In-vitro Anti-cancer Activity of Endophytic Fungi Isolated from Ziziphus mauritiana in Cervical Cancer Cell Line

Abstract

Aim: The fungal crude extract of Trichoderma viride isolated from the medicinal plant of Ziziphus mauritiana was investigated for anti-cancer activity against HeLa cell line. Place and Duration of Study: The entire research work was done in the Department of Biotechnology, Mohammed Sathak College of Arts and Science, Chennai, India and Stellixir Biotech Pvt Ltd, Bangalore, Karnataka, between April 2019 to June 2019. Methodology: In MTT assay, cells were seeded in 96 well plates with concentrations of 2.5, 25, 50, 100, 200 μl/ml of fungal bioactive compound and the absorbance was measured at 570 nm. Apoptosis was carried out via flow cytometry using control Camptothecin and fungal extract concentration. Subsequently, cell cycle arrest was determined by 400 μl PI/RNAse staining buffer and further analyzed using PI solutions by flow cytometry. Finally, the damage of the nucleus observed under a Fluorescent microscope using DAPI staining. Results: MTT assay revealed an obtained fungal compound showed IC50 concentration against the HeLa cells at 23.57 μg/ml. Apoptosis analysis suggested under the M1 population the viable cells are low in mean Annexin V-FITC while M2 is the apoptotic cells have taken up the Annexin V-FITC and high in mean Annexin V-FITC fluorescence intensity. The cell cycle arrest has been noted by decreasing the percentage of arrest in the G0/G1 phase and increased in S and G2/M phase compared to controls. In the course of DAPI staining, it has been understood that IC50 23.57 μg/ml has an efficient nuclear dent after the treatment. Conclusion: In-vitro studies revealed that Trichoderma viride has promising cytotoxicity effect against the HeLa cell line. Hence the bioactive compounds of 3-beta-hydroxy urs-12-en-28-oic acid which is responsible for the treatment of cervical cancer. However in-vivo experiments need to be conceded out for further studies.