Skip to main content
Top
Published in: BMC Complementary Medicine and Therapies 1/2023

Open Access 01-12-2023 | Research

Cytotoxic, anti-proliferative, and apoptotic evaluation of Ramalina sinensis (Ascomycota, Lecanoromycetes), lichenized fungus on oral squamous cell carcinoma cell line; in-vitro study

Authors: Maryam Koopaie, Hanieh Karimi, Mohammad Sohrabi, Hooman Norouzi

Published in: BMC Complementary Medicine and Therapies | Issue 1/2023

Login to get access

Abstract

Background

Scientists and medical professionals are actively striving to improve the efficacy of treatment methods for oral squamous cell carcinoma (OSCC), the most frequently occurring cancer within the oral cavity, by exploring the potential of natural products. The active pharmacological compounds found in lichenized fungi have shown potential for aiding in cancer treatment. Recent research aims to evaluate the impact of the lichenized fungus Ramalina sinensis (R. sinensis) on the cell viability and apoptosis of OSCC cell lines, considering the anti-inflammatory and anti-cancer capabilities of lichens.

Methods

Ramalina sinensis (Ascomycota, Lecanoromycetes) was selected for investigation of its effects on a human oral squamous cell carcinoma cell line. Acetone and methanol extracts of R. sinensis on an OSCC cell line (KB cell line, NCBI Code: C152) were investigated. Viability was assessed by MTT assay analysis, and apoptotic cells were measured using flow cytometry analysis. Scratch assay was used to assess cell migration. The chemical composition and metabolic profiling of R. sinensis were investigated.

Results

The growth and multiplication of KB cells were observed to undergo a gradual but remarkable inhibition when exposed to various concentrations. Specifically, concentrations of 6.25, 12.5, 25, 50, 100, and 200 μg/mL exhibited a significant suppressive effect on the proliferation of KB cells. The inhibition of cell proliferation exhibited a statistically significant difference between the extracts obtained from acetone and methanol. Flow cytometry results show an increase in apoptosis of OSCC cells by acetone extract. R. sinensis exerted a concentration-dependent inhibitory effect on the migration of OSCC cells. The chemical composition of R. sinensis was investigated using liquid chromatography positive ion electrospray ionization tandem mass spectrometry (LC–ESI–MS/MS), and 33 compounds in the acetone and methanol extracts of R. sinensis were detected.

Conclusion

The findings provide evidence supporting the beneficial effects of R. sinensis extract on inducing apoptosis in OSCC cells and exerting anti-cancer properties.
Appendix
Available only for authorised users
Literature
1.
go back to reference Vucicevic Boras V, Fucic A, Virag M, Gabric D, Blivajs I, Tomasovic-Loncaric C, et al. Significance of stroma in biology of oral squamous cell carcinoma. Tumori J. 2018;104(1):9–14.CrossRef Vucicevic Boras V, Fucic A, Virag M, Gabric D, Blivajs I, Tomasovic-Loncaric C, et al. Significance of stroma in biology of oral squamous cell carcinoma. Tumori J. 2018;104(1):9–14.CrossRef
3.
go back to reference Nandini D, Rao RS, Hosmani J, Khan S, Patil S, Awan KH. Novel therapies in the management of oral cancer: an update. Dis Mon. 2020;66(12):101036.CrossRefPubMed Nandini D, Rao RS, Hosmani J, Khan S, Patil S, Awan KH. Novel therapies in the management of oral cancer: an update. Dis Mon. 2020;66(12):101036.CrossRefPubMed
5.
go back to reference Popovici V, Bucur L, Vochita G, Gherghel D, Mihai CT, Rambu D, et al. In Vitro Anticancer Activity and Oxidative Stress Biomarkers Status Determined by Usnea barbata (L.) FH Wigg. Dry Extracts. Antioxidants. 2021;10(7):1141.CrossRefPubMedPubMedCentral Popovici V, Bucur L, Vochita G, Gherghel D, Mihai CT, Rambu D, et al. In Vitro Anticancer Activity and Oxidative Stress Biomarkers Status Determined by Usnea barbata (L.) FH Wigg. Dry Extracts. Antioxidants. 2021;10(7):1141.CrossRefPubMedPubMedCentral
6.
go back to reference Tripathi AH, Negi N, Gahtori R, Kumari A, Joshi P, Tewari LM, et al. A review of anti-cancer and related properties of lichen-extracts and metabolites. Anticancer Agents Med Chem. 2022;22(1):115–42.PubMed Tripathi AH, Negi N, Gahtori R, Kumari A, Joshi P, Tewari LM, et al. A review of anti-cancer and related properties of lichen-extracts and metabolites. Anticancer Agents Med Chem. 2022;22(1):115–42.PubMed
7.
go back to reference Suh SS, Kim TK, Kim JE, Hong J-M, Nguyen TTT, Han SJ, et al. Anticancer activity of ramalin, a secondary metabolite from the antarctic lichen Ramalina terebrata, against colorectal cancer cells. Molecules. 2017;22(8):1361.CrossRefPubMedPubMedCentral Suh SS, Kim TK, Kim JE, Hong J-M, Nguyen TTT, Han SJ, et al. Anticancer activity of ramalin, a secondary metabolite from the antarctic lichen Ramalina terebrata, against colorectal cancer cells. Molecules. 2017;22(8):1361.CrossRefPubMedPubMedCentral
8.
go back to reference Dandapat M, Paul S. Secondary metabolites from lichen Usnea longissima and its pharmacological relevance. Pharmacognosy Res. 2019;11(2):103–9. Dandapat M, Paul S. Secondary metabolites from lichen Usnea longissima and its pharmacological relevance. Pharmacognosy Res. 2019;11(2):103–9.
9.
go back to reference Zhao Y, Wang M, Xu B. A comprehensive review on secondary metabolites and health-promoting effects of edible lichen. J Funct Foods. 2021;80:104283.CrossRef Zhao Y, Wang M, Xu B. A comprehensive review on secondary metabolites and health-promoting effects of edible lichen. J Funct Foods. 2021;80:104283.CrossRef
11.
go back to reference Oh SO, Wang XY, Wang LS, Liu PG, Hur JS. A note on the lichen genus Ramalina (Ramalinaceae, Ascomycota) in the Hengduan Mountains in China. Mycobiology. 2014;42(3):229–40.CrossRefPubMedPubMedCentral Oh SO, Wang XY, Wang LS, Liu PG, Hur JS. A note on the lichen genus Ramalina (Ramalinaceae, Ascomycota) in the Hengduan Mountains in China. Mycobiology. 2014;42(3):229–40.CrossRefPubMedPubMedCentral
12.
go back to reference Ristic S, Rankovic B, Stamenkovic S. Biopharmaceutical potential of two Ramalina lichens and their metabolites. Curr Pharm Biotechnol. 2016;17(7):651–8.CrossRefPubMed Ristic S, Rankovic B, Stamenkovic S. Biopharmaceutical potential of two Ramalina lichens and their metabolites. Curr Pharm Biotechnol. 2016;17(7):651–8.CrossRefPubMed
13.
go back to reference Shrestha G, St. Clair LL. Lichens: a promising source of antibiotic and anticancer drugs. Phytochem Rev. 2013;12(1):229–44.CrossRef Shrestha G, St. Clair LL. Lichens: a promising source of antibiotic and anticancer drugs. Phytochem Rev. 2013;12(1):229–44.CrossRef
14.
go back to reference Mohammadi M, Bagheri L, Badreldin A, Fatehi P, Pakzad L, Suntres Z, et al. Biological effects of gyrophoric acid and other lichen derived metabolites, on cell proliferation, apoptosis and cell signaling pathways. Chem Biol Interact. 2022;351:109768.CrossRefPubMed Mohammadi M, Bagheri L, Badreldin A, Fatehi P, Pakzad L, Suntres Z, et al. Biological effects of gyrophoric acid and other lichen derived metabolites, on cell proliferation, apoptosis and cell signaling pathways. Chem Biol Interact. 2022;351:109768.CrossRefPubMed
15.
go back to reference Bačkorová M, Jendželovský R, Kello M, Bačkor M, Mikeš J, Fedoročko P. Lichen secondary metabolites are responsible for induction of apoptosis in HT-29 and A2780 human cancer cell lines. Toxicol In Vitro. 2012;26(3):462–8.CrossRefPubMed Bačkorová M, Jendželovský R, Kello M, Bačkor M, Mikeš J, Fedoročko P. Lichen secondary metabolites are responsible for induction of apoptosis in HT-29 and A2780 human cancer cell lines. Toxicol In Vitro. 2012;26(3):462–8.CrossRefPubMed
16.
go back to reference Haraldsdóttir S, Guolaugsdóttir E, Ingólfsdóttir K, Ogmundsdóttir HM. Anti-proliferative effects of lichen-derived lipoxygenase inhibitors on twelve human cancer cell lines of different tissue origin in vitro. Planta Med. 2004;70(11):1098–100.CrossRefPubMed Haraldsdóttir S, Guolaugsdóttir E, Ingólfsdóttir K, Ogmundsdóttir HM. Anti-proliferative effects of lichen-derived lipoxygenase inhibitors on twelve human cancer cell lines of different tissue origin in vitro. Planta Med. 2004;70(11):1098–100.CrossRefPubMed
17.
18.
go back to reference Kumar J, Dhar P, Tayade AB, Gupta D, Chaurasia OP, Upreti DK, et al. Antioxidant capacities, phenolic profile and cytotoxic effects of saxicolous lichens from trans-Himalayan cold desert of Ladakh. PLoS One. 2014;9(6):e98696.CrossRefPubMedPubMedCentral Kumar J, Dhar P, Tayade AB, Gupta D, Chaurasia OP, Upreti DK, et al. Antioxidant capacities, phenolic profile and cytotoxic effects of saxicolous lichens from trans-Himalayan cold desert of Ladakh. PLoS One. 2014;9(6):e98696.CrossRefPubMedPubMedCentral
19.
go back to reference Akram SM, Al-Saffar AZ, Hadi NA, Akram SM. Utilization of novel lectin-conjugated Au nanoparticles as Thomsen-Friedenreich onco-antigen target for in vitro cytotoxicity and apoptosis induction in leukemic cell line. Life Sci. 2022;311:121163.CrossRefPubMed Akram SM, Al-Saffar AZ, Hadi NA, Akram SM. Utilization of novel lectin-conjugated Au nanoparticles as Thomsen-Friedenreich onco-antigen target for in vitro cytotoxicity and apoptosis induction in leukemic cell line. Life Sci. 2022;311:121163.CrossRefPubMed
20.
go back to reference Mahmood RI, Abbass AK, Al-Saffar AZ, Al-Obaidi JR. An in vitro cytotoxicity of a novel pH-Sensitive lectin loaded-cockle shell-derived calcium carbonate nanoparticles against MCF-7 breast tumour cell. J Drug Deliv Sci Technol. 2021;61:102230.CrossRef Mahmood RI, Abbass AK, Al-Saffar AZ, Al-Obaidi JR. An in vitro cytotoxicity of a novel pH-Sensitive lectin loaded-cockle shell-derived calcium carbonate nanoparticles against MCF-7 breast tumour cell. J Drug Deliv Sci Technol. 2021;61:102230.CrossRef
21.
go back to reference Mahmood RI, Abbass AK, Razali N, Al-Saffar AZ, Al-Obaidi JR. Protein profile of MCF-7 breast cancer cell line treated with lectin delivered by CaCO3NPs revealed changes in molecular chaperones, cytoskeleton, and membrane-associated proteins. Int J Biol Macromol. 2021;184:636–47.CrossRefPubMed Mahmood RI, Abbass AK, Razali N, Al-Saffar AZ, Al-Obaidi JR. Protein profile of MCF-7 breast cancer cell line treated with lectin delivered by CaCO3NPs revealed changes in molecular chaperones, cytoskeleton, and membrane-associated proteins. Int J Biol Macromol. 2021;184:636–47.CrossRefPubMed
22.
go back to reference Norouzi H, Azizi A, Gholami M, Sohrabi M, Boustie J. Chemotype variations among lichen ecotypes of Umbilicaria aprina as revealed by LC-ESI-MS/MS: a survey of antioxidant phenolics. Environ Sci Pollut Res. 2020;27(32):40296–308.CrossRef Norouzi H, Azizi A, Gholami M, Sohrabi M, Boustie J. Chemotype variations among lichen ecotypes of Umbilicaria aprina as revealed by LC-ESI-MS/MS: a survey of antioxidant phenolics. Environ Sci Pollut Res. 2020;27(32):40296–308.CrossRef
23.
go back to reference Olivier-Jimenez D, Chollet-Krugler M, Rondeau D, Beniddir MA, Ferron S, Delhaye T, et al. A database of high-resolution MS/MS spectra for lichen metabolites. Sci Data. 2019;6(1):1–11.CrossRef Olivier-Jimenez D, Chollet-Krugler M, Rondeau D, Beniddir MA, Ferron S, Delhaye T, et al. A database of high-resolution MS/MS spectra for lichen metabolites. Sci Data. 2019;6(1):1–11.CrossRef
24.
go back to reference Nazari J, Payamnoor V, Kavosi M, Asadi J. Extraction of anti-cancer triterpenoids (betulinic acid and betulin) from the birch bark-inhabiting lichen (Ramalina sinensis). Iran J Med Aromat Plants Res. 2018;34(4):604–16. Nazari J, Payamnoor V, Kavosi M, Asadi J. Extraction of anti-cancer triterpenoids (betulinic acid and betulin) from the birch bark-inhabiting lichen (Ramalina sinensis). Iran J Med Aromat Plants Res. 2018;34(4):604–16.
25.
go back to reference Lee E, Lee CG, Yim JH, Lee HK, Pyo S. Ramalin-mediated apoptosis is enhanced by autophagy inhibition in human breast cancer cells. Phytother Res. 2016;30(3):426–38.CrossRefPubMed Lee E, Lee CG, Yim JH, Lee HK, Pyo S. Ramalin-mediated apoptosis is enhanced by autophagy inhibition in human breast cancer cells. Phytother Res. 2016;30(3):426–38.CrossRefPubMed
26.
27.
go back to reference Nguyen TT, Yoon S, Yang Y, Lee H-B, Oh S, Jeong M-H, et al. Lichen secondary metabolites in Flavocetraria cucullata exhibit anti-cancer effects on human cancer cells through the induction of apoptosis and suppression of tumorigenic potentials. PLoS One. 2014;9(10):e111575.CrossRefPubMedPubMedCentral Nguyen TT, Yoon S, Yang Y, Lee H-B, Oh S, Jeong M-H, et al. Lichen secondary metabolites in Flavocetraria cucullata exhibit anti-cancer effects on human cancer cells through the induction of apoptosis and suppression of tumorigenic potentials. PLoS One. 2014;9(10):e111575.CrossRefPubMedPubMedCentral
28.
go back to reference Aoussar N, Laasri FE, Bourhia M, Manoljovic N, Mhand RA, Rhallabi N, et al. Phytochemical analysis, cytotoxic, antioxidant, and antibacterial activities of lichens. Evid Based Complement Alternat Med. 2020;2020:8104538. Aoussar N, Laasri FE, Bourhia M, Manoljovic N, Mhand RA, Rhallabi N, et al. Phytochemical analysis, cytotoxic, antioxidant, and antibacterial activities of lichens. Evid Based Complement Alternat Med. 2020;2020:8104538.
29.
go back to reference Yang Y, Bae WK, Lee J-Y, Choi YJ, Lee KH, Park M-S, et al. Potassium usnate, a water-soluble usnic acid salt, shows enhanced bioavailability and inhibits invasion and metastasis in colorectal cancer. Sci Rep. 2018;8(1):16234.CrossRefPubMedPubMedCentral Yang Y, Bae WK, Lee J-Y, Choi YJ, Lee KH, Park M-S, et al. Potassium usnate, a water-soluble usnic acid salt, shows enhanced bioavailability and inhibits invasion and metastasis in colorectal cancer. Sci Rep. 2018;8(1):16234.CrossRefPubMedPubMedCentral
30.
go back to reference Xu M, Heidmarsson S, Olafsdottir ES, Buonfiglio R, Kogej T, Omarsdottir S. Secondary metabolites from cetrarioid lichens: Chemotaxonomy, biological activities and pharmaceutical potential. Phytomedicine. 2016;23(5):441–59.CrossRefPubMed Xu M, Heidmarsson S, Olafsdottir ES, Buonfiglio R, Kogej T, Omarsdottir S. Secondary metabolites from cetrarioid lichens: Chemotaxonomy, biological activities and pharmaceutical potential. Phytomedicine. 2016;23(5):441–59.CrossRefPubMed
31.
go back to reference Yang Y, Bhosle SR, Yu YH, Park S-Y, Zhou R, Taş İ, et al. Tumidulin, a lichen secondary metabolite, decreases the stemness potential of colorectal cancer cells. Molecules. 2018;23(11):2968.CrossRefPubMedPubMedCentral Yang Y, Bhosle SR, Yu YH, Park S-Y, Zhou R, Taş İ, et al. Tumidulin, a lichen secondary metabolite, decreases the stemness potential of colorectal cancer cells. Molecules. 2018;23(11):2968.CrossRefPubMedPubMedCentral
32.
go back to reference Cardile V, Graziano A, Avola R, Piovano M, Russo A. Potential anticancer activity of lichen secondary metabolite physodic acid. Chem Biol Interact. 2017;263:36–45.CrossRefPubMed Cardile V, Graziano A, Avola R, Piovano M, Russo A. Potential anticancer activity of lichen secondary metabolite physodic acid. Chem Biol Interact. 2017;263:36–45.CrossRefPubMed
33.
go back to reference Ulus G. Antiangiogenic properties of lichen secondary metabolites. Phytother Res. 2021;35(6):3046–58.CrossRefPubMed Ulus G. Antiangiogenic properties of lichen secondary metabolites. Phytother Res. 2021;35(6):3046–58.CrossRefPubMed
Metadata
Title
Cytotoxic, anti-proliferative, and apoptotic evaluation of Ramalina sinensis (Ascomycota, Lecanoromycetes), lichenized fungus on oral squamous cell carcinoma cell line; in-vitro study
Authors
Maryam Koopaie
Hanieh Karimi
Mohammad Sohrabi
Hooman Norouzi
Publication date
01-12-2023
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2023
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-023-04118-1

Other articles of this Issue 1/2023

BMC Complementary Medicine and Therapies 1/2023 Go to the issue