In Vitro Apoptotic Activity of Endophytic Fungal Lectin Isolated from Endophyte, Aspergillus flavus of Viscum album on Human Breast Adenocarcinoma Cell Line (MCF-7Sadananda TS1, Govindappa M1*, Ramachandra YL2, Chandrappa CP1 and Umashankar T1
- *Corresponding Author:
- Govindappa M
Endophytic Natural Products Laboratory
Department of Biotechnology
Shridevi Institute of Engineering and Technology
Sira Road, Tumkur-572 106, Karnataka, India
Fax: 0816-2211752, 2212628
E-mail: [email protected]
Received Date: December 28, 2015; Accepted Date: January 11, 2016; Published Date: January 14, 2016
Citation: Sadananda TS, Govindappa M, Ramachandra YL, Chandrappa CP, Umashankar T (2016) In Vitro Apoptotic Activity of Endophytic Fungal Lectin Isolated from Endophyte, Aspergillus flavus of Viscum album on Human Breast Adenocarcinoma Cell Line (MCF-7). Metabolomics 6:162. doi: 10.4172/2153-0769.1000162
Copyright: © 2016 Sadananda TS, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Lectin was isolated and purified from the endophytic fungi Aspergillus flavus of Viscum album and it was a D-galactose and N-acetylgalactoseamine specific lectin capable of agglutinating A+ve blood group tested in hemeagglutination method. The lectin was confirmed a glycoprotein PAS staining on NATIVE-PAGE and it is having single band of 64 kDa protein (in SDS-PAGE) similar to plant lectin. The peptide sequences were identified from purified lectin 64 kDa protein band is shown. NVRFDLSGATSSSYK similar to Cucurbia pepo protein peptide sequence and it is a ribosome inactivating protein. Apoptotic activity was tested on human breast adenocarcinoma cancer (MCF-7) cell line and it is a time and dose dependent manner. Dose dependent inhibition of cancer cells was observed with IC50 values of 0.02 μg/ ml from Aspergillus flavus. MCF-7 cell death elicited by the lectin extract was found to be apoptotic in nature based the indication of nucleus condensation, shrinkage of nucleus membrane; membrane bebbling and also DNA fragmentation are the hallmark of apoptosis was observed in MCF-7 cells after staining with Hoechst 33342. Caspase mediated apoptosis was observed in lectin treated MCF-7 cells by cleaving fluorogenic peptide substrates, Ac-DEVD-AMC. The caspase-7 was observed as an effector in cleaving other caspases, further inhibition of caspase with an inhibitor provide clear evidence of caspase-7 mediated apoptosis in MCF-7 treated cells.