Top

BMC Complementary Medicine and Therapies

Published in:

Open Access 01-12-2021 | Metastasis | Research article

Vincetoxicum arnottianum modulates motility features and metastatic marker expression in pediatric rhabdomyosarcoma by stabilizing the actin cytoskeleton

Authors: Anna Adamus, Iftikhar Ali, Vasileios Vasileiadis, Luai Al-Hileh, Jan Lisec, Marcus Frank, Guido Seitz, Nadja Engel

Published in: BMC Complementary Medicine and Therapies | Issue 1/2021

Abstract

Background

Prevention of metastatic invasion is one of the main challenges in the treatment of alveolar rhabdomyosarcoma. Still the therapeutic options are limited. Therefore, an anti-tumor screening was initiated focusing on the anti-metastatic and anti-invasion properties of selected medicinal plant extracts and phytoestrogens, already known to be effective in the prevention and treatment of different cancer entities.

Methods

Treatment effects were first evaluated by cell viability, migration, invasion, and colony forming assays on the alveolar rhabdomyosarcoma cell line RH-30 in comparison with healthy primary cells.

Results

Initial anti-tumor screenings of all substances analyzed in this study, identified the plant extract of Vincetoxicum arnottianum (VSM) as the most promising candidate, harboring the highest anti-metastatic potential. Those significant anti-motility properties were proven by a reduced ability for migration (60%), invasion (99%) and colony formation (61%) under 48 h exposure to 25 μg/ml VSM. The restricted motility features were due to an induction of the stabilization of the cytoskeleton – actin fibers were 2.5-fold longer and were spanning the entire cell. Decreased proliferation (PCNA, AMT, GCSH) and altered metastasis (e. g. SGPL1, CXCR4, stathmin) marker expression on transcript and protein level confirmed the significant lowered tumorigenicity under VSM treatment. Finally, significant alterations in the cell metabolism were detected for 25 metabolites, with levels of uracil, N-acetyl serine and propanoyl phosphate harboring the greatest alterations. Compared to the conventional therapy with cisplatin, VSM treated cells demonstrated a similar metabolic shutdown of the primary cell metabolism. Primary control cells were not affected by the VSM treatment.

Conclusions

This study revealed the VSM root extract as a potential, new migrastatic drug candidate for the putative treatment of pediatric alveolar rhabdomyosarcoma with actin filament stabilizing properties and accompanied by a marginal effect on the vitality of primary cells.

Graphical abstract

Available only for authorised users

Chen C, Dorado GH, Scheer M, Henssen AG. Current and future treatment strategies for Rhabdomyosarcoma. Front Oncol. 2019;9:1458. https://doi.org/10.3389/fonc.2019.01458 .CrossRefPubMedPubMedCentral

Pal A, Chiu HY, Taneja R. Genetics, epigenetics and redox homeostasis in rhabdomyosarcoma: emerging targets and therapeutics. Redox Biol. 2019;25:101124. https://doi.org/10.1016/j.redox.2019.101124 .CrossRefPubMedPubMedCentral

Xie Z, Babiceanu M, Kumar S, Jia Y, Qin F, Barr FG, et al. Fusion transcriptome profiling provides insights into alveolar rhabdomyosarcoma. Proc Natl Acad Sci U S A. 2016;113(46):13126–31. https://doi.org/10.1073/pnas.1612734113 .CrossRefPubMedPubMedCentral

Shern JF, Yohe ME, Khan J. Pediatric Rhabdomyosarcoma. Crit Rev Oncog. 2015;20(3-4):227–43. https://doi.org/10.1615/CritRevOncog.2015013800 .CrossRefPubMedPubMedCentral

Rudzinski ER, Anderson JR, Hawkins DS, Skapek SX, Parham DM, Teot LA. The World Health Organization classification of skeletal muscle tumors in pediatric Rhabdomyosarcoma: a report from the Children’s oncology group. Arch Pathol Lab Med. 2015;139(10):1281–7. https://doi.org/10.5858/arpa.2014-0475-OA .CrossRefPubMedPubMedCentral

Esiashvili N, Prabhu R, Kahn S, Paulino AC. Current strategies and challenges in treatment of childhood rhabdomyosarcoma. J Radiat Oncol. 2013;2(2):159–68. https://doi.org/10.1007/s13566-012-0083-2 .CrossRef

McDowell HP, Foot ABM, Ellershaw C, Machin D, Giraud C, Bergeron C. Outcomes in paediatric metastatic rhabdomyosarcoma: results of the International Society of Paediatric Oncology (SIOP) study MMT-98. Eur J Cancer. 2010;46(9):1588–95. https://doi.org/10.1016/j.ejca.2010.02.051 .CrossRefPubMed

Oberlin O, Rey A, Lyden E, Bisogno G, Stevens MCG, Meyer WH, et al. Prognostic factors in metastatic Rhabdomyosarcomas: results of a pooled analysis from United States and European cooperative groups. J Clin Oncol. 2008;26(14):2384–9. https://doi.org/10.1200/JCO.2007.14.7207 .CrossRefPubMedPubMedCentral

Breneman JC, Lyden E, Pappo AS, Link MP, Anderson JR, Parham DM, et al. Prognostic factors and clinical outcomes in children and adolescents with metastatic rhabdomyosarcoma--a report from the intergroup Rhabdomyosarcoma study IV. J Clin Oncol. 2003;21(1):78–84. https://doi.org/10.1200/JCO.2003.06.129 .CrossRefPubMed

10.

Hoang NT, Acevedo LA, Mann MJ, Tolani B. A review of soft-tissue sarcomas: translation of biological advances into treatment measures. Cancer Manag Res. 2018;10:1089–114. https://doi.org/10.2147/CMAR.S159641 .CrossRefPubMedPubMedCentral

11.

Sorensen PHB, Lynch JC, Qualman SJ, Tirabosco R, Lim JF, Maurer HM, et al. PAX3-FKHR and PAX7-FKHR gene fusions are prognostic indicators in alveolar rhabdomyosarcoma: a report from the children's oncology group. J Clin Oncol. 2002;20(11):2672–9. https://doi.org/10.1200/JCO.2002.03.137 .CrossRefPubMed

12.

Wachtel M, Schäfer BW. PAX3-FOXO1: zooming in on an “undruggable” target. Semin Cancer Biol. 2018;50:115–23. https://doi.org/10.1016/j.semcancer.2017.11.006 .CrossRefPubMed

13.

Marshall AD, Grosveld GC. Alveolar rhabdomyosarcoma – the molecular drivers of PAX3/7-FOXO1-induced tumorigenesis. Skelet Muscle. 2012;2(1):25. https://doi.org/10.1186/2044-5040-2-25 .CrossRefPubMedPubMedCentral

14.

Stevens MCG. Treatment for childhood rhabdomyosarcoma: the cost of cure. Lancet Oncol. 2005;6(2):77–84. https://doi.org/10.1016/S1470-2045(05)01733-X .CrossRefPubMed

15.

Piranlioglu R, Lee E, Ouzounova M, Bollag RJ, Vinyard AH, Arbab AS, et al. Primary tumor-induced immunity eradicates disseminated tumor cells in syngeneic mouse model. Nat Commun. 2019;10(1):1430. https://doi.org/10.1038/s41467-019-09015-1 .CrossRefPubMedPubMedCentral

16.

Lambert AW, Pattabiraman DR, Weinberg RA. Emerging biological principles of metastasis. Cell. 2017;168(4):670–91. https://doi.org/10.1016/j.cell.2016.11.037 .CrossRefPubMedPubMedCentral

17.

Seyfried TN, Huysentruyt LC. On the origin of Cancer metastasis. Crit Rev Oncog. 2013;18(1 - 2):43–73. https://doi.org/10.1615/CritRevOncog.v18.i1-2.40 .CrossRefPubMedPubMedCentral

18.

Gandalovičová A, Rosel D, Fernandes M, Veselý P, Heneberg P, Čermák V, et al. Migrastatics—anti-metastatic and anti-invasion drugs: promises and challenges. Trends Cancer. 2017;3(6):391–406. https://doi.org/10.1016/j.trecan.2017.04.008 .CrossRefPubMedPubMedCentral

19.

Hall A. The cytoskeleton and cancer. Cancer Metastasis Rev. 2009;28(1-2):5–14. https://doi.org/10.1007/s10555-008-9166-3 .CrossRefPubMed

20.

Bonello TT, Stehn JR, Gunning PW. New approaches to targeting the actin cytoskeleton for chemotherapy. Future Med Chem. 2009;1(7):1311–31. https://doi.org/10.4155/fmc.09.99 .CrossRefPubMed

21.

Engel N, Ali I, Adamus A, Frank M, Dad A, Ali S, et al. Antitumor evaluation of two selected Pakistani plant extracts on human bone and breast cancer cell lines. BMC Complement Altern Med. 2016;16(1):244. https://doi.org/10.1186/s12906-016-1215-9 .CrossRefPubMedPubMedCentral

22.

Engel N, Kraft K, Müller P, Duske K, Kühn J, Oppermann C, et al. Actin cytoskeleton reconstitution in MCF-7 breast cancer cells initiated by a native flax root extract: Advancement in Medicinal Plant Research Vol. 3(3), pp. 92–105, July 2015 ISSN: 2354–2152 Full Length Research Paper. Adv Med Plant Res. 2015;3:92–105.

23.

Engel N, Lisec J, Piechulla B, Nebe B. Metabolic profiling reveals sphingosine-1-phosphate kinase 2 and lyase as key targets of (phyto-) estrogen action in the breast cancer cell line MCF-7 and not in MCF-12A. PLoS One. 2012;7(10):e47833. https://doi.org/10.1371/journal.pone.0047833 .CrossRefPubMedPubMedCentral

24.

Azrad M, Vollmer RT, Madden J, Dewhirst M, Polascik TJ, Snyder DC, et al. Flaxseed-derived Enterolactone is inversely associated with tumor cell proliferation in men with localized prostate Cancer. J Med Food. 2013;16(4):357–60. https://doi.org/10.1089/jmf.2012.0159 .CrossRefPubMedPubMedCentral

25.

Adolphe JL, Whiting SJ, Juurlink BHJ, Thorpe LU, Alcorn J. Health effects with consumption of the flax lignan secoisolariciresinol diglucoside. Br J Nutr. 2010;103(7):929–38. https://doi.org/10.1017/S0007114509992753 .CrossRefPubMed

26.

Thompson LU, Chen JM, Li T, Strasser-Weippl K, Goss PE. Dietary flaxseed alters tumor biological markers in postmenopausal breast cancer. Clin Cancer Res. 2005;11(10):3828–35. https://doi.org/10.1158/1078-0432.CCR-04-2326 .CrossRefPubMed

27.

Spagnuolo C, Russo GL, Orhan IE, Habtemariam S, Daglia M, Sureda A, et al. Genistein and Cancer: current status, challenges, and future Directions12. Adv Nutr. 2015;6(4):408–19. https://doi.org/10.3945/an.114.008052 .CrossRefPubMedPubMedCentral

28.

Borrás C, Gambini J, Gómez-Cabrera MC, Sastre J, Pallardó FV, Mann GE, et al. Genistein, a soy isoflavone, up-regulates expression of antioxidant genes: involvement of estrogen receptors, ERK1/2, and NFkappaB. FASEB J. 2006;20(12):2136–8. https://doi.org/10.1096/fj.05-5522fje .CrossRefPubMed

29.

Hassan G, Zahra Q, Engel N, Ali K, Bahreen G, Ahmad V, et al. Anticancer Activity of Hispidulin from Saussurea simpsoniana and Lupeol from Vincetoxicum arnottianum. TJNPR. 2020;4:31–5. https://doi.org/10.26538/tjnpr/v4i2.2 .CrossRef

30.

Adamus A, Peer K, Ali I, Lisec J, Falodun A, Frank M, et al. Berberis orthobotrys - a promising herbal anti-tumorigenic candidate for the treatment of pediatric alveolar rhabdomyosarcoma. J Ethnopharmacol. 2019;229:262–71. https://doi.org/10.1016/j.jep.2018.10.002 .CrossRefPubMed

31.

Adamus A, Engel N, Seitz G. SGPL1 321 mutation: one main trigger for invasiveness of pediatric alveolar rhabdomyosarcoma. Cancer Gene Ther. 2020;27(7-8):571–84. https://doi.org/10.1038/s41417-019-0132-8 .CrossRefPubMed

32.

Borowicz S, van Scoyk M, Avasarala S, Karuppusamy Rathinam MK, Tauler J, Bikkavilli RK, et al. The soft agar Colony formation assay. J Vis Exp. 2014;1(92):1. https://doi.org/10.3791/51998 .CrossRef

33.

Matschegewski C, Staehlke S, Birkholz H, Lange R, Beck U, Engel K, et al. Automatic Actin Filament Quantification of Osteoblasts and Their Morphometric Analysis on Microtextured Silicon-Titanium Arrays. Materials (Basel). 2012;5:1176–95. https://doi.org/10.3390/ma5071176 .CrossRef

34.

Adamus A, Müller P, Nissen B, Kasten A, Timm S, Bauwe H, et al. GCSH antisense regulation determines breast cancer cells’ viability. Sci Rep. 2018;8(1):15399. https://doi.org/10.1038/s41598-018-33677-4 .CrossRefPubMedPubMedCentral

35.

Lisec J, Schauer N, Kopka J, Willmitzer L, Fernie AR. Gas chromatography mass spectrometry-based metabolite profiling in plants. Nat Protoc. 2006;1(1):387–96. https://doi.org/10.1038/nprot.2006.59 .CrossRefPubMed

36.

Kopka J, Schauer N, Krueger S, Birkemeyer C, Usadel B, Bergmüller E, et al. GMD@CSB.DB: the Golm Metabolome database. Bioinformatics. 2005;21(8):1635–8. https://doi.org/10.1093/bioinformatics/bti236 .CrossRefPubMed

37.

Jaeger C, Hoffmann F, Schmitt CA, Lisec J. Automated annotation and evaluation of in-source mass spectra in GC/atmospheric pressure chemical ionization-MS-based metabolomics. Anal Chem. 2016;88(19):9386–90. https://doi.org/10.1021/acs.analchem.6b02743 .CrossRefPubMed

38.

Anderson RL, Balasas T, Callaghan J, Coombes RC, Evans J, Hall JA, et al. A framework for the development of effective anti-metastatic agents. Nat Rev Clin Oncol. 2019;16(3):185–204. https://doi.org/10.1038/s41571-018-0134-8 .CrossRefPubMed

39.

Shah SA, Shah NA, Ullah S, Alam MM, Badshah H, Ullah S, et al. Documenting the indigenous knowledge on medicinal flora from communities residing near Swat River (Suvastu) and in high mountainous areas in swat-Pakistan. J Ethnopharmacol. 2016;182:67–79. https://doi.org/10.1016/j.jep.2016.02.008 .CrossRefPubMed

40.

Nguyen TH, Barr FG. Therapeutic approaches targeting PAX3-FOXO1 and its regulatory and transcriptional pathways in Rhabdomyosarcoma. Molecules. 2018;23:11. https://doi.org/10.3390/molecules23112798 .CrossRef

41.

Kikuchi K, Tsuchiya K, Otabe O, Gotoh T, Tamura S, Katsumi Y, et al. Effects of PAX3-FKHR on malignant phenotypes in alveolar rhabdomyosarcoma. Biochem Biophys Res Commun. 2008;365(3):568–74. https://doi.org/10.1016/j.bbrc.2007.11.017 .CrossRefPubMed

42.

Bernasconi M, Remppis A, Fredericks WJ, Rauscher FJ, Schäfer BW. Induction of apoptosis in rhabdomyosarcoma cells through down-regulation of PAX proteins. Proc Natl Acad Sci U S A. 1996;93(23):13164–9. https://doi.org/10.1073/pnas.93.23.13164 .CrossRefPubMedPubMedCentral

43.

Locasale JW. Serine, glycine and the one-carbon cycle: cancer metabolism in full circle. Nat Rev Cancer. 2013;13(8):572–83. https://doi.org/10.1038/nrc3557 .CrossRefPubMedPubMedCentral

44.

Jain M, Nilsson R, Sharma S, Madhusudhan N, Kitami T, Souza AL, et al. Metabolite profiling identifies a key role for glycine in rapid cancer cell proliferation. Science. 2012;336(6084):1040–4. https://doi.org/10.1126/science.1218595 .CrossRefPubMedPubMedCentral

45.

Rivera J, Proia RL, Olivera A. The alliance of sphingosine-1-phosphate and its receptors in immunity. Nat Rev Immunol. 2008;8(10):753–63. https://doi.org/10.1038/nri2400 .CrossRefPubMedPubMedCentral

46.

Pyne NJ, Pyne S. Sphingosine 1-phosphate and cancer. Nat Rev Cancer. 2010;10(7):489–503. https://doi.org/10.1038/nrc2875 .CrossRefPubMed

47.

Selvam SP, Ogretmen B. Sphingosine kinase/sphingosine 1-phosphate signaling in cancer therapeutics and drug resistance. Handb Exp Pharmacol. 2013;1:3–27. https://doi.org/10.1007/978-3-7091-1511-4_1 .CrossRef

48.

Pyne NJ, El Buri A, Adams DR, Pyne S. Sphingosine 1-phosphate and cancer. Adv Biol Regul. 2018;68:97–106. https://doi.org/10.1016/j.jbior.2017.09.006 .CrossRefPubMed

49.

Takabe K, Paugh SW, Milstien S, Spiegel S. “Inside-out” signaling of Sphingosine-1-phosphate: therapeutic targets. Pharmacol Rev. 2008;60(2):181–95. https://doi.org/10.1124/pr.107.07113 .CrossRefPubMed

50.

Engel N, Adamus A, Frank M, Kraft K, Kühn J, Müller P, et al. First evidence of SGPL1 expression in the cell membrane silencing the extracellular S1P siren in mammary epithelial cells. PLoS One. 2018;13(5):e0196854. https://doi.org/10.1371/journal.pone.0196854 .CrossRefPubMedPubMedCentral

51.

Schneider G, Bryndza E, Abdel-Latif A, Ratajczak J, Maj M, Tarnowski M, et al. Bioactive lipids S1P and C1P are prometastatic factors in human rhabdomyosarcoma, and their tissue levels increase in response to radio/chemotherapy. Mol Cancer Res. 2013;11(7):793–807. https://doi.org/10.1158/1541-7786.MCR-12-0600 .CrossRefPubMedPubMedCentral

52.

Libura J, Drukala J, Majka M, Tomescu O, Navenot JM, Kucia M, et al. CXCR4-SDF-1 signaling is active in rhabdomyosarcoma cells and regulates locomotion, chemotaxis, and adhesion. Blood. 2002;100(7):2597–606. https://doi.org/10.1182/blood-2002-01-0031 .CrossRefPubMed

53.

Pressey JG, Haas MC, Pressey CS, Kelly VM, Parker JN, Gillespie GY, et al. CD133 Marks a Myogenically primitive subpopulation in Rhabdomyosarcoma cell lines that are relatively Chemoresistant but sensitive to mutant HSV. Pediatr Blood Cancer. 2012;60(1):45–52. https://doi.org/10.1002/pbc.24117 .CrossRefPubMedPubMedCentral

54.

Takashima S, Oka Y, Fujiki F, Morimoto S, Nakajima H, Nakae Y, et al. Syndecan-4 as a biomarker to predict clinical outcome for glioblastoma multiforme treated with WT1 peptide vaccine. Future Sci OA. 2016;2(4):FSO96. https://doi.org/10.4155/fsoa-2015-0008 .CrossRefPubMedPubMedCentral

55.

Dos SPB, Lima K, Kremer JL, da Conceicao BB, de Paula Mariani BM, da Silva JC, et al. Stathmin 1 is highly expressed and associated with survival outcome in malignant adrenocortical tumours. Investig New Drugs. 2020;38(3):899–908. https://doi.org/10.1007/s10637-019-00846-9 .CrossRef

56.

Ladner RD. The role of dUTPase and uracil-DNA repair in cancer chemotherapy. Curr Protein Pept Sci. 2001;2(4):361–70. https://doi.org/10.2174/1389203013380991 .CrossRefPubMed

57.

el-Hajj HH, Zhang H, Weiss B. Lethality of a dut (deoxyuridine triphosphatase) mutation in Escherichia coli. J Bacteriol. 1988;170:1069–75. https://doi.org/10.1128/jb.170.3.1069-1075.1988 .CrossRefPubMedPubMedCentral

Title: Vincetoxicum arnottianum modulates motility features and metastatic marker expression in pediatric rhabdomyosarcoma by stabilizing the actin cytoskeleton
Authors: Anna Adamus
Iftikhar Ali
Vasileios Vasileiadis
Luai Al-Hileh
Jan Lisec
Marcus Frank
Guido Seitz
Nadja Engel
Publication date: 01-12-2021
Publisher: BioMed Central
Keywords: Metastasis
Rhabdomyosarcoma
Published in: BMC Complementary Medicine and Therapies / Issue 1/2021
Electronic ISSN: 2662-7671
DOI: https://doi.org/10.1186/s12906-021-03299-x

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Vincetoxicum arnottianum modulates motility features and metastatic marker expression in pediatric rhabdomyosarcoma by stabilizing the actin cytoskeleton

Abstract

Background

Methods

Results

Conclusions

Graphical abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Graphical abstract

Please log in to get access to this content

Other articles of this Issue 1/2021

Antimicrobial activity of the volatile substances from essential oils

Phytochemical analysis, in-vitro anti-proliferative, anti-oxidant, anti-diabetic, and anti-obesity activities of Rumex rothschildianus Aarons. extracts

Trichosanthin cooperates with Granzyme B to restrain tumor formation in tongue squamous cell carcinoma

Communication and information needs about complementary and alternative medicine: a qualitative study of parents of children with cancer

Therapeutic effect of various ginsenosides on rheumatoid arthritis

Bushen Huoxue recipe attenuates early pregnancy loss via activating endometrial COX2-PGE2 angiogenic signaling in mice