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ANTIPROLIFERATIVE STUDIES OF Euphorbia hirta ON MCF-7 AND HEP-2 CELL LINES:

ANTIPROLIFERATIVE STUDIES OF Euphorbia hirta ON MCF-7 AND HEP-2 CELL LINES By N.Radha Krishna Reddy (Msc.Microbiology) under the esteemed guidance of Dr.Varaprasad Bobbarala (chief scientist)

AIM & OBJECTIVE:

AIM & OBJECTIVE Determination of hidden potential of a crude extract of Euphorbia hirta for a specific biological activity Identification of anticancer activity of crude extract of Euphorbia hirta using MTT- [3-(4, 5-Dimethylthiazol-2-yl)-2,5-Diphenyl tetrazolium bromide] Assay

INTRODUCTION:

INTRODUCTION Introduction to the natural products : Natural product is a chemical compound or a substance produced by living organism Compounds are important in the treatment of life-threatening conditions Natural products chemistry has brought forth compounds that have gone on to eventually lead to advanced cancer treatments and a wide range of health issues

ABSTRACT:

ABSTRACT Plant part used:Aerial Part A small perennial herb with milky latex in all parts of the plant Phytochemicals:alkaloids,essential oils,phenols,sterols,flavones and fatty acids

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Properties:Anthelmintic,Expectorant, Antidysenteric,Hypoglycemic Antimalarial and Anti-tumor properties Medicinal uses:Asthma,Bronchitis,Worm infestations,conjuctivities,Dysentery

CANCER:

CANCER Cancer is one of the most common diseases in the developed world: One in four deaths are due to cancer Cancer is an abnormal growth and proliferation of cells Lung cancer is the most common cancer in men Breast cancer is the most common in women

BREAST CANCER:

BREAST CANCER Breast cancer is a cancer that starts in the tissues of the breast Ductal carcinoma starts in the tubes(ducts) that move milk from the breast to the nipple Lobular carcinoma starts in the parts of the breast called lobules that produce milk

CAUSES & RISK FACTORS :

CAUSES & RISK FACTORS Heredity Child birth Obesity Radiation Age and gender Menstrual cycle Hormone replacement therapy

SYMPTOMS:

SYMPTOMS Breast lump or lump in the armpit Change in size,shape or feel of the breast or nipple Fluid coming from the nipple may be bloody,clear to yellow Swelling of one arm next to the breast with cancer Weight loss

DIAGNOSIS:

DIAGNOSIS Breast MRI Breast ultrasound Breast biopsy Mammography PET scan CT scan

Stage 1 :

Stage 1 Tumor < 2.0 cm in greatest dimension Stage II Tumor > 2.0 < 5 cm Stage III Tumor > 5 cm (T3) Stage IV The cancer has spread to other organs of the body usually the lungs, liver, bone or brain.

TREATMENT & SIDE EFFECTS:

TREATMENT & SIDE EFFECTS Radiation therapy Chemotherapy Heperthermia treatment

Euphorbia hirta or Asthma Weed:

Euphorbia hirta or Asthma Weed The plant is native to India but is a pan tropical weed. Euphorbia hirta is a medicinal,rhizomatous herb

MATERIALS:

MATERIALS MTT- [3-(4, 5-Dimethylthiazol-2-yl)-2,5-Diphenyl tetrazolium bromide] dye Trypsin-EDTA Minimal essential medium (MEM ) T-flasks (Tissue culture flasks) Make: Corning USA 15ml centrifuge tubes - TARSONS Anthos Biochrom ELISA plate reader

METHODLOGY:

METHODLOGY Plant Extraction Soxhlet Extractor: Invented in 1879 by Franz von soxhlet For the extraction of a lipid from a solid material

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Stirrer bar Condensor Siphon top Siphon exit Condenser Distillation

SUB CULTURING PROCEDURE FOR CELL LINES:

SUB CULTURING PROCEDURE FOR CELL LINES Check the cells for microbial contamination Add 1-2 ml trypsin/EDTA Incubate at 37C or at Rt for 5mins to cell detach Check the cells detachment using micoscope Cells are resuspended and glycerol stock is prepared for suspension lines

COMMON CONTAMINATIONS :

COMMON CONTAMINATIONS Fungal contamination Bacterial Contamination Yeast Contamination

MTT Assay:

MTT Assay Used to assess the viability and the proliferation of cells Colorimetric assay for measuring activity of enzymes that reduce MTT Determine cytotoxity of potential medicinal agents and toxic materials MTT- [3-(4, 5-Dimethylthiazol-2-yl)-2,5-Diphenyl tetrazolium bromide]

OBSERVATION:

OBSERVATION MCF-7 cell line MCF-7(Adherent)

RESULTS:

RESULTS The percentage of inhibition is measured by % cell survival: 100 – {(At-Ab) / (Ac-Ab)} × 100 Where: At = Absorbance of test Ab = Absorbance of blank Ac = Absorbance of control % cell inhibition: 100 - % cell survival

TABLE:

TABLE Activity of Euphorbia hirta on Hep-2 & MCF-7 % Inhibition Log Conc(mg) HEP-2 MC7-7 1.7 9 17 2 12 19 2.3 23 54 2.5 20 51

GRAPH:

GRAPH

CONCLUSION:

CONCLUSION Investigation of crude botanical extracts has revealed potent biological activities of variable level. Anti cancer potential of crude extracts described another screening strategy to select active extracts. Methanolic leaf extract of Euphorbia hirta has presented antitumor activities Present study revealed Euphorbia hirta for exhibiting anticancer properties on throat and breast cancer cell lines. Euphorbia hirta crude extract can further be fractionated to identify and isolate active components, which can provide important anticancer drugs in future.

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