was found to inhibit cell proliferation, along with down-regulation of the MMP-9 mRNA expression and inhibition of nu-
clear translocation of NF-kB p65 subunit through the suppression of IkB phosphorylation in H3255 lung cancer cells.Similarly,
andrographolide inhibited the migration and invasion of NSCLC A549 cells via down-regulation of PI3K/Akt signaling pathway. In
addition, it selectively exerted an inhibitory effect on the activity MMP-7 genes but not on MMP-2 or MMP-9.
Andrographolide-induced events that affects cell migration and invasion, Moreover, andrographolide in live cancer cells inhibited
metastasis through its action on NF-kB and actin proteins.