Comparison of cytotoxicity caused by Viscum album in human mesenchymal stem cells and hepatocellular carcinoma cells
Main Article Content
Abstract
Viscum album (VA), also known as Mistletoe, has various therapeutic properties, including analgesic, anti-inflammatory, and anticancer effects. It has been used for treating different types of cancer, exhibiting proven efficacy against breast cancer, glioblastoma, carcinoma, and other advanced-stage tumor types. Besides promoting improvements in the clinical condition, VA also helps reduce the side effects caused by conventional treatment, thereby offering patients a better quality of life. Hence, the objective of this study was to assess the effects of the homeopathic dilution of VA at the potency of D30 (1x10-30) (VA D30) on human mesenchymal stem cells (MSCs) and hepatocellular carcinoma cells (HepG2). The cells were grown in 75 cm² flasks until they reached approximately 80% confluence. Subsequently, they were trypsinized and plated in 96-well plates at 10,000 cells/well. After 24 hours of incubation in an oven, VA was added at 30 and 40 µL/mL concentrations. The cells were further incubated for an additional 48 hours. After the treatment, the cells underwent a cell viability test using MTT. The results indicated a decrease in HepG2 viability, while no damage to normal cells (MSCs) was detected. The findings suggest that VAD30 holds promise as a potential therapeutic agent in treating hepatocellular carcinoma due to its observed cytotoxicity towards HepG2 cells while exhibiting no adverse effects on mesenchymal stem cells at the equivalent concentration.
Keywords:
cytotoxicity, stem cells, hepatocellular carcinoma cells
Article Details
How to Cite
VALLE, Ana Catarina Viana et al.
Comparison of cytotoxicity caused by Viscum album in human mesenchymal stem cells and hepatocellular carcinoma cells.
Medical Research Archives, [S.l.], v. 12, n. 5, may 2024.
ISSN 2375-1924.
Available at: <https://esmed.org/MRA/mra/article/view/5136>. Date accessed: 30 june 2024.
doi: https://doi.org/10.18103/mra.v12i5.5136.
Section
Research Articles
The Medical Research Archives grants authors the right to publish and reproduce the unrevised contribution in whole or in part at any time and in any form for any scholarly non-commercial purpose with the condition that all publications of the contribution include a full citation to the journal as published by the Medical Research Archives.
References
1. Szurpnicka A, Kowalczuk A, Szterk A. Biological activity of Mistletoe: in vitro and in vivo studies and mechanisms of action. Arch Pharm Res. 2020;43(6):593-629. doi:10.1007/ s12272-020-01247-w
2. Nicoletti M. The Anti-Inflammatory Activity of Viscum album. Plants (Basel). 2023;12(7):14 60. Published 2023 Mar 27. doi:10.3390/plant s12071460
3. Zänker KS, Kaveri SV, editors. Mistletoe: From Mythology to Evidence-Based Medicine. Karger; Basel, Switzerland: 2015; 4:1–10. Translational Research in Biomedicine. doi: 10.4081/ejh.2015.2577
4. Melo MNO, Ochioni AC, Zancan P, et al. Viscum album mother tinctures: Harvest conditions and host trees influence the plant metabolome and the glycolytic pathway of breast cancer cells. Front Pharmacol. 2022; 13:1027931. Published 2022 Oct 31. doi:10.3 389/fphar.2022.1027931
5. Schötterl S, Miemietz JT, Ilina EI, et al. Mistletoe-Based Drugs Work in Synergy with Radio-Chemotherapy in the Treatment of Glioma In Vitro and In Vivo in Glioblastoma Bearing Mice. Evid Based Complement Alternat Med. 2019; 2019:1376140. Published 2019 Jul 3. doi:10.1155/2019/1376140
6. Reynel M, Villegas Y, Kiene H, Werthmann PG, Kienle GS. Intralesional and subcutaneous application of Viscum album L. (European mistletoe) extract in cervical carcinoma in situ: A CARE compliant case report [published correction appears in Medicine (Baltimore). 2018 Dec;97(52):e13950]. Medicine (Baltimore). 2018;97(48):e13420. doi:10.1097/MD.0000000000013420
7. Schad F, Thronicke A, Steele ML, et al. Overall survival of stage IV non-small cell lung cancer patients treated with Viscum album L. in addition to chemotherapy, a real-world observational multicenter analysis [published correction appears in PLoS One. 2022 Aug 16;17(8): e0273387]. PLoS One. 2018;13(8):e0 203058. Published 2018 Aug 27. doi:10.1371/ journal.pone.0203058
8. Kissel D, Jurin M, Zarcovic N. Über die zytostatischen und immunologischen Effekte von Viscum album. Erfahrunsheilkunde 1990;39:973–988
9. Gabius HJ, Gabius S, Joshi SS, et al. From ill-defined extracts to the immunomodulatory lectin: will there be a reason for oncological application of Mistletoe? Planta Med 1994;60: 2–7. doi: 10.1055/s-2006-959396
10. Szurpnicka A, Zjawiony JK, Szterk A. Therapeutic potential of Mistletoe in CNS-related neurological disorders and the chemical composition of Viscum species. J Ethnopharmacol. 2019; 231:241-252. doi:10. 1016/j.jep.2018.11.025
11. Khil LY, Kim W, Lyu S, Park WB, Yoon JW, Jun HS. Mechanisms involved in Korean Mistletoe lectin-induced apoptosis of cancer cells. World J Gastroenterol 2007;13:2811–2818. doi: 10.3748/wjg.v13.i20.2811
12. Lee C-H, Kim J-K, Kim H-Y, Park S-M, Lee S-M. Immunomodulating effects of Korean mistletoe lectin in vitro and in vivo. Int Immunopharmacol 2009;9:1555–1561. doi: 10.1016/j.intimp.2009.09.011
13. Nazaruk J, Orlikowski P. Phytochemical profile and therapeutic potential of Viscum album L. Nat Prod Res 2016;30:373–385. doi: 10.1080/14786419.2015.1022776
14. Delebinski CI, Twardziok M, Kleinsimon S, et al. A natural combination extract of Viscum album L. Containing both triterpene acids and lectins is highly effective against AML in vivo. PLoS One 2015;10:e0133892. doi: 10.1371/jo urnal.pone.0133892
15. Loef M, Walach H. Quality of life in cancer patients treated with Mistletoe: a systematic review and meta-analysis. BMC Complement Med Ther. 2020;20(1):227. Published 2020 Jul 20. doi:10.1186/s12906-020-03013-3
16. Park YK, Do YR, Jang BC. Apoptosis of K562 leukemia cells by Abnobaviscum F®, a European mistletoe extract. Oncol Rep. 2012;28(6):2227-2232. doi:10.3892/or.2012.2026
17. Fan J, Wu M, Wang J, Ren D, Zhao J, Yang G. 1,7-Bis(4-hydroxyphenyl)-1,4-heptadien-3-one induces lung cancer cell apoptosis via the PI3K/Akt and ERK1/2 pathways. J Cell Physiol. 2019;234(5):6336-6349. doi:10.1002/jcp.27364
18. dela Cruz JF, Kim YS, Lumbera WM, Hwang SG. Viscum Album Var Hot Water Extract Mediates Anticancer Effects through G1 Phase Cell Cycle Arrest in SK-Hep1 Human Hepatocarcinoma cells. Asian Pac J Cancer Prev. 2015;16(15):6417-6421. doi:10.7314/ap jcp.2015.16.15.6417
19. Mishra R, Sharma S, Sharma RS, et al. Viscum articulatum Burm. f. aqueous extract exerts antiproliferative effect and induces cell cycle arrest and apoptosis in leukemia cells. J Ethnopharmacol. 2018;219:91-102. doi:10.10 16/j.jep.2018.03.005
20. Twardziok M, Kleinsimon S, Rolff J, et al. Multiple Active Compounds from Viscum album L. Synergistically Converge to Promote Apoptosis in Ewing Sarcoma. PLoS One. 2016;11(9):e0159749. Published 2016 Sep 2. doi:10.1371/journal.pone.0159749
21. Hegazy MG, Imam AM, Abdelghany BE. Evaluation of cytotoxic and anticancer effect of Orobanche crenata methanolic extract on cancer cell lines. Tumor Biol 2020;42:1-11. doi: 10.1177/1010428320918685
22. Siegel RL, Miller KD, Wagle NS, Jemal A. Cancer statistics, 2023. CA Cancer J Clin. 2023;73(1):17-48. doi:10.3322/caac.21763
23. Chidambaranathan-Reghupaty S, Fisher PB, Sarkar D. Hepatocellular carcinoma (HCC): Epidemiology, etiology and molecular classification. Adv Cancer Res. 2021;149: 1-61. doi:10.1016/bs.acr.2020.10.001
24. Marvibaigi M, Supriyanto E, Amini N, Abdul Majid FA, Jaganathan SK. Preclinical and clinical effects of Mistletoe against breast cancer. BioMed Res Int 2014;2014:785479. doi: 10.1155/2014/785479
25. Estko M, Baumgartner S, Urech K, et al. Tumour cell derived effects on monocyte/ macrophage polarization and function and modulatory potential of Viscum album lipophilic extract in vitro. BMC Complement Altern Med 2015;15:130. doi: 10.1186/s1290 6-015-0650-3
26. Laszczyk MN. Pentacyclic triterpenes of the lupane, oleanane and ursane group as tools in cancer therapy. Planta Med 2009; 75:1549–1560. doi: 10.1055/s-0029-1186102
27. Strüh CM, Jäger S, Kersten A, Schempp CM, Scheffler A, Martin SF. Triterpenoids amplify anti-tumoral effects of mistletoe extracts on murine B16.f10 melanoma in vivo. PLoS One 2013;8:e62168. doi: 10.1371/journ al.pone.0062168
28. Abud APR, Zych J, Reus TL, et al. The use of human adipose-derived stem cells based cytotoxicity assay for acute toxicity test. Regul Toxicol Pharmacol 2015;73:992–998. doi: 10.1016/j.yrtph.2015.09.015
29. Valle ACV, Brunel HDSS, Dallago BSL, et al. In-Vitro Growth Kinetics of Mesenchymal Stem Cells in Cytotoxicity Tests Using Low-Diluted Viscum Album. Homeopathy. 2023;11 2(1):40-49. doi:10.1055/s-0042-1747682
30. Steinborn C, Klemd AM, Sanchez-Campillo AS, et al. Viscum album neutralizes tumor-induced immunosuppression in a human in vitro cell model [published correction appears in PLoS One. 2017 Nov 2;12 (11): e0187786]. PLoS One. 2017;12(7):e 0181553. Published 2017 Jul 18. doi:10.1371/ journal.pone.0181553
31. Pascale RM, Calvisi DF, Simile MM, Feo CF, Feo F. The Warburg Effect 97 Years after Its Discovery. Cancers (Basel). 2020;12(10):28 19. Published 2020 Sep 30. doi:10.3390/canc ers12102819
32. Melo MNO, Ochioni AC, Zancan P, et al. Viscum album mother tinctures: Harvest conditions and host trees influence the plant metabolome and the glycolytic pathway of breast cancer cells. Front Pharmacol. 2022;13: 1027931. Published 2022 Oct 31. doi:10.3389 /fphar.2022.1027931
33. Mavrikou, S, Tsekouras, V, Karageorgou, MA et al. Anticancer and biochemical effects of Viscum album L. protein extracts on HeLa cells. Plant Cell Tiss Organ Cult 140, 369–378 (2020). doi: 10.1007/s11240-019-01733-0
34. Park YR, Jee W, Park SM, et al. Viscum album Induces Apoptosis by Regulating STAT3 Signaling Pathway in Breast Cancer Cells. Int J Mol Sci. 2023; 24(15):11988. Published 2023 Jul 26. doi:10.3390/ijms2415 11988
35. Lee C, Cheung ST. STAT3: An Emerging Therapeutic Target for Hepatocellular Carcinoma. Cancers (Basel). 2019;11(11):1646 . Published 2019 Oct 25. doi:10.3390/cancers 11111646
2. Nicoletti M. The Anti-Inflammatory Activity of Viscum album. Plants (Basel). 2023;12(7):14 60. Published 2023 Mar 27. doi:10.3390/plant s12071460
3. Zänker KS, Kaveri SV, editors. Mistletoe: From Mythology to Evidence-Based Medicine. Karger; Basel, Switzerland: 2015; 4:1–10. Translational Research in Biomedicine. doi: 10.4081/ejh.2015.2577
4. Melo MNO, Ochioni AC, Zancan P, et al. Viscum album mother tinctures: Harvest conditions and host trees influence the plant metabolome and the glycolytic pathway of breast cancer cells. Front Pharmacol. 2022; 13:1027931. Published 2022 Oct 31. doi:10.3 389/fphar.2022.1027931
5. Schötterl S, Miemietz JT, Ilina EI, et al. Mistletoe-Based Drugs Work in Synergy with Radio-Chemotherapy in the Treatment of Glioma In Vitro and In Vivo in Glioblastoma Bearing Mice. Evid Based Complement Alternat Med. 2019; 2019:1376140. Published 2019 Jul 3. doi:10.1155/2019/1376140
6. Reynel M, Villegas Y, Kiene H, Werthmann PG, Kienle GS. Intralesional and subcutaneous application of Viscum album L. (European mistletoe) extract in cervical carcinoma in situ: A CARE compliant case report [published correction appears in Medicine (Baltimore). 2018 Dec;97(52):e13950]. Medicine (Baltimore). 2018;97(48):e13420. doi:10.1097/MD.0000000000013420
7. Schad F, Thronicke A, Steele ML, et al. Overall survival of stage IV non-small cell lung cancer patients treated with Viscum album L. in addition to chemotherapy, a real-world observational multicenter analysis [published correction appears in PLoS One. 2022 Aug 16;17(8): e0273387]. PLoS One. 2018;13(8):e0 203058. Published 2018 Aug 27. doi:10.1371/ journal.pone.0203058
8. Kissel D, Jurin M, Zarcovic N. Über die zytostatischen und immunologischen Effekte von Viscum album. Erfahrunsheilkunde 1990;39:973–988
9. Gabius HJ, Gabius S, Joshi SS, et al. From ill-defined extracts to the immunomodulatory lectin: will there be a reason for oncological application of Mistletoe? Planta Med 1994;60: 2–7. doi: 10.1055/s-2006-959396
10. Szurpnicka A, Zjawiony JK, Szterk A. Therapeutic potential of Mistletoe in CNS-related neurological disorders and the chemical composition of Viscum species. J Ethnopharmacol. 2019; 231:241-252. doi:10. 1016/j.jep.2018.11.025
11. Khil LY, Kim W, Lyu S, Park WB, Yoon JW, Jun HS. Mechanisms involved in Korean Mistletoe lectin-induced apoptosis of cancer cells. World J Gastroenterol 2007;13:2811–2818. doi: 10.3748/wjg.v13.i20.2811
12. Lee C-H, Kim J-K, Kim H-Y, Park S-M, Lee S-M. Immunomodulating effects of Korean mistletoe lectin in vitro and in vivo. Int Immunopharmacol 2009;9:1555–1561. doi: 10.1016/j.intimp.2009.09.011
13. Nazaruk J, Orlikowski P. Phytochemical profile and therapeutic potential of Viscum album L. Nat Prod Res 2016;30:373–385. doi: 10.1080/14786419.2015.1022776
14. Delebinski CI, Twardziok M, Kleinsimon S, et al. A natural combination extract of Viscum album L. Containing both triterpene acids and lectins is highly effective against AML in vivo. PLoS One 2015;10:e0133892. doi: 10.1371/jo urnal.pone.0133892
15. Loef M, Walach H. Quality of life in cancer patients treated with Mistletoe: a systematic review and meta-analysis. BMC Complement Med Ther. 2020;20(1):227. Published 2020 Jul 20. doi:10.1186/s12906-020-03013-3
16. Park YK, Do YR, Jang BC. Apoptosis of K562 leukemia cells by Abnobaviscum F®, a European mistletoe extract. Oncol Rep. 2012;28(6):2227-2232. doi:10.3892/or.2012.2026
17. Fan J, Wu M, Wang J, Ren D, Zhao J, Yang G. 1,7-Bis(4-hydroxyphenyl)-1,4-heptadien-3-one induces lung cancer cell apoptosis via the PI3K/Akt and ERK1/2 pathways. J Cell Physiol. 2019;234(5):6336-6349. doi:10.1002/jcp.27364
18. dela Cruz JF, Kim YS, Lumbera WM, Hwang SG. Viscum Album Var Hot Water Extract Mediates Anticancer Effects through G1 Phase Cell Cycle Arrest in SK-Hep1 Human Hepatocarcinoma cells. Asian Pac J Cancer Prev. 2015;16(15):6417-6421. doi:10.7314/ap jcp.2015.16.15.6417
19. Mishra R, Sharma S, Sharma RS, et al. Viscum articulatum Burm. f. aqueous extract exerts antiproliferative effect and induces cell cycle arrest and apoptosis in leukemia cells. J Ethnopharmacol. 2018;219:91-102. doi:10.10 16/j.jep.2018.03.005
20. Twardziok M, Kleinsimon S, Rolff J, et al. Multiple Active Compounds from Viscum album L. Synergistically Converge to Promote Apoptosis in Ewing Sarcoma. PLoS One. 2016;11(9):e0159749. Published 2016 Sep 2. doi:10.1371/journal.pone.0159749
21. Hegazy MG, Imam AM, Abdelghany BE. Evaluation of cytotoxic and anticancer effect of Orobanche crenata methanolic extract on cancer cell lines. Tumor Biol 2020;42:1-11. doi: 10.1177/1010428320918685
22. Siegel RL, Miller KD, Wagle NS, Jemal A. Cancer statistics, 2023. CA Cancer J Clin. 2023;73(1):17-48. doi:10.3322/caac.21763
23. Chidambaranathan-Reghupaty S, Fisher PB, Sarkar D. Hepatocellular carcinoma (HCC): Epidemiology, etiology and molecular classification. Adv Cancer Res. 2021;149: 1-61. doi:10.1016/bs.acr.2020.10.001
24. Marvibaigi M, Supriyanto E, Amini N, Abdul Majid FA, Jaganathan SK. Preclinical and clinical effects of Mistletoe against breast cancer. BioMed Res Int 2014;2014:785479. doi: 10.1155/2014/785479
25. Estko M, Baumgartner S, Urech K, et al. Tumour cell derived effects on monocyte/ macrophage polarization and function and modulatory potential of Viscum album lipophilic extract in vitro. BMC Complement Altern Med 2015;15:130. doi: 10.1186/s1290 6-015-0650-3
26. Laszczyk MN. Pentacyclic triterpenes of the lupane, oleanane and ursane group as tools in cancer therapy. Planta Med 2009; 75:1549–1560. doi: 10.1055/s-0029-1186102
27. Strüh CM, Jäger S, Kersten A, Schempp CM, Scheffler A, Martin SF. Triterpenoids amplify anti-tumoral effects of mistletoe extracts on murine B16.f10 melanoma in vivo. PLoS One 2013;8:e62168. doi: 10.1371/journ al.pone.0062168
28. Abud APR, Zych J, Reus TL, et al. The use of human adipose-derived stem cells based cytotoxicity assay for acute toxicity test. Regul Toxicol Pharmacol 2015;73:992–998. doi: 10.1016/j.yrtph.2015.09.015
29. Valle ACV, Brunel HDSS, Dallago BSL, et al. In-Vitro Growth Kinetics of Mesenchymal Stem Cells in Cytotoxicity Tests Using Low-Diluted Viscum Album. Homeopathy. 2023;11 2(1):40-49. doi:10.1055/s-0042-1747682
30. Steinborn C, Klemd AM, Sanchez-Campillo AS, et al. Viscum album neutralizes tumor-induced immunosuppression in a human in vitro cell model [published correction appears in PLoS One. 2017 Nov 2;12 (11): e0187786]. PLoS One. 2017;12(7):e 0181553. Published 2017 Jul 18. doi:10.1371/ journal.pone.0181553
31. Pascale RM, Calvisi DF, Simile MM, Feo CF, Feo F. The Warburg Effect 97 Years after Its Discovery. Cancers (Basel). 2020;12(10):28 19. Published 2020 Sep 30. doi:10.3390/canc ers12102819
32. Melo MNO, Ochioni AC, Zancan P, et al. Viscum album mother tinctures: Harvest conditions and host trees influence the plant metabolome and the glycolytic pathway of breast cancer cells. Front Pharmacol. 2022;13: 1027931. Published 2022 Oct 31. doi:10.3389 /fphar.2022.1027931
33. Mavrikou, S, Tsekouras, V, Karageorgou, MA et al. Anticancer and biochemical effects of Viscum album L. protein extracts on HeLa cells. Plant Cell Tiss Organ Cult 140, 369–378 (2020). doi: 10.1007/s11240-019-01733-0
34. Park YR, Jee W, Park SM, et al. Viscum album Induces Apoptosis by Regulating STAT3 Signaling Pathway in Breast Cancer Cells. Int J Mol Sci. 2023; 24(15):11988. Published 2023 Jul 26. doi:10.3390/ijms2415 11988
35. Lee C, Cheung ST. STAT3: An Emerging Therapeutic Target for Hepatocellular Carcinoma. Cancers (Basel). 2019;11(11):1646 . Published 2019 Oct 25. doi:10.3390/cancers 11111646