INTRODUCTION:

Ayurveda, a traditional Indian medicinal practice using plant drugs, has been effective in using these natural drugs from very early times and in preventing or suppressing various tumors with different treatment lines¹. In India, people of different ethnic groups live in different cultures, religious rites, food habits and a rich knowledge of traditional medicine. Cancer is a group of diseases caused by a loss of control over the cell cycle. Cancer is related to abnormal uncontrolled growth of the cells. Cancer is caused by both external factors (tobacco, chemicals, toxins, and infectious organisms) and internal factors (mutations inherited from genes, hormones, immune conditions, and mutations)². It poses a significant burden on public health in both developed and developing countries³. Anticancer activity is the effect of naturally occurring, synthetic or biological and chemical agents to reverse, inhibit or avoid cancer⁴. Several synthetic agents are used to treat the disease but they have their toxicity and hence the work is under way to investigate chemotherapeutic agents extracted from the plant⁵. Chemoprevention is recognized as a significant approach to malignancy management and recent studies have concentrated on identifying appropriate chemopreventive⁶. Breast cancer is the leading cause of death from cancer worldwide and the most common among women. Worldwide, there is a significant variation in breast cancer survival rates, where 80% of low-income countries in high-income countries are estimated to be below 40% for 5 years⁷. Natural products, particularly dietary substances, have been played role in developing new chemopreventive⁸. A paradigm focused on ethnobotanical and ethnopharmacological data would be more economical and effective in identifying potential anticancer molecules in any cancer clinical trials than mass screening of plant species. Natural products were considered essential sources of potential chemotherapeutic agents, and many anticancer drugs derived from natural sources⁹.

MATERIALS AND METHODS:

Plant material:

Psidiumguajava leaves has been collected from Karad, Maharashtra, India. Department of Botony, Yashwantrao Chavan College of Science, Karad has identified the plant and authenticated it.

Preparation of Psidiumguajavaleaf powder:

Fresh guava leaves were collected and air dried for 10 days. The dried leaves were then crushed into a blender and churned to form a coarse powder. The powder was collected in an air-tight jar, and stored away from sunlight in a cool, dry place.

Preparation of plant extract:

Extraction of malvastrumcoromandelianum was done by microwave extraction method further filtered and excess solvent present was evaporated and dried extract were collected and subjected for activity studies.

Chemicals:

The chemicals used in the present study was phosphate Buffered Saline (PBS) and tryphan blue dye from, Research lab, Mumbai. All the chemicals used were of analytical grade.

Cell lines and culture conditions:

MCF-7 cell line obtained from National Centre for Cell Sciences (NCCS), Pune. The cells were cultured at 37˚C in a humidified atmosphere of 5% CO₂ incubator.

Anticancer activity:

Tryphan blue exclusion method:

Place 50µl of cell suspension in cryo-vial. Add equal parts of 0.4% tryphan blue dye to the cell suspension to obtain a 1 to 2 dilution (example: 50µl of cells to µl of tryphan blue) and mix by pipetting up and down¹⁰ Incubate the mixture for less than three minutes at room temperature. If cells are counted after approximately five minutes, viability will be inaccurate due to the cell¹¹. With the cover slip already in place, fill one side of a hemocytometer counter with the cell suspension by placing the tip of the pipette at the notch. Typically, each side will take 10 to 20µl. Place the hemocytometer on the stage of a motic microscope and focus onto the cells. Each side of the hemocytometer contains multiple squares. Count all cells in each squares in each corner of the hemocytometer¹². Each large square contains 16 small squares. In each large square count cells that are on the border lines on two sides only. Keep track of the number of the blue cells seperatly as well as part of the complete number of cells^13-14.

RESULTS AND DISCUSSION:

Anticancer activity of ethyl acetate extract was tested using trypan blue exclusion method. 5-fluorouracil used as standard anticancer drug. By performing anticancer activity it has been noted that percent viability of Psidiumguajavaby comparing with standard drug shows significant activity. The results obtained by performing anticancer activity are listed in Table 1.

Table 1: Anticancer activity of Psidiumguajavaleaf extract against MCF-7 cell line:

Sr.No	Test subs	% viability
Sr.No	Test subs	10 mcg	50 mcg	100 mcg
1	Methanolic extract	83.82	81.33	77.03
2	Ethyl acetate extract	79.75	80.56	75.28
3	Ethanolic extract	86.18	82.48	80.92
4	Standard(5-FU)	87.52	84.12	81.24

CONCLUSION:

The aim of this study was to evaluate the anticancer activity of various leaf extracts of Psidiumguajava. I have tried to explore the in-vitro anticancer activity by Tryphan blue exclusion assay. The ethanol and methanol leaf extracts of Psidiumguajavahas shown potent anticancer activity MCF-7 cancer cell lines. However, the mechanism of the anticancer activity has not yet been fully elucidated and further research is needed to explore the molecular mechanism of this herbal plant.

ACKNOWLEDGEMENT:

I express my sincere thanks to Vice-principal Prof. Dr. S. K. Mohite for providing me all necessary facilities and valuable guidance extended to me.

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Received on 02.06.2020 Modified on 06.07.2020

Res. J. Pharma. Dosage Forms and Tech.2020; 12(4):298-300.

DOI: 10.5958/0975-4377.2020.00049.X