Dr.Swetha Madamala

MVJ College of Engineering (Karnataka), India

Title: INVITRO CYTOTOXICITY OF URSOLIC ACID, 6 METHYL CHROMONE HYDRATE AND GYMNEMIC ACID ON MCF7 ANDA549 CELL LINES

---

Biography

Dr Swetha Madamala recently did her Doctoral studies at JNTUA. She did her doctoral studies on “Phytochemical Investigation, Method Development and Validation of Anti-Cancer Herbal Plants Gymnema Slyvestrae, Morinda Citrifolia and Aegle Marmeleous Using RP HPLC Techniques.  Her area of research interest in HPLC techniques, Herbal Chemistry, Nanotechnology. she is having 10 years’ experience in teaching graduate students. She attended various conferences on national and international and published a few papers on reputed journals, she is an active member in various professional bodies. Currently working as Assistant  Professor in MVJ College of Engineering.

Abstract

In modern medicine, chemotherapy, radiotherapy, and surgery are the major existing modes of

treatments. The toxicity and severe adverse effects associated with cancer chemotherapy and radio therapy create new avenues for discovering and developing nontoxic agents for prophylaxis, mitigation, and treatment of cancer. One of the best approaches in searching for novel anticancer agents from plant resources is selection of plants based on ethnomedical practices and testing their efficacy and safety considering modern science. In the past two decades, systemic ethno botanical documentation has been prioritized in India and recent studies indicate that plants used by herbal healers have been scientifically shown to possess chemotherapeutic value. This added to deep belief that these treatments are safe because they are” natural” and fit into the image of a gentle and therefore, harmless alternative to conventional medicine and hence are staging a comeback and herbal renaissance in treating cancer is happening all over the world. as most of the blockbuster agents are from botanicals.

Ursolic Acid, 6Methyl chromone hydrate, Gynmemic Acid were tested for invitro cytotoxicity, using A549 cell Lines and MCF7 cell Lines by using MTT assay.

The monolayer cell culture of the employed cell lines was trypsinized, cell count was adjusted to100,000 cells/ml with MEM containing 10% FBS. To each well of the 96 well microtiter plate, 100 µL of the diluted cell suspension was added. After 24 h, when a partial monolayer was formed, the supernatant was flicked off, monolayer washed once with medium and 100 µLof different test concentrations of test substances were added on to the partial monolayer. 200 µL of cells (A549 &MCF7) without test substance treatment were taken as control. Each sample was replicated thrice, and cells were incubated at 37^o C for 72 h in a humidified 5% CO₂ incubator and microscopic examination was carried out and observations were noted every 24 h interval.

                 After 72 h incubation, the drug solutions in the wells were discarded and 50 L of MTT in PBS was added to each well. The plates were gently shaken and incubated for 3 h at 37^o C in 5% CO₂ atmosphere. The supernatant was removed and 100µL of propanol was added and the plates were gently shaken to solubilize the formed formazan. The absorbance was measured using a microplate reader at a wavelength of 540 nm. The percentage growth inhibition was calculated using the standard formula and concentration of test substances needed to inhibit cell growth by 50% (CTC₅₀) values was generated from the dose-response curves for each cell and the percentage viability (CV)was calculated manually using formula:

CV=   Average absorbance of treated drug wells- Absorbance of Blank     X                                 100%

                       Average absorbance of control drug wells- Absorbance of Blank      Blank cells contain media alone with no plating of cells and control group cells are nothing but untreated cells.

                                    % Cytotoxicity = 100- % of cell viability

    The mean of absorbance values that are lower than control group indicates reduction in cell viability. Conversely, a higher mean absorbance indicates increase in cell proliferation.

A dose response curve was plotted to enable the calculation of the concentrations that kill 50%

 of the A549 and MCF7 cells. (IC₅₀)