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Project Proposal

09/05
by

Monica B

on 22 May 2013

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Transcript of Project Proposal

Monica Bodd, Audrey Chang, Karen Mac (MAK) AuNP + Laser Therapy Design gold nanorod conjugates using EGFR and VEGFR antibodies to attach to cell surfaces.
Use a laser to test the effectiveness of the gold nanorods to destroy cancer cells. QUESTION/PURPOSE If lung cancer/breast cancer cells are incubated with conjugated gold nanorods and exposed to varying laser intensities at 808nm, then at least twice the amount of lung cancer/breast cancer cells would die in comparison to lung cancer/breast cancer cells not exposed to laser treatment.
If both lung cancer/breast cancer and healthy lung/healthy breast cell lines are incubated with anti-EGFR/anti-VEGFR conjugated gold nanorods and exposed to varying laser intensities at 808nm, then at least twice the amount of lung cancer/breast cancer cells would die in comparison to healthy lung/healthy breast cells. HYPOTHESIS BACKGROUND &
LITERATURE REVIEW Culture Cells

PEGylate gold nanorods

Conjugate gold nanorods with EGFR and VEGFR antibodies

Use UV-Vis spectrophotometer to ensure binding of antibodies to nanorods

Immerse cells with gold nanorod solution

Use microscope with dark field condenser to gather light scattering images of AuNPs binding to cells

Count Cells

Using a cw Ti:sapphire laser at 808 nm, expose cells to laser beam for 4 minutes, at varying intensities

Dye the laser-treated cells with trypan blue

Count cells METHODS Bionanotechnology: Manipulation of Photothermal Properties of Gold Nanorod Conjugates of EGFR and VEGFR in Mammary Carcinoma and Squamous Cell Lung Carcinoma Cancer + Therapy
Stages I, II, III, IV
radiation, chemotherapy, surgery
Gold Nanoparticles
Epidermal Growth Factor Receptor (EGFR)
photothermal therapy - in vitro
Vascular Endothelial Growth Factor Receptor (VEGFR)
inhibition of growth pathway
Antibody-GNR Conjugation, Pegylation MATERIALS healthy mammary gland cells
healthy lung cells
mammary gland carcinoma
lung carcinoma
all materials necessary for gold nanorod conjugation to anti-EGFR and anti-VEGFR antibodies
UV-VIS spectrophotometer
microscope with a dark field condenser
cw Ti:sapphire laser
trypan blue stain CONSTANTS 5% CO2
37 degrees Celsius
Aspect ratio of 4.1 of Au Nanorods
Wavelength of laser at 808 nm Bibliography "Chemotherapy and Cancer Treatment Side Effects." MedicineNet. MedicineNet, Inc., n.d. Web. 19 Sept. 2012. <http://www.medicinenet.com/script/main/art.asp?articlekey=21716>.
Green, Hayidah N. et al. “Gold Nanorod Bioconjugates for Active Tumor Targeting and Photothermal Therapy,” Journal of Nanotechnology, vol. 2011, Article ID 631753, 7 pages, 2011. doi:10.1155/2011/631753
El-Sayed, Ivan H., Xiaohua Huang, and Mostafa A. El-Sayed. "Selective Laser Photo-thermal Therapy of Epithelial Carcinoma Using Anti-EGFR Antibody Conjugated Gold Nanoparticles." Cancer Letters 129-135 239 (2005): n. pag. Web. 11 Sept. 2012.
Huang, Xiaohua, Ivan H. El-Sayed, Wei Qian, and Mostafa A. El-Sayed. "Cancer Cell Imaging and Photothermal Therapy in the Near-Infrared Region by Using Gold Nanorods." JACS , 2115-2120 128 (2006): n. pag. Web. 11 Sept. 2012. <http://nanoseedz.com/references/RF-Photothermal-1.pdf>.
Kalishwaralal, K., S. Sheikpranbu, S. BarathManiKanth, R. Haribalaganesh, S. Ramkumarpandian, and S. Gurunathan. "Gold Nanoparticles Inhibit Vascular Endothelial Growth Factor-induced Angiogenesis and Vascular Permeability via Src Dependent Pathway in Retinal Endothelial Cells." National Center for Biotechnology Information. U.S. National Library of Medicine, 2011. Web. 12 Sept. 2012. <http://www.ncbi.nlm.nih.gov/pubmed/21061058>.Olson, Anna-Karin, Anna Dimberg, Johan Krueger, and Lena Claesson-Welsh. "VEGF Receptor Signaling - in Control of Vascular Function." Nature 7 (2006): 359-71. Web. <http://web.mit.edu/hst527/www/readings/Lecture 6/VEGF.pdf>.
"Radiation Therapy and You." National Cancer Institute. National Cancer Institute, 20 Apr. 2007. Web. 19 Sept. 2012. <http://www.cancer.gov/cancertopics/coping/radiation-therapy-and-you/page6>.
Zhang, Weiqi. "Probing the Behaviors of Gold Nanorods in Metastatic Breast Cancer Cells Based on UV-vis-NIR Absorption Spectroscopy." PLOS ONE. Internet Systems Consortium, 22 Feb. 2012. Web. 12 Nov. 2012. <http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0031957>. EGFR Experiment & VEGFR Experiment
no treatment (control) versus with treatment
dependent
number of dead noncancerous/cancerous cells after laser therapy
independent
treatment/no treatment
cell line
antibody OVERVIEW CONCLUSION Our results support all of the hypotheses stated.
The cancerous cells that underwent treatment had a statistically significant difference in cell death compared to control samples.
Lung cancer cells incubated with anti-EGFR conjugated gold nanorods had the most statistically significant cell death difference due to laser treatment.
Laser treatment affected the cancer cell lines more than the healthy cell lines because of the presence of growth factor receptors, EGFR and VEGFR. April 13, 2013 RESULTS LC: Lung cancer with treatment
LC ANL: Lung cancer without treatment BC: Breast cancer
BC ANL: Breast cancer without treatment
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