Ramya, N (2014) Evaluation of Anticancer Activity of Various Extract of Seeds of Trachyspermum Ammi (L.) Sprangue Using Breast Cancer Celll Lines by Invitro Methods. Masters thesis, Institute of Pharmacology, Madras Medical College, Chennai.
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Abstract
AIM AND OBJECTIVES: The present study to evaluate Phyto chemical properties of the various extracts of seeds of Trachyspermum ammi. To evaluate the IC50 concentration of ethanolic extracts of Trachyspermum ammi. To determine the apoptotic activity of ethanolic extracts of Trachyspermum ammi. To determine the gene expression of cancer DNA markers in MCF-7 Cell line treated with ethanolic extracts of Trachyspermum ammi. Breast cancer is malignant neoplasm occurring in females. It accounts for 23% of newly occurring cancer worldwide and represents 13.7% of all cancer deaths. In both developed and developing countries, breast cancer mortality and incidence rates are projected to rising continuously. In spite of advancing different types of treatments, the rate of breast cancer has not decreased yet. The report, by NCEPOD has shown that more than 1 in 4 patients died from the side effect rather than from cancer and patients are suffering from treatment related toxicity. Because of dangerous side effects and high cost of chemotherapy, usage of herbal medicine is on increased in large number of population. In search of new compound to treat breast cancer with fewer side effects, researchers have been evaluating number of medicinal plants. Since the medicinal plants have phyto constituents, which protects the plant from oxidative damages and the same phytoconstituents which when used in humans may have the same role. They have wide range of action such as antitumor, antiviral, antibacterial, antimutagenic etc and they may act in different stage of malignant tumor by protecting the DNA from oxidative damages. They inactivate carcinogen by inhibiting the expression of mutagenic genes; they also inactivate the enzymes with activate the systems responsible for the detoxification of xenobiotics (Bravo, 1998) Trachyspermum ammi has been reported to show a number of biological activities in traditional medicine and it has been used as digestive, carminative, laxative, diuretic, stimulant and antispasmodics. Several investigation in the seeds of Trachyspermum ammi have been indicated to posses many activities like potent antibacterial, antimicrobial, anthelmintics, hypolipidemic, etc. Phyto Chemical Analysis The phyto chemical analysis of ethanolic extract of seeds of T. ammi revealed the presence of flavanoids, terepenoids, tannis, alkaloids, steroids, glycosides, carbhohydrates. The acetone and aqueous extracts didn’t show the presence of flavanoid and terepenoids. The ethanolic extract revealed the presence of flavanoids and terepenoids which shows that it may posses anticancer activity and destroy cancer cells. MTT Assay: The MTT assay was carried out to determine the IC50 concentration of ethanolic extract of Trachyspermum ammi.The effective IC50 of ethanolic extract was found to be 125 μg/ml. The effective IC50 concentration of ethanolic extract of Trachyspermum ammi and standard 5-fluorouracil was chosen for further studies. Light Microscopy: Control did not show apoptosis, but cells treated with of ethanolic extract of Trachyspermum ammi showed typical morphological features of apoptosis like destruction of cells, reduction of MCF-7 cell population, reduction of cell volume, loss of integrity of membrane which resulted in crooked and vesicle shape of the membrane and chromatin condensation. Fluorescence Microscopy: Apoptotic effects of IC50 concentration of ethanolic extract of Trachyspermum ammi treated MCF-7 cells were further confirmed with the help of fluorescence microscopy. Control did not reveal any apoptotic activity. Cells treated with of ethanolic extract of Trachyspermum ammi showed apoptotic activity. The presence of apoptotic activity shows ethanolic extract of Trachyspermum ammi may have anticancer activity. On comparing with control group, both 5 Fluorouracil and ethanolic extract of Trachyspermum ammi has produced DNA fragmentation in MCF-7 cells. The fragmentation of DNA indicates that apoptotis has occured. This shows ethanolic extract of Trachyspermum ammi posses apoptotic activity. Gene Expression Cells treated with 5 Fluorouracil, expression levels of p53 was found to be increased and similar increase was observed in cells treated with ethanolic extract of Trachyspermum ammi. Expression levels of Bcl-2 were decreased in cells treated with 5 Fluorouracil. Cells treated with ethanolic extract of Trachyspermum ammi shows a decrease in expression levels of Bcl-2 slightly lower than 5 Fluorouracil. Expression levels of p53 and Bcl-2 indicated that ethanolic extract of Trachyspermum ammi has anticancer activity. CONCLUSION: From the present study it may be concluded that, The phyto chemical investigation revealed the presence of flavanoids, tannins, trepenoids, steroids, glycosides, alkaloids. By MTT assay, The IC50 value of ethanolic extract of Trachyspermum ammi was found to be 125μg/ml. The ethanolic extract of Trachyspermum ammi posses apoptotic activity. The gene expression of cancer DNA markers in MCF-7 Cell line treated with ethanolic extracts of Trachyspermum ammi showed increase in p53 expression and decrease in Bcl2 expression.
| Item Type: | Thesis (Masters) |
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| Uncontrolled Keywords: | Seeds; Trachyspermum Ammi (L.); Sprangue; Breast Cancer Celll Lines; Invitro Methods |
| Subjects: | PHARMACY > Pharmacology |
| Depositing User: | Ravindran C |
| Date Deposited: | 23 Oct 2017 05:24 |
| Last Modified: | 23 Oct 2017 05:24 |
| URI: | http://repository-tnmgrmu.ac.in/id/eprint/3766 |
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