How to Treat Breast Cancer With Turmeric - Research Proposal Example

In spite of reaching the very zenith of both technological and medical research, humans have failed to solve health problems such as cancer which continues to claim lives all around the world. Instances of breast cancer is prevalent among women and rising rapidly. However, epidemiological studies have shown that in Countries such as India and other south East Asian countries , the rates of breast cancer occurrence is far below the rates of their counterparts in the Western World. Thorough studies had been conducted to understand the exact reason for this discrepancy. The reason has been traced to nutritional intake. In spite of the nutritional intake of the Oriental world being far lower than the Western world in terms of calories and other health supplements, the presence of spices such as turmeric and other rhizomes have a drastic effect on the health. Turmeric, has long been used as a medicine in traditional Ayurveda and is today considered a potential candidate for treating cancer naturally. In this experiment, cucurmin, the active chemical derived from turmeric and known to have inhibitory effect on cancer cells, will be used to treat breast cancer cell lines in different concentrations. The results will be recorded and analyzed to understand the positive effects of in taking turmeric on a daily basis and the most effective concentration of cucurmin. CONTENTS Topic Page Introduction 3 Samples and Methods 5 Assay 5 Timeline 6 Outcomes and Implication 7 References Appendix Introduction Breast cancer is one of the leading forms of cancer responsible for high rates of mortality and morbidity among the patients especially women (CDC, 2008). Breast cancer is the most common forms of cancers responsible for deaths among women (Key et al, 2011). In 2012, in the U.K. alone almost 11,643 females and 73 males died from invasive breast cancer. Breast cancer is defines by the uncontrolled growth of the cells present in the breast area. In some cases they enter the blood stream and become invasive by affecting other organs as well. Being diagnosed with breast cancer not only reduces a person’s life span but also diminishes the quality of life that a person leads. Today, medical science has advancements such as surgery, radiotherapy, chemotherapy, hormonal therapy and gene therapy to help fight against cancer but each one of these has severe side effects (Sinha et al, 2012). Ayurveda, the study of using plants for treatment is today regarded as one of the oldest and concrete forms of treatment. In traditional India Ayurveda, people have used Cucurma longa or turmeric to treat inflammations. Besides being used for treating diseases and infections, the gold colored spice is also employed for food preservation and as a natural dye for textile (Aggarwal et al, 2007). The root of the turmeric plant is sued as a spice in many Asian cooking. The efficacy of using turmeric to treat tumorous growth has been research extensively all of which have yielded positive results. Turmeric primarily consists of three different cucurminoids- curcumin demethoxycurcumin, and bisdemethoxycurcumin out of which cucurmin is known to have inhibitory effects on cancer (Jurenka,2009). The chemotherapeutic effect of cucurmin which is a derivative of turmeric and responsible for turmeric’s yellow color, has been reported extensively (Wright et al, 2013). Cucurmin is chemically called 1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione and is known to have antioxidant, antiproliferative and apoptotic properties (Liu and Chen, 2013). Studies on rodents showed that cucurmin alone or in combination in with other chemotherapeutic agents could inhibit tumor growth (Kunnumakkara et al, 2008). Rat model experiments have shown that the rhizome of the spice inhibited cell growth in Chinese hamster Ovary cells and was cytotoxic to lymphocytes as well as Dalton’slymphoma cells at a concentration of 4 µg/ml (Kuttan et al, 1985). Therefore it was concluded that turmeric has the potential to prevent tumor growth in animal models. In another study by Banerjee et al, it was recorded that cucurmin arrested the cell division at G2/M and induced apoptosis. It also inhibited proliferation of cells and activated mitotic checkpoints (Banerjee et al, 2010). Specific breast cancer cell line studies have also shown that cucurmin had an anti-cancer activity. Hassan and Daghestani conducted an investigated to see the effect cucurmin had on the matric metalloproteinases (MMPs) and its regulator, tissue inhibitor metalloproteinases (TIMPs). The results showed that cucurmin inhibited cancer by down regulating TIMP1 and TIMP4 and up regulating MMP-2 and MMP-9 (Hassan and Daghestani,2012). It was not only animal models which responded positively, in-vitro cell line experiments using human cell line cultures have also been studied thoroughly. Some studies have shown turmeric could affect cancer growth of cells by regulating genes. In an investigation sing MCF-10A and MCF-7 it was seen that cucurmin could upregulated 22 genes (HIAP1, CRAF1, TRAF6, CASP1, CASP2, CASP3, CASP4, HPRT, GADD45, MCL-1, NIP1, BCL2L2, TRAP3, GSTP1, DAXX, PIG11, UBC, PIG3, PCNA, CDC10, JNK1 and RBP2.) and downregulated 17 genes (TRAIL, TNFR, AP13, IGFBP3, SARP3, PKB, IGFBP, CASP7, CASP9, TNFSF6, TRICK2A, CAS, TRAIL-R2, RATS1, hTRIP, TNFb and TNFRSF5). The regulation of the genes caused apoptosis in the cancer cells thereby making cucurmin a potential chemopreventive agent (Ramachandran et al, 2005). Various other studies have shown cucurmin acting by regulating genes in order to inhibit cancer. Chen et al, conducted an investigation to determine the molecular targets of cucurmin on lung cancer and established that cucurmin had the potential to inhibit invasion of cancer and prohibit migration of the cells i.e. the process of metastasis making cucurmin a potential candidate for metastatic cancer therapy (Chen et al, 2008). The primary aim of the proposed experimental study is to understand if turmeric has an effect on breast cancer cells. The study also aims to analyze and determine the lowest concentration at which the active substance of turmeric is most effective and induced apoptosis in the tumour cells Samples and Methods. The rationale for the experiment is conducting an in vitro investigation using human breast carcinoma cell lines. The cell lines that shall be employed for the experimental studies are the MCF-7 carcinoma cell lines. The active ingredient that according to research studies have anti-cancer effects on the carcinoma cells is cucurmin and hence cucurmin extracts will be used for the investigation under laboratory conditions. In order to investigate both the cytotoxic and apoptotic effects of cucrmin extract on the selected breast carcinoma cell line. The prime objective of the study is to show the inhibitory effect of turmeric on breast cancer cells in a dose dependent and time dependent manner.The cells will be cultured separately. One of the dishes will serve as the Control, to ensure proper growth of the culture and the others will the experimental dish cultures. Cucurmin will be extracted from turmeric or purchased. After the culture shave been successfully established. 7 dishes will be seeded with 500 viable cells each from the breast cancer cell line.1 of the dishes serves as the control while the others are experimental. The dishes will be incubated at different concentrations of cucurmin ranging from 10µM to 60 µM. for about 12-24 hours. The cells will then be prepared for analysis. Assay Different assays will be conducted to analyse the results. MTT assay will be conducted to obtain the growth inhibition ratio while using flow cytometry we will see the apoptotic activity. Timeline: The time and duration of each of the task of the project is displayed in the Gantt chart below. Outcomes / implications: Thorough research of the possible positive effect of turmeric on cancer has yielded good results. Considering that the efficacy of turmeric in inhibiting cancer is high, it is expected that the experimental approach designed and adopted will yield positive results as well. It is further expected that apoptotic activity will be recorded in the breast cancer cells and the concentration at which the chemical supplement is most effective can be obtained. The success of this experiment will be enough to provide sound evidence of the positive effects of using oriental spices such as turmeric in the western cuisine as well, since turmeric will work naturally without causing negative health effects. This may initiate women in U.K, to either ropt for turmeric supplements or introduce usage of turmeric in their daily meals. References: Aggarwal,B.B.,Sundaram,C., Malani,N., and ichikawa,H. 2007. Curcumin: the Indian solid gold. Advances in Experimental medicine and Biology, 595,pp.1-75. Centers for Disease Control and Prevention. 1999–2008 Incidence and Mortality Web-Based Report.Atlanta GA: Department of Health and Human Services, Center for Disease Control and Prevention, and National Cancer Institute; [Accessed 5 April,2015]. Available at:http://www.cdc.gov/uscs. Chen, H.W, Lee, J.Y, Huang, J.Y, Wang, C.C, Chen, W.J, Su, SF., Huang, C.W., Ho. C.C., Chen. J.J., Tsai, M,F., Yu, S.L., and  Yang PC. 2008. Curcumin inhibits lung cancer cell invasion and metastasis through the tumor suppressor HLJ1. Cancer Research, 68(18), pp.7428-7438. Hassan,Z.K. and Daghestani,M.H. 2012. Curcumin effect on MMPs and TIMPs genes in a breast cancer cell line. Asian pacific Journal of Cancer Prevention, 13(8),pp.3259-3264. Jurenka,J.S.2009. Anti-inflammatory properties of curcumin, a major constituent of Curcuma longa: a review of preclinical and clinical research. Alternative medicine review, 14 (2),pp.141-153. Key,T.J., Verkasalo,P.K. and Banks, E.2001. Epidemiology of breast cancer.The Lancet oncology, 2 (3),pp.133-140.  Kunnumakkara,A.B., Anand. P., and Aggarwal, B.B. 2008. Curcumin inhibits proliferation, invasion, angiogenesis and metastasis of different cancers through interaction with cell signaling proteins. Cancer Lett.269,pp199–225. Kuttan,R.,Bhanumathy,P.,Nirmala,K. and George, M.C., 1985. Potential anticancer activity of turmeric (Curcuma longa).Cancer letters, 29 (2),pp.197-202. Liu,D and Chen,Z.2013. The Effect of Curcumin on Breast Cancer Cells. Journal of Breast cancer, 16(20),pp.133-137. Ramachandran,C., Rodriguez,S., Ramachandran.R.,Raveendran Nair,P.K.,  Fonseca H, Khatib Z, Escalon E, Melnick SJ. Expression profiles of apoptotic genes induced by curcumin in human breast cancer and mammary epithelial cell lines. Anticancer Research, 25 (5),pp.3293-3302. Sinha,D, Biswas. J, Sung.B, Aggarwal, B.B and Bishaye, A.2012. Chemopreventive and chemotherapeutic potential of curcumin in breast cancer. Current Drug Targets, 13 (14),pp.1799-1819. Laura E. Wright, Jen B. Frye, Bhavana Gorti, Barbara N. Timmermann, and Janet L. Funk. . 2013. Bioactivity of Turmeric-Derived Curcuminoids and Related Metabolites in Breast Cancer. Current Pharm Des,19 (34),pp8218-6225. Appendix RESEARCH ETHICS APPLICATION FORM (STAGE 1) More information on ethics procedures can be found on your faculty website. You must read the Question Specific Advice for Stage 1 Research Ethics Approval form. All research carried out by students and staff at Anglia Ruskin University and all students at our Franchise Associate Colleges, must comply with Anglia Ruskin University Policy and Code of Practice for the Conduct of Research (students at other types of Associate College need to check requirements). There is no distinction between undergraduate, taught masters, research degree students and staff research. All research projects, including pilot studies, must receive research ethical approval prior to approaching participants and/or commencing data collection. Completion of this Stage 1 Research Ethics Application Form is mandatory for all research applications*. It should be completed by the Principal Investigator in consultation with any co-researchers on the project, or the student in consultation with his/her research project supervisor. *For research only involving animals please complete the Animal Ethics Review Checklist instead of this form. All researchers should: Ensure they comply with any laws and associated Codes of Practice that may be applicable to their area of research. Ensure their study meets with relevant Professional Codes of Conduct. Complete the relevant compulsory research ethics training. Refer to the Question Specific Advice for the Stage 1 Research Ethics Approval. Consult the Guidelines for Applying for Research Ethics Approval at Anglia Ruskin University. If you are still uncertain about the answer to any question please speak to your Dissertation Supervisor/Supervisor, Faculty Research Ethics Panel (FREP) Chair or the Departmental Research Ethics Panel (DREP) Chair. Section 1: RESEARCHER AND PROJECT DETAILS Researcher details: Name(s): Department: Faculty: Anglia Ruskin email address: Status: Undergraduate Taught Postgraduate Postgraduate Research Staff If this is a student project: SID: Course title: Supervisor/tutor name Project details: Project title (not module title): Data collection start date: (note must be prospective) Expected project completion date: Is the project externally funded? Licence number (if applicable): CONFIRMATION STATEMENTS – please tick the box to confirm you understand these requirements The project has a direct benefit to society and/or improves knowledge and understanding. All researchers involved have completed relevant training in research ethics, and consulted the Guidelines for Applying for Ethical Approval at Anglia Ruskin University. The risks participants, colleagues or the researchers may be exposed to have been considered and appropriate steps to reduce any risks identified taken (risk assessment(s) must be completed if applicable, available at: http://rm.anglia.ac.uk/extlogin.asp) or the equivalent for Associate Colleges. My research will comply with the Data Protection Act (1998) and/or data protection laws of the country I am carrying the research out in, as applicable. For further advice please refer to the Question Specific Advice for the Stage 1 Research Ethics Approval. Project summary (maximum 500 words): Please outline rationale for the research, the project aim, the research questions, research procedure and details of the participant population and how they will be recruited. The research experiment aims to find out the efficacy of cucurmin extracts in treating breast cancer cells and the effective concentration at which apoptotic activity is induced. The research questions to be answered are: 1. Is cucurmin effective against breast cancer? 2. If it is effective then at what concentration? Experimental procedure: Breast cancer cell line will be cultured under laboratory settings. Cucurmin shall be purchased and different concentrations of cucurmin will be prepared. The established cell culture will be divided into 7 dishes with each dish containing 500 viable cells. Each dish will be treated with cucurmin extract. The control will not be treated with any cucurmin extract. After 24 hours the cells will be prepared for analysis. The analysis will be done suing flow cytometer and MTT assay. Sample: The breast cancer cell line will be purchased. Section 2: RESEARCH ETHICS CHECKLIST - please answer YES or NO to ALL of the questions below. WILL YOUR RESEARCH STUDY? YES NO 1 Involve any external organisation for which separate research ethics clearance is required (e.g. NHS, Social Services, Ministry of Justice)? 2 Involve individuals aged 16 years of age and over who lack capacity to consent and will therefore fall under the Mental Capacity Act (2005)? 3 Collect, use or store any human tissue/DNA including but not limited to serum, plasma, organs, saliva, urine, hairs and nails? Contact matt.bristow@anglia.ac.uk 4 Involve medical research with humans, including clinical trials? 5 Administer drugs, placebos or other substances (e.g. food substances, vitamins) to human participants? 6 Cause (or could cause) pain, physical or psychological harm or negative consequences to human participants? 7 Involve the researchers and/or participants in the potential disclosure of any information relating to illegal activities; or observation/handling/storage of material which may be illegal? 8 With respect to human participants or stakeholders, involve any deliberate deception, covert data collection or data collection without informed consent? 9 Involve interventions with children and young people under 16 years of age? 10 Relate to military sites, personnel, equipment, or the defence industry? 11 Risk damage or disturbance to culturally, spiritually or historically significant artefacts or places, or human remains? 12 Involves genetic modification, or use of genetically modified organisms above that of routine class one activities? Contact FST-Biologicalsafety.GMO@anglia.ac.uk (All class one activities must be described in section 4). 13 Contain elements you (or members of your team) are not trained to conduct? 14 Potentially reveal incidental findings related to human participant health status? 15 Present a risk of compromising the anonymity or confidentiality of personal, sensitive or confidential information provided by human participants and/or organisations? 16 Involve colleagues, students, employees, business contacts or other individuals whose response may be influenced by your power or relationship with them? 17 Require the co-operation of a gatekeeper for initial access to the human participants (e.g. pupils/students, self-help groups, nursing home residents, business, charity, museum, government department, international agency)? 18 Offer financial or other incentives to human participants? 19 Take place outside of the country in which your campus is located, in full or in part? 20 Cause a negative impact on the environment (over and above that of normal daily activity)? 21 Involve direct and/or indirect contact with human participants? 22 Raise any other ethical concerns not covered in this checklist? Section 3: APPROVAL PROCESS Section 4: ETHICAL RISK (Risk category 2 projects only) Management of Ethical Risk (Q14-22) For each question 14-22 ticked ‘yes’, please outline how you will manage the ethical risk posed by your study. Section 5: Declaration *Student/Staff Declaration By sending this form from My Anglia e-mail account I confirm that I will undertake this project as detailed above. I understand that I must abide by the terms of this approval and that I may not substantially amend the project without further approval. **Supervisor Declaration By sending this form from My Anglia e-mail account I confirm that I will undertake to supervise this project as detailed above. *Students to forward completed form to their Dissertation Supervisor/Supervisor. ** Dissertation Supervisor/Supervisor to forward the completed form to the relevant ethics committee. May 2014 Version: 5.0 Read More