Abstract

1. Need Abstract http://www.cdc.gov/ncidod/dvbid/westnile/mapsactivity/surv&control07Maps.htm Knowledge Base: Previous studies have shown that morphine has the capabilities to both promote and inhibit tumor growth. The key factor that establishes the fine line between the two opposite occurrences is the concentration of the morphine being administered. The objective of this study was to determine the differences of the expression of certain genes at low concentrations of morphine in both healthy and cancerous cells. Data from a DNA microarray was analyzed on SpotFire Decision Site to identify the varying expressions of key genes that are involved in nicotinic acetylcholine receptors (nAChR). After healthy white blood cells and U937 cancer cells were exposed to physiological levels of morphine, two of these genes were discovered to be up-regulated in healthy cells and down-regulated in cancer cells, a significant distinction that could lead to implications of cancer detection and possibly alternate treatment. In addition to SpotFire data analysis, a follow-up experiment was conducted to examine the effects of morphine on healthy hemocytes treated with malathion. The results of the experiment showed a significant correlation between Malathion and the control groups when comparing form factor. Further experimentation showed that malathion mimics morphine's effect on hemocytes; this was proven by blocking its effect with Naloxone and L-name. Since opiate and nicotinic signaling systems are very similar in invertebrates and human cells, our results indicate that Malathion may affect tumor development via opiate and nicotinic signaling. The impact of Malathion on environment and human health should be further studied. • Morphine signaling system • Animal /human cells can make morphine • Morphine has is on receptor. • Morphine can couple with this receptor and release nitric oxide • Low level of morphine can down regulate immune respone • How ever, high level of morphine can stimulate tumor growth Studies have shown that nicotine can effect morphine signaling in animal and human cells The mechanism for malation to kill insects is through nicotin/actocholine receptor Low Level (Endogenous) Morphine High Level of Morphine Question There was no report that malathion effect on opiate signaling system. And the mechanism that malathion can cause cancer development is not clear. ECHIBURU-CHAU et al. 2008 Nitric Oxide Mosquito Eradication Malathion http://dailymed.nlm.nih.gov/dailymed/archives/fdaDrugInfo.cfm?archiveid=9182 Healthy state, Antiviral, Antibacterial Cellular Damage, Cancer Development http://www.jyi.org/research/re.php?id=88 http://www.aedesmosquito.com/ http://thebrain.mcgill.ca/flash/a/a_06/a_06_m/a_06_m_mou/a_06_m_mou.html http://thebrain.mcgill.ca/flash/a/a_06/a_06_m/a_06_m_mou/a_06_m_mou.html Purposes: • The purpose of this experiment is to evaluate Malathion effects on opiate morphine signaling in healthy animal cells and its implication with cancer development Hypothesis: H(a): Malathion will effect the opiate signaling pathway in hemocyte cells and therefore may show a link to cancer development. H(0): Malathion will have no effect on the opiate signaling pathway in hemocyte cells and show no possible correlation to tumorr development.

2. Microarray Analysis Analyze Microarray Data Methodology: Hemocyte Extraction Search for genes for interest Divide Hemocytes into Six-Well Plates Take down data from microarray and graph to compare gene expression percent down Malathion and Naloxone Malathion and L-name Control Morphine Malathion Morphine and Naloxone Morphine and L-name Take Pictures Calculate and Analyze Form Factor Statistical Analysis (TTest) Graph 1a: This graph shows the effect of morphine on gene expression in white blood cells. The graph shows by what percent genes are down regulated. Graph 1b: This graph shows the effect of morphine on gene expression in U937 Cells (Cancer Cells). The graph shows by what percent genes are down regulated. Figure 2a: Graph shows the difference in form factor between the control morphine group and the malathion group. Figure 2b: Graph shows morphine naloxone and l-name’s effect on cell form factor as compared to the control. Figure 2c: Graph shows naloxone and l-name’s effects on malathion contamination on form factor as compared to the control.

3. Nicotinic Signaling Morphine Malathion μ3/μ4 Receptor Discussion Nitric Oxide Syntase Healthy Cells Cancer Cells Picture 2: Malathion’s effect on hemocytes after one hour of exposure time. Picture 1: Control hemocytes cells one hour after dissection Hemocytes with Malathion Exposure Normal Hemocytes Malathion’s effect on gene expression of the μ3/ μ4 receptor over long periods of time Malathion’s effect on gene expression in both white blood cells and cancer cells • This study correlates with ECHIBURU-CHAU et al. 2008 • time may have an effect on the degree of damage malathion may cause Effects on: Conclusion Malathion effects the morphine signaling pathway either by causing morphine in the cell or directly affecting the receptor. Suggests that better precautions must be taken when using malathion and other insecticides Future Studies Bibliography Cabello Gertrudis, Valenzuela Mario, Vilaxa Arnaldo, Durán Viviana, Isolde Rudolph, Hrepic Nicolas, and Gloria Calaf; A Rat Mammary Tumor Model Induced by the Organophosphorous Pesticides Parathion and Malathion, Possibly through Acetylcholinesterase Inhibition; Environmental Health Perspectives; vol 109; pgs 471-479; May 2001 Enviormental Toxins. "Cancer Caused By Environmental Toxins."; Web. 15 Sept. 2010. <http://ezinearticles.com/?Cancer-Caused-By-Environmental-Toxins&id=648200>. Ishikawa, M., Tanno, K., Kamo, A., Takayanagi, Y., Sasaki, K., Enhancement of tumor growth by morphine and its possible mechanism in mice;Biological and Pharmaceutical Bulletin, 16, 762-766. George B. Stefano, John D. Burrill, Sergei Labur, Julie Blake, Patrick Cadet; “Regulation of various genes in human leukocytes acutely exposed to morphine: expression microarray analysis”; Med Sci Monit; Published: 2005.05.05 George B. Stefano, Richard M. Kream, Kirk J. Mantione, Melinda Sheehan, Patrick Cadet, Wei Zhu, Thomas V. Bilfinger, and Tobias Esch; “Endogenous Morphine/Nitric Oxide-Coupled Regulation of Cellular Physiology and Gene Expression: Implications for Cancer Biology”; Published in June 2008. Gupta, K., Kshirsagar, S., Chang, L, Schwartz, R., Law, PY., Yee, D., Hebbel, RP., Morphine stimulates angiogenesis by activating proangiogenic and survival-promoting signaling and promotes breast tumor growth [Electronic version]. Cancer Research, 62, 4491-4498. Loguinov A, Anderson L, Crosby G, Yukhananov R (2001). "Gene expression following acute morphine administration". Physiol Genomics6 (3): 169–81. PMID 11526201 Marilie D. Gammon et. al.; “Enviormental Toxins and Breast Cancer on Long Island. Polycyclic Aromatic Hydrocarbon DNA Adducts”; August 2002; Cancer Epidemiology, Biomarkers & Prevention; Vol. 11; 677–685. Maureen R. Gwinn, Diana L. Whipkey, Lora B. Tennant, and Ainsley Weston; Differential Gene Expression in Normal Human Mammary Epithelial Cells Treated with Malathion Monitored by DNA Microarrays; Environmental Health Perspectives; vol 113; pg 1046-1051; August 2005 Matthew R. Bonner, Joseph Coble, Aaron Blair, Laura E. Beane Freeman, Jane A. Hoppin Dale P. Sandler, and Michael C. R. Alavanja; Malathion Exposure and the Incidence of Cancer in the Agricultural Health Study; American Journal of Epidemiology; Vol. 166; Pgs 1023-1034; May 17, 2007 "UNDERSTANDING ANGIOGENESIS." Angiogenesis Foundation. Web. 17 Sept. 2010. <http://www.angio.org/ua.php>. Stefano GB, Hartman A, Bilfinger TV, Magazine HI, Liu Y, Casares F, Goligorsky MS; Presence of the mu3 opiate receptor in endothelial cells. Coupling to nitric oxide production and vasodilation; Journal of Biological Chemistry; vol 270; pgs 30290-30293; December 22, 1995 Stefano George B., Kream Richard M., Mantione Kirk J., Sheehan Melinda, Cadet Patrick, Zhu Wei, Bilfinger Thomas V., and Esch Tobias; “Endogenous Morphine/Nitric Oxide-Coupled Regulation of Cellular Physiology and Gene Expression: Implications for Cancer Biology”; Semin Cancer Biol; vol 18, pgs 199–210; June 2008 Zhu W, Cadet P, Baggerman G, Mantione KJ, Stefano GB; Human white blood cells synthesize morphine: CYP2D6 modulation; J Immunol; vol 175; pgs 7357-7362; December 2005 Zhu Yu Cheng, Snodgrass Gordon L., Chen Ming Shun; Enhanced esterase gene expression and activity in a Malathion resistant strain of the tarnished plant bug, Lygus lineolaris; Insect Biochemistry and Molecular Biology; vol 34; pgs 1175–1186; May 24 2004