My Dream Experiment - Microscopy and Live Imaging - IZN Heidelberg 2016
The effect of Inhibition Factor AY on Survivin
Expression in cancer cells A.Tajeri, Y.Ben Mansour, University of Heidelberg, 2016
Abstract Survivin, a member of the inhibitor of apoptosis
(IAP) protein family that blocks cell death, is highly expressed in most cancers and is associated with a poor clinical outcome. The survivin gene polymorphisms have also been reported to influence tumor aggressiveness as well as the survival of cancer patients. This review discusses the role of IFAY (Inhibition Factor Ashkan Yassine) in blocking of survivin`s function in cancer cell. Keywords: Cancer, Survivin, Inhibition Factor
I. INTRODUCTION
HE inhibitor of apoptosis (IAP) proteins are a family of
highly conserved cell death inhibitors that have been found in yeast, invertebrates, and vertebrates1,2. Survivin, an inhibitor of apoptosis protein IAP, shown to be involved in suppressing cell death response1,2. Currently, Survivin protein expression is being used as a prognostic factor in several human cancers3. Manipulation of Survivin regulation and expression may also lead to the development of new immunotherapy and gene therapy strategies for the treatment of cancer1,2,3. The ficticious Inhibition Factor Ashkan Yassine (IFAY), imagined by Ashkan Tajeri and Yassine Ben Mansour for My Dream Experiment, is supposed to bind transcription factors, and prevent RNA polymerase from docking to the region, thereby inhibiting transcription of the gene coding for Survivin. II. GUIDELINES FOR EXPERIMENTAL METHODS 1- Experimental System: Cancer cell cultures 2- Measurement: Interaction between the Inhibition Factor AY (IFAY) and the Survivin promoter sequence. 3- Fluorescent molecule class: genetically encoded green fluorescent protein (GFP). 4- Application method: virus-mediated transfection. Lentivirus can integrate into the host cell genome to allow stable, long-term expression4. 5- Targeting: we aim to target the promoter sequence
regulating Survivin expression.
6- Color Class: single color, GFP to detect transgenic expression in vitro. GFP can be excited by 488nm light and is optimally detected at 510nm4. 7- Microscopy method: Epifluorescence widefield microscope for a qualitative analysis. 8- Microscopy setup: GFP filter set. 9- Data analysis: a qualitative analysis. We aim to perform an experiment with 2 samples of cancer cell cultures. Both cultures are infected with lentivirus containing a Survivin promoter sequence linked to a GFP coding sequence. The transfected cells, under normal conditions, are going to express Survivin coupled with GFP, so that an emission of the Survivin-GFP complex is resulting at 510nm after excitation at 488nm. The first sample had been incubated with our Inhibition Factor (IFAY). The second sample is serving as negative control, without any inhibition factors, a Survivin-GFP complex emission is to be observed. III.
FICTITIOUS RESULT
As we expect, Inhibition Factor AY should repress the
expression of Survivin, as a result, the Survivin concentration in the cytoplasm is nearly by zero and this may lead to a dramatical reduction of GFP emission. Inhibition Factor AY seems to have a primordial role in blocking the expression of Survivin, and as a result a suppression of cell Death is shut down. We can suppose that our Inhibition Factor is an effective drug to influence tumor development. Furthermore, a quantitative analysis with ELISA could be performed to detect the fluctuations in Survivin concentration in cells. IV. CONCLUSION As a conclusion, Survivin seems to be proposed as an attractive target for new anticancer interventions. Although, some molecular functions as chaperone binding, enzyme binding and microtubule binding; as well as some biological processes like regulation of cell proliferation and regulation of
My Dream Experiment - Microscopy and Live Imaging - IZN Heidelberg 2016
apoptosis should be more understood to optimize efficient anticancer therapeutic interventions. ACKNOWLEDGMENT We want to thank Anna Hertle for encouraging students to gain self-confidence and develop imagination in designing research projects. REFERENCES [1] Altieri DC, Marchisio PC: Survivin apoptosis: an interloper between cell death and cell proliferation in cancer. Lab Invest, 1999; 79: 1327-1333 [2] Schimmer AD. Inhibitor of apoptosis proteins: translating basic knowledge into clinical practice. Cancer Res. 2004;64:718390 [3] Chiou SK, Jones MK and Tarnawski AS, Survivin an anti-apoptosis protein: its biological roles and implications for cancer and beyond. Department of Medicine, Veterans Affairs Medical Center, Long Beach, CA, USA University of California, Irvine, CA, USA, 2003. [4] www.thermofisherscientific.com