A Research Work on Apoptosis and Survivin
This article is devised to have an insight about the role of survivin, a protein of inhibitor of apoptosis family, in fighting the cancer. Apoptosis is a type of cell death in which the cell uses specialized cellular machinery to kill itself; a cell suicide mechanism that enables metazoans to control cell number and eliminate cells that threaten the animal’s survival. Survivin is known for its two functions- suppression of apoptosis and regulation of cell division. Recent studies have shown that over-expressed survivin has exhibited a protective action against the drug-induced apoptosis, which in many cases, lead to breast and other types of cancer. The results have further demonstrated that if the movement of survivin is restricted only to nucleus of cells, the impact of chemotherapeutic drug on a cancer patient is most effective (S. Fukuda and L. M. Pelus 2006).
Four methods are used for this study – Tissue Culture is used to generate the cancer cells in laboratory as the cancer cells have the tendency to divide the other normal cells resulting in the formation of tumor (ehow 2012). Substances used were MDA-MB-231, MDA-MB-468, BT20, and 293T cells were cultured in Dulbecco’s Modified Eagles Medium (DMEM) supplemented with 10% v/v Foetal Bovine Serum (Invitrogen), 1% v/v Penicillin/Streptomycin, 1% v/v glutamine.
Sub-cellular Fractionation enables protein localization assessment and protein enrichment from specific cellular compartments. Contents used for this are 1 ml of hypotonic buffer;10 mM Tris–Cl pH 7.5, 10 mM NaCl, 1.5 mM MgCl2, 1:100 PIC (Protease Inhibitor Cocktail, Calbiochem).
Immuno-blotting is used to detect specific proteins in the given sample of tissue homogenate or extract and Primary antibodies used for protein detection were: 1/1000 rabbit anti-Apaf-1 (Stressgen Bioreagents); 1/1000 rabbit anti-Histone H3 and 1/1000 mouse anti-Survivin (Cell Signalling Technology); 1/1000 mouse anti-HSP 70 (Affinity Bioreagents); 1/150 rabbit anti-X-IAP (Cell Signalling Technology).
Over-expression of surviving employs substances such as QIAGEN QIA prep-purified pc DNA 3.1 vectors (Invitrogen) encoding Survivin-GFP, Survivin-L98A-GFP or GFP alone, using Lipofectamine (Bioscience Horizons 2008; ehow 2012).
Over the course of past three decades, the study of apoptosis (programmed cell death) has gained utmost importance in biologists and medical sciences’ sphere. Feared by many doctors that unregulated apoptosis could lead to several diseases including cancer, neurodegenerative diseases, autoimmune disorders, and AIDS, the search for remedies combating the ill-effects of unregulated apoptosis soared exceptionally as many researchers started working on these cases. In order to devise therapeutic means to intervene and reset the balance between cell survival and death, it was important for many scientists to study the cases of unregulated apoptosis thoroughly.
As far as the beneficiaries of these studies are concerned, we can say that oncology could be the most important of them all because a lot of cancer cells show signs of suicidal machinery, also known as deregulated apoptosis. A very large beneficiary of apoptosis research is oncology, since most cancer cells exhibit defects in their suicidal machinery (EC Lacasse, S Baird, RG Korneluk et al.1998). If researchers take serious interest in deregulated apoptosis, new ways could be developed to kill the cancerous cells in the body (QL Deveraux, N Roy, HR Stennicke et al. 1998). On the other hand, pharmacologic interference with the induction or completion of apoptosis holds promise for the treatment of several neurodegenerative disorders.
The main discovery of this research work by Helen Angell is that over-expressed Survivin can protect breast cancer cells against apoptosis induced by some chemotherapeutic drugs, but only if Survivin is present within the cytosol.
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