Evaluation of the anticarcinogenic potential of the endophyte, Streptomyces sp. LRE541 isolated from Lilium davidii var. unicolor (Hoog) Cotton

Microbial Cell Factories - 2021
Aiai Ma1, Kan Jiang2, Bin Chen2, Shasha Chen3, Xing-e Qi4, Huanjun Lu5, Junlin Liu5, Xuan Zhou1, Tieren Gao1, Jinhui Li4, Changming Zhao1
1Yuzhong Mountain Ecosystem Field Observation and Research Station, Lanzhou University, Lanzhou, China
2College of Agronomy, Gansu Agricultural University, Lanzhou, China
3Department of Animal and Biomedical Sciences, School of Life Sciences, Lanzhou University, Lanzhou, China
4State Key Laboratory of Grassland Agro-ecosystems, School of Life Sciences, Lanzhou University, Lanzhou, China
5Life Science and Engineering College of Northwest University for Nationalities, Lanzhou, China

Tóm tắt

Abstract Background

Endophytic actinomycetes, as emerging sources of bioactive metabolites, have been paid great attention over the years. Recent reports demonstrated that endophytic streptomycetes could yield compounds with potent anticancer properties that may be developed as chemotherapeutic drugs.

 Results

Here, a total of 15 actinomycete-like isolates were obtained from the root tissues of Lilium davidii var. unicolor (Hoog) Cotton based on their morphological appearance, mycelia coloration and diffusible pigments. The preliminary screening of antagonistic capabilities of the 15 isolates showed that isolate LRE541 displayed antimicrobial activities against all of the seven tested pathogenic microorganisms. Further in vitro cytotoxicity test of the LRE541 extract revealed that this isolate possesses potent anticancer activities with IC50 values of 0.021, 0.2904, 1.484, 4.861, 6.986, 8.106, 10.87, 12.98, and 16.94 μg/mL against cancer cell lines RKO, 7901, HepG2, CAL-27, MCF-7, K562, Hela, SW1990, and A549, respectively. LRE541 was characterized and identified as belonging to the genus Streptomyces based on the 16S rRNA gene sequence analysis. It produced extensively branched red substrate and vivid pink aerial hyphae that changed into amaranth, with elliptic spores sessile to the aerial mycelia. To further explore the mechanism underlying the decrease of cancer cell viability following the LRE541 extract treatment, cell apoptosis and cell cycle arrest assays were conducted in two cancer cell lines, RKO and 7901. The result demonstrated that LRE541 extract inhibited cell proliferation of RKO and 7901 by causing cell cycle arrest both at the S phase and inducing apoptosis in a dose-dependent manner. The chemical profile of LRE541 extract performed by the UHPLC-MS/MS analysis revealed the presence of thirty-nine antitumor compounds in the extract. Further chemical investigation of the LRE541 extract led to the discovery of one prenylated indole diketopiperazine (DKP) alkaloid, elucidated as neoechinulin A, a known antitumor agent firstly detected in Streptomyces; two anthraquinones 4-deoxy-ε-pyrromycinone (1) and epsilon-pyrromycinone (2) both displaying anticancer activities against RKO, SW1990, A549, and HepG2 with IC50 values of 14.96 ± 2.6 − 20.42 ± 4.24 μg/mL for (1); 12.9 ± 2.13, 19.3 ± 4.32, 16.8 ± 0.75, and 18.6 ± 3.03 μg/mL for (2), respectively.

 Conclusion

Our work evaluated the anticarcinogenic potential of the endophyte, Streptomyces sp. LRE541 and obtained one prenylated indole diketopiperazine alkaloid and two anthraquinones. Neoechinulin A, as a known antitumor agent, was identified for the first time in Streptomyces. Though previously found in Streptomyces, epsilon-pyrromycinone and 4-deoxy-ε-pyrromycinone were firstly shown to possess anticancer activities.

 Graphical Abstract

Từ khóa


Tài liệu tham khảo

Sara C, Maria P, Maeve K, Graham P, Denis C. Innovative technologies changing cancer treatment. Cancers. 2018;10:208.

Holohan C, Schaeybroeck SV, Longley DB, Johnston PG. Cancer drug resistance: an evolving paradigm. Nat Rev Cancer. 2013;13:714–26. https://doi.org/10.1038/nrc3599.

Hussain S, Singh A, Nazir SU, Tulsyan S, Khan A, Kumar R, Bashir N, Tanwar P, Mehrotra R. Cancer drug resistance: a fleet to conquer. J Cell Biochem. 2019;120:14213–25. https://doi.org/10.1002/jcb.28782.

Kerr JF, Winterford CM, Harmon BV. Apoptosis. Its significance in cancer and cancer therapy. Cancer. 1994;73:2013–26. https://doi.org/10.1002/1097-0142(19940415)73:8%3c2013::aid-cncr2820730802%3e3.0.co;2-j.

Cao K, Tait SW. Apoptosis and cancer: force awakens, phantom menace, or both? Int Rev Cell Mol Biol. 2018;337:135–52. https://doi.org/10.1016/bs.ircmb.2017.12.003.

Campbell KJ, Tait SW. Targeting BCL-2 regulated apoptosis in cancer. Open Biol. 2018;8:180002. https://doi.org/10.1098/rsob.180002.

Hayakawa Y, Kuriyama T, Ibusuki Y, Kimata S. Pyroxazone, a new neuroprotective compound from Streptomyces sp. RAN54. J Antibiot. 2018;71:962–4. https://doi.org/10.1038/s41429-018-0085-4.

Tan TH, Chan KG, Pusparajah P, Yin WF, Khan TM, Lee LH, Goh BH. Mangrove derived Streptomyces sp. MUM265 as a potential source of antioxidant and anticolon-cancer agents. BMC Microbiol. 2019;19:38–44. https://doi.org/10.1186/s12866-019-1409-7.

Bieber B, Nüske J, Ritzau M, Grafe U. Alnumycin a new naphthoquinone antibiotic produced by an endophytic Streptomyces sp. J Antibiot. 1998;51:381–2. https://doi.org/10.7164/antibiotics.51.381.

Ser HL, Palanisamy UD, Yin WF, Chan KG, Goh BH, Lee LH. Streptomyces malaysiense sp. nov.: a novel Malaysian mangrove soil actinobacterium with antioxidative activity and cytotoxic potential against human cancer cell lines. Sci Rep. 2016;6:24247–59. https://doi.org/10.1038/srep24247.

Remsing LL, González AM, Nur-e-Alam M, Fernández-Lozano MJ, Braña AF, Rix U, Oliveira MA, Méndez C, Salas JA, Rohr J. Mithramycin SK, a novel antitumor drug with improved therapeutic index, mithramycin SA, and demycarosyl-mithramycin SK: three new products generated in the mithramycin producer Streptomyces argillaceus through combinatorial biosynthesis. J Am Chem Soc. 2003;125:5745–53. https://doi.org/10.1021/ja034162h.

Taechowisan T, Lu C, Shen Y, Lumyong S. Antitumor activity of 4-arylcoumarins from endophytic Streptomyces aureofaciens CMUAc130. J Cancer Res Ther. 2007;3:86–91. https://doi.org/10.4103/0973-1482.34685.

Zhang R, Han X, Xia Z, Luo X, Wan C, Zhang L. Streptomyces luozhongensis sp. nov., a novel actinomycete with antifungal activity and antibacterial activity. Antonie Van Leeuwenhoek. 2017;110:195–203. https://doi.org/10.1007/s10482-016-0790-6.

Wang L, Qiu P, Long XF, Zhang S, Zeng ZG, Tian YQ. Comparative analysis of chemical constituents, antimicrobial and antioxidant activities of ethyl acetate extracts of Polygonum cuspidatum and its endophytic actinomycete, Streptomyces sp. A0916. Chin J Nat Med. 2016;14:117–23. https://doi.org/10.1016/s1875-5364(16)60004-3.

Perez Baz J, Canedo LM, Fernandez Puentes JL, Silva Elipe MV. Thiocoraline, a novel depsipeptide with antitumor activity produced by a marine Micromonospora. II. Physico-chemical properties and structure determination. J Antibiot. 1997;50:738–41. https://doi.org/10.7164/antibiotics.50.738.

Schneider K, Keller S, Wolter FE, Roglin L, Beil W, Seitz O, Nicholson G, Bruntner C, Riedlinger J, Fiedler HP, Sussmuth RD. Proximicins A, B, and C-antitumor furan analogues of netropsin from the marine actinomycete Verrucosispora induce upregulation of p53 and the cyclin kinase inhibitor p21. Angew Chem Int Ed Engl. 2008;47:3258–61. https://doi.org/10.1002/anie.200705295.

Golinska P, Wypij M, Agarkar G, Rathod D, Dahm H, Rai M. Endophytic actinobacteria of medicinal plants: diversity and bioactivity. Antonie Van Leeuwenhoek. 2015;108:267–89. https://doi.org/10.1007/s10482-015-0502-7.

Khieu T-N, Liu M-J, Nimaichand S, Quach N-T, Chu-Ky S, Phi Q-T, Vu T-T, Nguyen T-D, Xiong Z, Prabhu DM, Li W-J. Characterization and evaluation of antimicrobial and cytotoxic effects of Streptomyces sp. HUST012 isolated from medicinal plant Dracaena cochinchinensis Lour. Front Microbiol. 2015;6:574–574. https://doi.org/10.3389/fmicb.2015.00574.

Hui H, Jin H, Li X, Yang X, Cui H, Xin A, Zhao R, Qin B. Purification, characterization and antioxidant activities of a polysaccharide from the roots of Lilium davidii var. unicolor Cotton. Int J Biol Macromol. 2019;135:1208–16. https://doi.org/10.1016/j.ijbiomac.2019.06.030.

Barka EA, Vatsa P, Sanchez L, Gaveau-Vaillant N, Jacquard C, Meier-Kolthoff JP, Klenk HP, Clément C, Ouhdouch Y, van Wezel GP. Taxonomy, physiology, and natural products of actinobacteria. Microbiol Mol Biol Rev. 2016;80:1–43. https://doi.org/10.1128/mmbr.00019-15.

Gali-Muhtasib H, Hmadi R, Kareh M, Tohme R, Darwiche N. Cell death mechanisms of plant-derived anticancer drugs: beyond apoptosis. Apoptosis. 2015;20:1531–62. https://doi.org/10.1007/s10495-015-1169-2.

Kaufmann SH, Earnshaw WC. Induction of apoptosis by cancer chemotherapy. Exp Cell Res. 2000;256:42–9.

Pettit GR, Hogan F, Xu J-P, Tan R, Nogawa T, Cichacz Z, Pettit RK, Du J, Ye Q-H, Cragg GM, Herald CL, Hoard MS, Goswami A, Searcy J, Tackett L, Doubek DL, Williams L, Hooper JNA, Schmidt JM, Chapuis J-C, Tackett DN, Craciunescu F. Antineoplastic agents. 536. New sources of naturally occurring cancer cell growth inhibitors from marine organisms, terrestrial plants, and microorganisms. J Nat Prod. 2008;71:438–44. https://doi.org/10.1021/np700738k.

Wijesekara I, Li YX, Vo TS, Ta QV, Ngo DH, Kim SK. Induction of apoptosis in human cervical carcinoma HeLa cells by neoechinulin A from marine-derived fungus Microsporum sp. Process Biochem. 2013;48:68–72.

Deeble VJ, Lindley HK, Fazeli MR, Cove JH, Baumberg S. Depression of streptomycin production by Streptomyces griseus at elevated growth temperature: studies using gene fusions. Microbiology. 1995;141(Pt 10):2511–8. https://doi.org/10.1099/13500872-141-10-2511.

Srivibool R, Kurakami K, Sukchotiratana M, Tokuyama S. Coastal soil actinomycetes: thermotolerant strains producing N-acylamino acid racemase. ScienceAsia. 2004;30:123–6.

Basilio A, González I, Vicente MF, Gorrochategui J, Cabello A, González A, Genilloud O. Patterns of antimicrobial activities from soil actinomycetes isolated under different conditions of pH and salinity. J Appl Microbiol. 2003;95:814–23. https://doi.org/10.1046/j.1365-2672.2003.02049.x.

Zakalyukina YV, Zenova GM, Zvyagintsev DG. Peculiarities of growth and morphological differentiation of acidophilic and neutrophilic soil streptomycetes. Microbiology. 2004;73:74–8. https://doi.org/10.1023/b:mici.0000016372.52239.dd.

Ma A, Zhang X, Jiang K, Zhao C, Liu J, Wu M, Wang Y, Wang M, Li J, Xu S. Phylogenetic and physiological diversity of cultivable actinomycetes isolated from alpine habitats on the Qinghai-Tibetan Plateau. Front Microbiol. 2020;11: 555351. https://doi.org/10.3389/fmicb.2020.555351.

Romero-Rodríguez A, Maldonado-Carmona N, Ruiz-Villafán B, Koirala N, Rocha D, Sánchez S. Interplay between carbon, nitrogen and phosphate utilization in the control of secondary metabolite production in Streptomyces. Antonie Van Leeuwenhoek. 2018;111:761–81. https://doi.org/10.1007/s10482-018-1073-1.

Lee CC, Houghton P. Cytotoxicity of plants from Malaysia and Thailand used traditionally to treat cancer. J Ethnopharmacol. 2005;100:237–43. https://doi.org/10.1016/j.jep.2005.01.064.

Katoch M, Singh G, Sharma S, Gupta N, Sangwan PL, Saxena AK. Cytotoxic and antimicrobial activities of endophytic fungi isolated from Bacopa monnieri (L.) Pennell (Scrophulariaceae). BMC Complement Altern Med. 2014;14:52–60. https://doi.org/10.1186/1472-6882-14-52.

Yagi R, Doi M. Isolation of an antioxidative substance produced by Aspergillus repens. Biosci Biotechnol Biochem. 1999;63:932–3.

Greco G, Turrini E, Catanzaro E, Fimognari C. Marine anthraquinones: pharmacological and toxicological issues. Mar Drugs. 2021;19:272. https://doi.org/10.3390/md19050272.

Bovio E, Fauchon M, Toueix Y, Mehiri M, Varese GC, Hellio C. The sponge-associated fungus Eurotium chevalieri MUT 2316 and its bioactive molecules: potential applications in the field of antifouling. Mar Biotechnol. 2019;21:743–52. https://doi.org/10.1007/s10126-019-09920-y.

Kim K-S, Cui X, Lee D-S, Sohn JH, Yim JH, Kim Y-C, Oh H. Anti-inflammatory effect of neoechinulin a from the marine fungus Eurotium sp. SF-5989 through the suppression of NF-кB and p38 MAPK pathways in lipopolysaccharide-stimulated RAW2647 macrophages. Molecules. 2013;18:13245–59. https://doi.org/10.3390/molecules181113245.

Zhang L, Song T, Wu J, Zhang S, Yin C, Huang F, Hang Y, Abbas N, Liu X, Zhang Y. Antibacterial and cytotoxic metabolites of termite-associated Streptomyces sp. BYF63. J Antibiot. 2020;73:766–71. https://doi.org/10.1038/s41429-020-0334-1.

Zhao PJ, Wang HX, Li GH, Li HD, Liu J, Shen YM. Secondary metabolites from endophytic Streptomyces sp. Lz531. Chem Biodivers. 2007;4:899–904. https://doi.org/10.1002/cbdv.200790078.

Lai Z, Yu J, Ling H, Song Y, Yuan J, Ju J, Tao Y, Huang H. Grincamycins I-K, cytotoxic angucycline glycosides derived from marine-derived actinomycete Streptomyces lusitanus SCSIO LR32. Planta Med. 2018;84:201–7. https://doi.org/10.1055/s-0043-119888.

Královcová E, Blumauerová M, Vaněk Z. Strain improvement in Streptomyces galilaeus, a producer of anthracycline antibiotics galirubins. Folia Microbiol. 1977;22:321–8.

Harmeet M, Gores GJ, Lemasters JJ. Apoptosis and necrosis in the liver: a tale of two deaths? Hepatology. 2006;43:S31–44. https://doi.org/10.1002/hep.21062.

Chen Q, Kang J, Fu C. The independence of and associations among apoptosis, autophagy, and necrosis. Signal Transduct Target Ther. 2018;3:18–29. https://doi.org/10.1038/s41392-018-0018-5.

Schultz DR, Harringto Jr WJ. Apoptosis: programmed cell death at a molecular level. Semin Arthritis Rheum. 2003;32:345–69. https://doi.org/10.1053/sarh.2003.50005.

Susan E. A review of programmed cell death. Toxicol Pathol. 2007;35:495–516. https://doi.org/10.1080/01926230701320337.

Ghobrial IM, Witzig TE, Adjei AA. Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin. 2005;55:178–94. https://doi.org/10.3322/canjclin.55.3.178.

Kar S. Unraveling cell-cycle dynamics in cancer. Cell Syst. 2016;2:8–10. https://doi.org/10.1016/j.cels.2016.01.007.

Schwartz GK, Shah MA. Targeting the cell cycle: a new approach to cancer therapy. J Clin Oncol. 2005;23:9408–21. https://doi.org/10.1200/jco.2005.01.5594.

Das R, Romi W, Das R, Sharma HK, Thakur D. Antimicrobial potentiality of actinobacteria isolated from two microbiologically unexplored forest ecosystems of Northeast India. BMC Microbiol. 2018;18:71–71. https://doi.org/10.1186/s12866-018-1215-7.

Kumar V, Naik B, Gusain O, Bisht GS. An actinomycete isolate from solitary wasp mud nest having strong antibacterial activity and kills the Candida cells due to the shrinkage and the cytosolic loss. Front Microbiol. 2014;5:446–446. https://doi.org/10.3389/fmicb.2014.00446.

Brenner DJ, Krieg NR, Staley JT. Bergey’s manual® of systematic bacteriology. Baltimore: Williams & Wilkins; 2005.

Shirling EB, Gottlieb D. Methods for characterization of Streptomyces species. Int J Syst Bacteriol. 1966;16:313–40. https://doi.org/10.1099/00207713-16-3-313.

Orsini M, Romano-Spica V. A microwave-based method for nucleic acid isolation from environmental samples. Lett Appl Microbiol. 2001;33:17–20. https://doi.org/10.1046/j.1472-765x.2001.00938.x.

Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 1985;39:783–91. https://doi.org/10.1111/j.1558-5646.1985.tb00420.x.

Tian R, Li Y, Gao M. Shikonin causes cell-cycle arrest and induces apoptosis by regulating the EGFR-NF-κB signalling pathway in human epidermoid carcinoma A431 cells. Biosci Rep. 2015;35:e00189. https://doi.org/10.1042/BSR20150002.

Thông tin
Thông tin xuất bản

Microbial Cell Factories

Thông tin tác giả