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Transcript of Antioxidant
that is the question! Drug Discovery From Natural Products Main topic: Breast Cancer for using Cell line Cancer Cells are
Difficult to Target for
They are very SIMILAR to normal cells in many aspects
but NOT Impossible FOR
They are also DIFFERNT from normal cells in many other ways THE MORE DIFFERENCES ARE DISCOVERED
THE MORE IS THE SURVIVING RATE At first sight,
finding a specific chemical in a "PLANT" cell to target a specific pathway or molecule particularly in a malicious "ANIMAL" cell, looks far improbable !
But on the other hand:
Thousands of plant species containing thousands of primary and secondary chemical beside thousands of pathways have altered in cancer cells has made this approach practical!! Jering (seed and pericarp) Putat Curry Borage Organic Solvents
Water 5 x 4 = 20 extracts Antioxidant Assays
Secondary Compounds Evaluation
TFC MTT assay
Active extracts are identified
IC50 is calculated HPLC & LC-MS
Identification Pathway probable suggestion Identification of
anticancer activity gene expression Protein expression Enzyme activity Active Compound breakdown Improve our underestang of Medicinal plants
in terms of
Their influence on gene/protein expression
Which would be useful for further studies •A. Kumar , R. Ilavarasan , T. Jayachandran , M. Decaraman , P. Aravindhan , N. Padmanabhan and M.R.V. Krishnan, “Phytochemicals Investigation on a Tropical Plant, Syzygium cumini from Kattuppalayam, Erode District, Tamil Nadu, South India”, Pakistan Journal of Nutrition 8 (1): 83-85, 2009.
•American cancer society, “breast cancer”, Last Revised, www.cancer.org, 2009.
•Bess L Chau and Kevin AW Lee, “Function and anatomy of plant siRNA pools derived from hairpin transgenes”, Plant Methods, 3:13, 2007.
•Coldren,-C-D; Hashim,-P; Ali,-J-M; Oh,-S-K; Sinskey,-A-J; Rha,”Gene expression changes in the human fibroblast induced by Centella asiatica triterpenoids", C, 2003.
•Deepak A.V. and Bharathi P. Salimath,” Antiangiogenic and proapoptotic activity of a novel glycoprotein from U. indica is mediated by NF-kB and Caspase activated DNase in ascites tumor model”, Biochimie. Volume 88, Issues 3-4, Pages 297-307
•Hirano T, Fukuoka K, Oka K, Naito T, Hosaka K, Mitsuhashi H, Matsumoto Y., “Antiproliferative activity of mammalian lignan derivatives against the human breast carcinoma cell line, ZR-75-1”. Cancer Invest. 8, pp. 592–602, 1990. Acknowledgments compare to pathogenes Research Title: The Investigation of the effects of selected plant extracts on the gene expression profile and the proliferation of normal & cancer cell lines
Maysam Hafezparast Mavaddat model for
bioactivity directed assays Method:
Plants are dried,
method Antioxidants Benefits
Protects cell from probable damage caused by free radicals, e.g. DNA damage
Promote faster recovery of disease
Prevent premature aging
Improve immune system function
Many exhibit anticancer properties For Better Understanding of Our Medicinal Plants Objective #1: investgation of the Antioxidant Activity of Obtained Extracts Bioactivity Test & Microscopical Observations Objective #11: Investigation of Bioactivity
of the plant extracts on normal and cancer cells bioactivity?
Anti-Cancer effect Objective #111: Isolation and Identification of active compound/compounds if any! Antioxidant Assays Proliferation Assays Few Extracts (eg. 4)
for Further Assays will be selected Bioassay Guided (eg. MTT) Objective #1V eg. Microarray, real-time PCR e.g. 2-D gel protein profiling, Western blotting, ELISA e.g. Caspase family activity e.g. using recombinant enzymes etc, Big Six of becoming a cancer cell
1- Self-sufficiency in proliferative growth signals
2- Insensitivity to growth inhibitory signals
3- Evasion of apoptosis
4-Acquisition of limitless replicative potential
5-Induction of angiogenesis
6-Induction of invasion and Metastasis Cancer is a leading cause of DEATH One of the reasons that cancer researches are so prized is References References: • Hsu, S. H., Tsai, T. R., Lin, C. N., Yen, M. H. and Kuo, K. W., ”Solamargine Purified fromSolanum incanumChinese Herb Triggers Gene Expression of Human TNFR I Which May Lead to Cell Apoptosis”, Biochem. Biophys. Res. Commun., 229, 1–5, 1996.
• Inone, I., Suzuki, R., Koide, T., Sakaguchi, N., Ogihara, Y. and Yabu, Y., “Antioxidant, gallic acid, induces apoptosis in HL-60RG cells”, Biochem. Biophys. Res. Commun., 204, 898–904. 1994.
• Jachak S. M. and A. Saklani, “Challenges and opportunities in drug discovery from plants”, CURRENT SCIENCE, VOL. 92, NO. 9, 10 MAY 2007.
• Kuo, Y. C., Sun, C. M., Tsai, W. J., Ou, J. C., Chen, W. P. and Lin, C. Y., ”Chinese herbs as modulators of human mesangial cell proliferation: preliminary studies” J. Lab. Clin. Med., 132, 76–85, 1998.
• Lee, I., Nishikawa, A., Furukawa, F., Kasahara, K. and Kim, S.,”Effects of Selaginella tamariscina on in vitro tumor cell growth, p53 expression, G1 arrest and in vivo gastric cell proliferation”, Cancer Lett., Vol. 144, No. 1, 93–99, 1999.
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•Robert Mendelsohn and Michael J. Balick, “The value of undiscovered pharmaceuticals in tropical forests”, 1995.
•Shiu L. Y. , C. H. Liang2, Y. S. Huang, H. M. Sheu and K. W. Kuo, ”Downregulation of HER2/neu receptor by solamargine enhances anticancer drug-mediated cytotoxicity in breast cancer cells with high-expressing HER2/neu”, Cell Biology and Toxicology, Volume 24, Number 1 / January, 2008.
•Taraphdar A. K., Madhumita Roy and R. K. Bhattacharya,” Natural products as inducers of apoptosis: Implication for cancer therapy and prevention”, CURRENT SCIENCE, VOL. 80, NO. 11, 10 JUNE 2001.
•Thliamplam, UM, UR Kuppusamy, M.S.Kanthimathi and C. Wiart , "Comparison between the effects of Syzygium Jambolanum leaf and fruit extracts on estrogen positive and estrogen negative breast cancer cell lines", Int. J of Molecular
Medicine and Advance Sciences, Vol. 2, no. 2 : 23. 2006.
•Tie-gang Liu, James Q Yin, Bo-yang Shang, Zhang Min, Hong-wei He, Jian-ming Jiang, Fang Chen, Yong-su Zhen, and Rong-guang Shao, “Silencing of hdm2 oncogene by siRNA inhibits p53-dependent human breast cancer”, 2004.
•Vasilis Androutsopoulos, Randolph RJ Arroo, John F Hall, Somchaiya Surichan and Gerry A Potter, “Antiproliferative and cytostatic effects of the natural product eupatorin on MDA-MB-468 human breast cancer cells due to CYP1-mediated metabolism”, Available online http://breast-cancer-research.com/content/10/3/R39, 2007.
• References: General Goals of the Study (3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide) 2,2-Diphenyl-1-picryldydrazyl free radical Ferric Reducing Antioxidant Power 2,2’-Azinobis (3-ethylbenzothiozoline-6-sulphonic acid) Total Phenolic Compound Total Flavonoid Comound What we mean when we say normal cells instruction in facing various chemicals CHANCE
HOPE between normal & cancer cell biology Supervisors: Dr. Johari Bin Mohd Ali and Dr. Azlina Binti Abdul Aziz