Botany - Research Publications

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    DNA barcoding and phylogeny based comparative evaluation of anti-cancer properties of Caulerpa (J V Lamouroux) spp. from Indian coasts
    (Central University of Punjab, 2019) Mehra, Richa; Bast, Felix and Singh, Sandeep
    A total of 15 Caulerpa samples were collected from Indian coasts and identified based on morphological and molecular data inferred from ITS, 18S, tufA and rbcL. Seven different species viz. C. scalpelliformis, C. racemosa, C. sertularioides, C. verticillata, C. taxifolia, and C. corynephora; and their geographical isolates were identified. Barcode data for these species was generated using aforementioned molecular markers and used for phylogenetic assessment. Phylogenetic trees using Bayesian inference (BI) and Maximum Likelihood (ML) function were generated for each molecular marker. tufA was found to be most suitable marker for the genus Caulerpa, resolving the species into 17 different lineages, with 15 corresponding to already known sections and 2 new lineages. Besides, a database named DbIndAlgae of Indian algae was generated and all the morphological as well as molecular data generated in this study is uploaded on the database. In addition, the phycochemical analysis revealed the presence of alkaloids, terpenoids, steroids, tannins, saponins, flavonoids, and phenols in different Caulerpa species. The selective cytotoxicity of methanolic extracts of Caulerpa (CMEs) was evaluated on MDA-MB-231, T47-D and H1299 cells, and the results revealed significant cytotoxicity of all species. C. racemosa KNY-254 and C. taxifolia TEN-158 were found to be most potent on MDA-MB-231 cells with IC50 value of 0.226 ± 0.004 and 0.246 ± 0.009 µg/µL. The mitochondrial membrane perturbation was revealed by JC-1 and apoptotic cell death was confirmed by Annexin V/FITC staining. CMEs also induced ROS in MDA-MB-231 cells as depicted by DHE, and increased activity of SOD, decreased activity of gluthatione reductase. The CMEs also exhibit anti-invasion activity and inhibited up to 71% migration across the artificially scratched wound in MDA-MB-231, w.r.t. untreated control cells. Moreover, chemical probing of C. racemosa KNY-254 by LC-MS analysis revealed six previously reported and six unreported molecules. The molecular docking analysis revealed weak to moderate interactions with all of the protein targets viz. Bcl2, AMPK, mTOR, BID, PERK, IGF-1R, PI3K, PTP1B and Akt2, known to play important role in cancer cell signaling. Additionally, a moderately positive correlation amongst the phylogeny and anti-cancer activity was observed suggesting that phylogeny might provide cues for anti-cancer activity, subject to further validations.
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    DNA barcode-based identification and comparative anti-cancer effects of different species of brown seaweed Sargassum C. Agardh of Indian coasts
    (Central University of Punjab, 2018) Bhushan, Satej; Bast, Felix & Singh, Sandeep
    Sargassum C. Agardh is a ubiquitous, multicellular brown seaweed that represents the most species-rich genus of the brown algal order Fucales, with more than 500 species reported worldwide. The present study aimed to identify different Sargassum isolates from India by DNA barcoding of mitochondrial (cox3), chloroplast (rbcL), and nuclear (18S) regions and further phylogenetic analyses. Total of 17 geographical isolates were collected across Indian coasts. Phylogeny reconstruction using Bayesian Inference was done which suggested congruency with known taxonomic hierarchy of Sargassum. Total of five different species were identified (S. portierianum, S. cymosum, S.aquifolium, S. ilicifolium, S. polycystum). In addition, comparative evaluation of anti-cancer potential of all the isolates was carried out and putative relationship between phylogeny and anticancer potential was established. MTT assay with 3 different cell lines showed cytotoxicity with IC50 as low as 0.167 ± 0.01, 0.243 ± 0.007, 0.25 ± 0.03 µg/µL in MDA-MB-231 (Breast Cancer), T-47D (Breast Cancer), H1299 (Lung Cancer) cells respectively, while no toxicity was observed with human peripheral blood mononuclear cells (hPBMCs). I was also able to isolate one lead aliphatic compound (SA1) whch was identified to be a polysaccharide using NMR spectroscopy. Similar to the extract, purified compound SA1 also showed anticancer activity. Further evaluations revealed that SA1 as well as the extracts interfere with the antioxidant defence components of cancer cells (SOD, Catalase, and GR) which results in the induction of mitochondrial death pathway at G1 phase (for extracts) as well as at G2M phase (for SA1). Extracts as well as SA1 were also able to inhibit cancer cell migration at sub IC50 doses. In addition, sub IC50 treatments lead to decreased colony formation compared to the control. Overall, our results show that these extracts as well as SA1 are able to target multiple properties of cancer