Exopolysaccharide from the mice ovarian bacterium Bacillus velezensis OM03 triggers caspase-3-dependent apoptosis in ovarian cancer cells

Sreejesh Pilakkavil Chirakkara Asha Abraham   

Open Access   

Published:  Feb 08, 2023

DOI: 10.7324/JAPS.2023.110355
Abstract

Exopolysaccharides (EPS) were isolated from mice ovarian microflora Bacillus velezensis OM03 to investigate their chemical properties and cytotoxic potential against human ovarian carcinoma cell lines PA-1 and SKOV-3. The structural analysis of EPS from OM03 using Fourier Transform Infra-Red spectroscopy, X-ray diffraction, Scanning Electron Microscopy, and Gas chromatography-Mass revealed that it is a novel heteropolysaccharide made of glucose and mannose units connected with α-1,4 and β-1,4 bonds. Attractively, the EPS inhibited the proliferation of SKOV-3 and PA-1 cancer cells in a concentration-dependent manner, with IC50 values of 620 and 238 μg/ml, respectively, according to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tests. Moreover, EPS-OM03 caused DNA fragmentation in PA-1 cell lines and boosted the expression of active caspase-3, both of which triggered apoptosis. Furthermore, the experiment with the chick embryo chorioallantoic membrane demonstrated that treatment with EPS-OM03 exhibits an in vivo antiangiogenic effect with an IC50 of 146 μg/ml. In conclusion, our work offers scientific support for the development and use of pharmaceuticals derived from the microbiome, and it suggests that the EPS from B. velezensis OM03 may be a lead molecule in the treatment of ovarian cancer.


Keyword:     Exopolysaccharides FTIR XRD GC-MS ovarian microbiome anti-ovarian cancer


Citation:

Sreejesh PC, Abraham A. Exopolysaccharide from the mice ovarian bacterium Bacillus velezensis OM03 triggers caspase- 3-dependent apoptosis in ovarian cancer cells. J Appl Pharm Sci, 2023. https://doi.org/10.7324/JAPS.2023.110355

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

HTML Full Text

Reference

Abdelhamid SA, Mohamed SS, Selim MS. Medical application of exopolymers produced by marine bacteria. Bull Natl Res Cent, 2020; 44:1-14. https://doi.org/10.1186/s42269-020-00323-x

Angelin J, Kavitha M. Exopolysaccharides from probiotic bacteria and their health potential. Int J Biol Macromol, 2020; 162:853-65. https://doi.org/10.1016/j.ijbiomac.2020.06.190

Ayyal NM, Abbas ZMA, Karim AJ, Abbas ZMA, Al-Salihi KA, Khalaf JM, Mahmood DD, Mohammed EA, Jumaa RS, Abdul-Majeed DI. Bacterial isolation from internal organs of rats (Rattus rattus) captured in Baghdad city of Iraq. Vet World, 2019; 12:119-25. https://doi.org/10.14202/vetworld.2019.119-125

Brewster WR, Burkett WC, Ko EM, Bae-Jump V, Nicole McCoy A, Keku TO. An evaluation of the microbiota of the upper reproductive tract of women with and without epithelial ovarian cancer. Gynecol Oncol Reports, 2022; 42:101017. https://doi.org/10.1016/j.gore.2022.101017

Brix N, Samaga D, Hennel R, Gehr K, Zitzelsberger H, Lauber K. The clonogenic assay: robustness of plating efficiency-based analysis is strongly compromised by cellular cooperation. Radiat Oncol, 2020; 15:1-12. https://doi.org/10.1186/s13014-020-01697-y

Cai G, Liu Y, Li X, Lu J. New levan-type exopolysaccharide from Bacillus amyloliquefaciens as an antiadhesive agent against enterotoxigenic Escherichia coli. J Agric Food Chem, 2019; 67:8029-34. https://doi.org/10.1021/acs.jafc.9b03234

Cao C, Liu Y, Li Y, Zhang Y, Zhao Y, Wu R, Wu HJ. Structural characterization and antioxidant potential of a novel exopolysaccharide produced by Bacillus velezensis SN-1 from spontaneously fermented Da- Jiang. Glycoconj J, 2020; 37:307-17. https://doi.org/10.1007/s10719-020-09923-1

Castellane TCL, Otoboni AMMB, de MacedoLemos EG. Characterization of exopolysaccharides produced by Rhizobia species. Rev Bras Ciênc Solo, 2015; 39:1566-75. https://doi.org/10.1590/01000683rbcs20150084

Chen L, Zeng Y, Zhou SF, Chen L, Zeng Y, Zhou SF. Role of apoptosis in cancer resistance to chemotherapy. Current understanding of apoptosis-Program Cell Death, IntechOpen, Paris, France, 2018; 2018:126-36. https://doi.org/10.5772/intechopen.80056

Cheng H, Wang Z, Cui L, Wen Y, Chen X, Gong F, Yi H. Opportunities and challenges of the human microbiome in ovarian cancer. Front Oncol, 2020; 10:163. https://doi.org/10.3389/fonc.2020.00163

Chirakkara SP, Abraham A. Bacillus strains from the ovaries of Swiss albino mice (Mus musculus): deciphering of probiotic potential through an in vitro approach. Biomedicine, 2022; 42:1200-8. https://doi.org/10.51248/.v42i6.2302

Gangalla R, Gattu S, Palaniappan S, Ahamed M, Macha B, Thampu RK, Fais A, Cincotti A, Gatto G, Dama M, Kumar A. Structural characterisation and assessment of the novel Bacillus amyloliquefaciens RK3 exopolysaccharide on the improvement of cognitive function in Alzheimer's disease mice. Polymers (Basel), 2021; 13(17):2842. https://doi.org/10.3390/polym13172842

Guo R, Chen M, Ding Y, Yang P, Wang M, Zhang H, He Y, Ma H. Polysaccharides as potential anti-tumor biomacromolecules -a review. Front Nutr, 2022; 9:838179. https://doi.org/10.3389/fnut.2022.838179

Hong T, Yin JY, Nie SP, Xie MY. Applications of infrared spectroscopy in polysaccharide structural analysis: progress, challenge and perspective. Food Chem X, 2021; 12:100168. https://doi.org/10.1016/j.fochx.2021.100168

Hu M, Cui N, Bo Z, Xiang F. Structural determinant and its underlying molecular mechanism of STPC2 related to anti-angiogenic activity. Mar Drugs, 2017; 15(2):458. https://doi.org/10.3390/md15020048

Hu XM, Li ZX, Lin RH, Shan JQ, Yu QW, Wang RX, Liao LS, Yan WT, Wang Z, Shang L, Huang Y, Zhang Q, Xiong K. Guidelines for regulated cell death assays: a systematic summary, a categorical comparison, a prospective. Front Cell Dev Biol, 2021; 9:368. https://doi.org/10.3389/fcell.2021.634690

Jung MY, Kang HJ, Moon A. Capsaicin-induced apoptosis in SK-HEP-1 hepatocarcinoma cells involves bcl-2 downregulation and caspase-3 activation. Cancer Lett, 2001; 165:139-45. https://doi.org/10.1016/S0304-3835(01)00426-8

Kavitha M, Raja M, Perumal P. Evaluation of probiotic potential of Bacillus spp. isolated from the digestive tract of freshwater fish Labeo calbasu (Hamilton, 1822). Aquac Rep, 2018; 11:59-69. https://doi.org/10.1016/j.aqrep.2018.07.001

Koroth J, Nirgude S, Tiwari S, Gopalakrishnan V, Mahadeva R, Kumar S, Karki SS, Choudhary B. Investigation of anti-cancer and migrastatic properties of novel curcumin derivatives on breast and ovarian cancer cell lines. BMC Complement Altern Med, 2019; 19:1-16. https://doi.org/10.1186/s12906-019-2685-3

Krishnamurthy M, Jayaraman Uthaya C, Thangavel M, Annadurai V, Rajendran R, Gurusamy A. Optimization, compositional analysis, and characterization of exopolysaccharides produced by multi-metal resistant Bacillus cereus KMS3-1. Carbohydr Polym, 2020; 227:115369. https://doi.org/10.1016/j.carbpol.2019.115369

Li M, Liu Y, Zhang H, Liu Y, Wang W, You S, Hu X, Song M, Wu R, Wu J. Anti-cancer potential of polysaccharide extracted from Polygonatum sibiricum on HepG2 cells via cell cycle arrest and apoptosis. Front Nutr, 2022; 9:1-12. https://doi.org/10.3389/fnut.2022.938290

Li N, Wang C, Georgiev MI, Bajpai VK, Tundis R, Simal-Gandara J, Lu X, Xiao J, Tang X, Qiao X. Advances in dietary polysaccharides as anticancer agents: structure-activity relationship. Trends Food Sci Technol, 2021; 111:360-77. https://doi.org/10.1016/j.tifs.2021.03.008

Mahgoub AM, Mahmoud MG, Selim MS, El Awady ME. Exopolysaccharide from marine Bacillus velezensis MHM3 induces apoptosis of human breast cancer MCF-7 cells through a mitochondrial pathway. Asian Pac J Cancer Prev, 2018; 19:1957-63.

Mathivanan K, Chandirika JU, Mathimani T, Rajaram R, Annadurai G, Yin H. Production and functionality of exopolysaccharides in bacteria exposed to a toxic metal environment. Ecotoxicol Environ Saf, 2021; 208:111567. https://doi.org/10.1016/j.ecoenv.2020.111567

Moghannem SAM, Farag MMS, Shehab AM, Azab MS. Exopolysaccharide production from Bacillus velezensis KY471306 using statistical experimental design. Braz J Microbiol, 2018; 49:452-62. https://doi.org/10.1016/j.bjm.2017.05.012

Mohanta M, Khanam S, Islam MS, Mohanta MK. Isolation, characterization and identification of bacterial isolates from the poultry environment at Rajshahi Metropolis, Bangladesh. Artic J Entomol Zool Stud, 2017; 5:918-26.

Mohite BV, Koli SH, Rajput JD, Patil VS, Agarwal T, Patil SV. Production and characterization of multifacet exopolysaccharide from an agricultural isolate, Bacillus subtilis. Biotechnol Appl Biochem, 2019; 66:1010-23. https://doi.org/10.1002/bab.1824

Moradali MF, Rehm BHA. Bacterial biopolymers: from pathogenesis to advanced materials. Nat Rev Microbiol, 2020; 18(4):195-210. https://doi.org/10.1038/s41579-019-0313-3

Naik M, Brahma P, Dixit M. A cost-effective and efficient chick Ex-Ovo CAM assay protocol to assess angiogenesis. Methods Protoc, 2018; 1:1-9. https://doi.org/10.3390/mps1020019

Nasr SA, Saad AAEM. Evaluation of the cytotoxic anticancer effect of polysaccharide of Nepeta septemcrenata. Beni-Suef Univ J Basic Appl Sci,2021; 10:1-11. https://doi.org/10.1186/s43088-021-00135-6

Nguyen PT, Nguyen TT, Vo TNT, Nguyen TTX, Hoang QK, Nguyen HT. Response of Lactobacillus plantarum VAL6 to challenges of pH and sodium chloride stresses. Sci Rep, 2021; 11:1-9. https://doi.org/10.1038/s41598-020-80634-1

Obeng E. Apoptosis (Programmed cell death) and its signals-a review. Braz J Biol, 2021; 81:1133-43. https://doi.org/10.1590/1519-6984.228437

Oerlemans MMP, Akkerman R, Ferrari M, Walvoort MTC, de Vos P. Benefits of bacteria-derived exopolysaccharides on gastrointestinal microbiota, immunity and health. J Funct Foods, 2021; 76:104289. https://doi.org/10.1016/j.jff.2020.104289

Oleksy-Sobczak M, Klewicka E. Optimization of media composition to maximize the yield of exopolysaccharides production by Lactobacillus rhamnosus strains. Probiotics Antimicrob Proteins, 2020; 12:774-83. https://doi.org/10.1007/s12602-019-09581-2

Paudel KR, Dharwal V, Patel VK, Galvao I, Wadhwa R, Malyla V, Shen SS, Budden KF, Hansbro NG, Vaughan A, Yang IA, Kohonen- Corish MRJ, Bebawy M, Dua K, Hansbro PM. Role of lung microbiome in innate immune response associated with chronic lung diseases. Front Med, 2020; 7:554. https://doi.org/10.3389/fmed.2020.00554

Pu X, Storr SJ, Zhang Y, Rakha EA, Green AR, Ellis IO, Martin SG. Caspase-3 and caspase-8 expression in breast cancer: caspase-3 is associated with survival. Apoptosis, 2017; 22:357-68. https://doi.org/10.1007/s10495-016-1323-5

Ravi G, Sampath G, Srinivas B, Sarika K, Govindarajan RK, Ameen F, Alwakeel S, Thampu RK. Erratum: optimization and characterization of exopolysaccharide produced by Bacillus aerophilus RK1 and its in vitro antioxidant activities. J King Saud Univ Sci, 2021; 33(5):101571. https://doi.org/10.1016/j.jksus.2021.101571

Saravanan C, Shetty PKH. Isolation and characterization of exopolysaccharide from Leuconostoc lactis KC117496 isolated from idli batter. Int J Biol Macromol, 2016; 90:100-6. https://doi.org/10.1016/j.ijbiomac.2015.02.007

Schmid J, Sieber V, Rehm B. Bacterial exopolysaccharides: biosynthesis pathways and engineering strategies. Front Microbiol, 2015; 6:496. https://doi.org/10.3389/fmicb.2015.00496

Smith BC. The C=O bond, part III: carboxylic acids. Spectroscopy, 2018; 33:14-20-14-20.

Sun B, Straubinger RM, Lovell JF. Current taxane formulations and emerging cabazitaxel delivery systems. Nano Res, 2018; 11:5193-218. https://doi.org/10.1007/s12274-018-2171-0

Trivedi S, Patel K, Belgamwar V, Wadher K. Functional polysaccharide lentinan: role in anti-cancer therapies and management of carcinomas. Pharmacol Res Mod Chinese Med, 2022; 2:100045. https://doi.org/10.1016/j.prmcm.2022.100045

Vidhyalakshmi R, Valli Nachiyar C, Narendra Kumar G, Sunkar S. Bacillus circulans exopolysaccharide: production, characterization and bioactivities. Int J Biol Macromol, 2016; 87:405-14. https://doi.org/10.1016/j.ijbiomac.2016.02.001

Vigneswara V, Ahmed Z. The role of caspase-2 in regulating cell fate. Cells, 2020; 9(5):1259. https://doi.org/10.3390/cells9051259

Vinothkanna A, Sathiyanarayanan G, Rai AK, Mathivanan K, Saravanan K, Sudharsan K, Kalimuthu P, Ma Y, Sekar S. Exopolysaccharide produced by probiotic Bacillus albus DM-15 isolated from ayurvedic fermented dasamoolarishta: characterization, antioxidant, and anticancer activities. Front Microbiol, 2022; 13:213. https://doi.org/10.3389/fmicb.2022.832109

Wang J, Salem DR, Sani RK. Two new exopolysaccharides from a thermophilic bacterium Geobacillus sp. WSUCF1: characterization and bioactivities. N Biotechnol, 2021; 61:29-39. https://doi.org/10.1016/j.nbt.2020.11.004

Wang L, Wang Y, Li Q, Tian K, Xu L, Liu G, Guo C. Exopolysaccharide, isolated from a novel strain Bifidobacterium breve lw01 possess an anticancer effect on head and neck cancer - genetic and biochemical evidences. Front Microbiol, 2019; 10:1-10. https://doi.org/10.3389/fmicb.2019.01044

Wei L, Ma F, Du C. Application of FTIR-PAS in rapid assessment of rice quality under climate change conditions. Foods, 2021; 10:159. https://doi.org/10.3390/foods10010159

Wiercigroch E, Szafraniec E, Czamara K, Pacia MZ, Majzner K, Kochan K, Kaczor A, Baranska M, Malek K. Raman and infrared spectroscopy of carbohydrates: a review. Spectrochim Acta Part A Mol Biomol Spectrosc, 2017; 185:317-35. https://doi.org/10.1016/j.saa.2017.05.045

Wu J, Lin C, Chen X, Pan N, Liu Z. Polysaccharides isolated from Bangia fuscopurpurea induce apoptosis and autophagy in human ovarian cancer A2780 cells. Food Sci Nutr, 2021; 9:6707-19. https://doi.org/10.1002/fsn3.2621

Zhang L, Yi H. Potential antitumor and anti-inflammatory activities of an extracellular polymeric substance (EPS) from Bacillus subtilis isolated from a housefly. Sci Rep, 2022; 12:1-10. https://doi.org/10.1038/s41598-022-05143-9

Zhang Y, Dai X, Jin H, Man C, Jiang Y. The effect of optimized carbon source on the synthesis and composition of exopolysaccharides produced by Lactobacillus paracasei. J Dairy Sci, 2021; 104:4023-32. https://doi.org/10.3168/jds.2020-19448

Zhou X, Kandalai S, Hossain F, Zheng Q. Tumor microbiome metabolism: a game changer in cancer development and therapy. Front Oncol, 2022;12:3680. https://doi.org/10.3389/fonc.2022.933407

Article Metrics

0 Absract views 3 PDF Downloads 3 Total views

   Abstract      Pdf Download

Related Search

By author names

Citiaion Alert By Google Scholar

Name Required
Email Required Invalid Email Address

Comment required