Anticancer effects of NSC‑631570 (Ukrain) in head and neck cancer cells: In vitro analysis of growth, invasion, angiogenesis and gene expression

Herrmann,Ruth; Skaf,Joseph; Roller,Jeanette; Polednik,Christine; Holzgrabe,Ulrike; Schmidt,Marianne

doi:10.3892/or.2019.7416

Anticancer effects of NSC‑631570 (Ukrain) in head and neck cancer cells: In vitro analysis of growth, invasion, angiogenesis and gene expression

Authors:
- Ruth Herrmann
- Joseph Skaf
- Jeanette Roller
- Christine Polednik
- Ulrike Holzgrabe
- Marianne Schmidt
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Affiliations: Department of Otorhinolaryngology, University of Würzburg, D‑97080 Würzburg, Germany, Institute of Pharmacy and Food Chemistry, University of Würzburg, D‑97074 Würzburg, Germany
Published online on: November 25, 2019 https://doi.org/10.3892/or.2019.7416
Pages: 282-295

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Abstract

NSC‑631570 (Ukrain) is an aqueous extract of Chelidonium majus, a herbaceous perennial plant, one of two species in the genus Chelidonium, which has been demonstrated to selectively kill tumor cells without affecting non‑malignant cells. In the present study, the components of NSC‑631570 were examined by combined liquid chromatography/mass spectroscopy (LC‑MS) and the effects of NSC‑631570 on HNSCC cell lines, as well as primary cells, were analyzed with respect to growth, apoptosis, invasion, angiogenesis and gene expression. LC‑MS identified chelerythrine and allocryptopine as the major alkaloids of the extract. Moreover, NSC‑631570 suppressed the growth of all tested HNSCC cell lines, including a paclitaxel‑resistant and P‑glycoprotein (MDR1)‑overexpressing cell line. Mucosal keratinocytes were also affected by the extract, while fibroblasts proved to be much more resistant. In contrast to allocryptopine, chelerythrine had toxic effects on HNSCC cell lines at low doses. NSC‑631570 significantly induced apoptosis in the FaDu and HLaC78 cell lines. As analyzed by a spheroid‑based invasion assay, cell migration was significantly suppressed by NSC‑631570 in FaDu cells on gelatine, fibronectin, collagen, laminin and Matrigel®. Migration of the highly invasive cell line HLaC78 was also inhibited, albeit to a lesser extent (not significant on laminin). Microarray analysis revealed the downregulation of genes encoding key regulators, including EGFR, AKT2, JAK1, STAT3 and ß‑catenin (CTNNB1), all of which are involved in cell proliferation, migration, angiogenesis, apoptosis as well as the radiation‑ and chemo‑resistance of HNSCC. The strongest upregulation occurred for cytochrome P450 1A1 (CYP1A1) and 1B1 (CYP1B1), involved in the metabolism of xenobiotics. Upregulation of CYP1A1 was at least partially caused by chelerythrine and allocryptopine, as shown by RT‑qPCR in two HNSCC cell lines. In addition, NSC‑631570 showed a high anti‑angiogenic action on the tube formation ability of human umbilical vein endothelial cells (HUVECs). In conclusion, this study highlights NSC‑631570 as a promising therapeutic approach for HNSCC.

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1	Kamangar F, Dores GM and Anderson WF: Patterns of cancer incidence, mortality, and prevalence across five continents: Defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol. 24:2137–2150. 2006. View Article : Google Scholar : PubMed/NCBI
2	Goerner M, Seiwert TY and Sudhoff H: Molecular targeted therapies in head and neck cancer-an update of recent developments. Head Neck Oncol. 2:82010. View Article : Google Scholar : PubMed/NCBI
3	Hartwell JL: Plants Used Against Cancer: A Survey. Bioactive Plants Vol II. Quarterman Publications; Lawrence, MA: 1982
4	Danilos J, Zbroja-Sontag W, Baran E, Kurylcio L, Kondratowicz L and Jusiak L: Preliminary studies on the effect of Ukrain (Tris(2-([5bS-(5ba,6b,12ba)]-5b,6,7,12b,13,14-hexahydro-13-methyl[1,3] benzodioxolo[5,6-v]-1-3-dioxolo[4,5-i]phenanthridinium-6-ol]-ethaneaminyl)phosphinesulfide.6HCl) on the immunological response in patients with malignant tumours. Drugs Exp Clin Res. 18 (Suppl):S55–S62. 1992.
5	Hohenwarter O, Strutzenberger K, Katinger H, Liepins A and Nowicky JW: Selective inhibition of in vitro cell growth by the anti-tumour drug Ukrain. Drugs Exp Clin Res. 18 (Suppl):S1–S4. 1992.
6	Sotomayor EM, Rao K, Lopez DM and Liepins A: Enhancement of macrophage tumouricidal activity by the alkaloid derivative Ukrain. In vitro and in vivo studies. Drugs Exp Clin Res. 18 (Suppl):S5–S11. 1992.
7	Kadan P, Korsh OB and Hiesmayr W: Ukrain in the treatment of urethral recurrent carcinoma (case report). Drugs Exp Clin Res. 22:271–273. 1996.PubMed/NCBI
8	Lohninger A, Korsh OB and Melnyk A: Combined therapy with Ukrain and chemotherapy in ovarian cancer (case report). Drugs Exp Clin Res. 22:259–262. 1996.PubMed/NCBI
9	Schramm E, Nowicky JW and Godysh Y: Biophysiological effects of Ukrain therapy in a patient with breast cancer (case report). Drugs Exp Clin Res. 22:247–254. 1996.PubMed/NCBI
10	Habermehl D, Kammerer B, Handrick R, Eldh T, Gruber C, Cordes N, Daniel PT, Plasswilm L, Bamberg M, Belka C and Jendrossek V: Proapoptotic activity of Ukrain is based on Chelidonium majus L. alkaloids and mediated via a mitochondrial death pathway. BMC Cancer. 6:142006. View Article : Google Scholar : PubMed/NCBI
11	Lanvers-Kaminsky C, Nolting DM, Köster J, Schröder A, Sandkötter J and Boos J: In-vitro toxicity of Ukrain against human Ewing tumor cell lines. Anticancer Drugs. 17:1025–1030. 2006. View Article : Google Scholar : PubMed/NCBI
12	Panzer A, Hamel E, Joubert AM, Bianchi PC and Seegers JC: Ukrain(TM), a semisynthetic Chelidonium majus alkaloid derivative, acts by inhibition of tubulin polymerization in normal and malignant cell lines. Cancer Lett. 160:149–157. 2000. View Article : Google Scholar : PubMed/NCBI
13	Panzer A, Joubert AM, Bianchi PC and Seegers JC: The antimitotic effects of Ukrain, a Chelidonium majus alkaloid derivative, are reversible in vitro. Cancer Lett. 150:85–92. 2000. View Article : Google Scholar : PubMed/NCBI
14	Gagliano N, Moscheni C, Torri C, Donetti E, Magnani I, Costa F, Nowicky W and Gioia M: Ukrain modulates glial fibrillary acidic protein, but not connexin 43 expression, and induces apoptosis in human cultured glioblastoma cells. Anticancer Drugs. 18:669–676. 2007. View Article : Google Scholar : PubMed/NCBI
15	Herrmann R, Roller J, Polednik C and Schmidt M: Effect of chelidonine on growth, invasion, angiogenesis and gene expression in head and neck cancer cell lines. Oncol Lett. 16:3108–3116. 2018.PubMed/NCBI
16	Zenner HP, Lehner W and Herrmann IF: Establishment of carcinoma cell lines from larynx and submandibular gland. Arch Otorhinolaryngol. 225:269–277. 1979. View Article : Google Scholar : PubMed/NCBI
17	Schmidt M, Grünsfelder P and Hoppe F: Induction of matrix metalloproteinases in keratinocytes by cholesteatoma debris and granulation tissue extracts. Eur Arch Otorhinolaryngol. 257:425–429. 2000. View Article : Google Scholar : PubMed/NCBI
18	Vermes I, Haanen C, Steffens-Nakken H and Reutelingsperger C: A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods. 184:39–51. 1995. View Article : Google Scholar : PubMed/NCBI
19	Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U and Speed TP: Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics. 4:249–264. 2003. View Article : Google Scholar : PubMed/NCBI
20	Bolstad BM, Irizarry RA, Astrand M and Speed TP: A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics. 19:185–193. 2003. View Article : Google Scholar : PubMed/NCBI
21	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
22	El-Readi MZ, Eid S, Ashour ML, Tahrani A and Wink M: Modulation of multidrug resistance in cancer cells by chelidonine and Chelidonium majus alkaloids. Phytomedicine. 20:282–294. 2013. View Article : Google Scholar : PubMed/NCBI
23	Jesionek W, Fornal E, Majer-Dziedzic B, Móricz AM, Nowicky W and Choma IM: Investigation of the composition and antibacterial activity of Ukrain™ drug using liquid chromatography techniques. J Chromatogr A. 1429:340–347. 2016. View Article : Google Scholar : PubMed/NCBI
24	Kulp M and Bragina O: Capillary electrophoretic study of the synergistic biological effects of alkaloids from Chelidonium majus L. in normal and cancer cells. Anal Bioanal Chem. 405:3391–3397. 2013. View Article : Google Scholar : PubMed/NCBI
25	Malíková J, Zdarilová A, Hlobilková A and Ulrichová J: The effect of chelerythrine on cell growth, apoptosis, and cell cycle in human normal and cancer cells in comparison with sanguinarine. Cell Biol Toxicol. 22:439–453. 2006. View Article : Google Scholar : PubMed/NCBI
26	Mochly-Rosen D, Das K and Grimes KV: Protein kinase C, an elusive therapeutic target? Nat Rev Drug Discov. 11:937–957. 2012. View Article : Google Scholar : PubMed/NCBI
27	Jana J, Mondal S, Bhattacharjee P, Sengupta P, Roychowdhury T, Saha P, Kundu P and Chatterjee S: Chelerythrine downregulates expression of VEGFA, BCL2 and KRAS by arresting G-Quadruplex structures at their promoter regions. Sci Rep. 7:407062017. View Article : Google Scholar : PubMed/NCBI
28	Banerjee A, Sanyal S, Dutta S, Chakraborty P, Das PP, Jana K, Vasudevan M, Das C and Dasgupta D: The plant alkaloid chelerythrine binds to chromatin, alters H3K9Ac and modulates global gene expression. J Biomol Struct Dyn. 35:1491–1499. 2017. View Article : Google Scholar : PubMed/NCBI
29	Chmura SJ, Dolan ME, Cha A, Mauceri HJ, Kufe DW and Weichselbaum RR: In vitro and in vivo activity of protein kinase C inhibitor chelerythrine chloride induces tumor cell toxicity and growth delay in vivo. Clin Cancer Res. 6:737–742. 2000.PubMed/NCBI
30	Zhu Y, Pan Y, Zhang G, Wu Y, Zhong W, Chu C, Qian Y and Zhu G: Chelerythrine inhibits human hepatocellular carcinoma metastasis in vitro. Biol Pharm Bull. 41:36–46. 2018. View Article : Google Scholar : PubMed/NCBI
31	Koshelnick J, Moskvina E, Binder BR and Nowicky JW: Ukrain (NSC-631570) inhibits angiogenic differentiation of human endothelial cells in vitro. 17th International Cancer Congress Brazil: pp. 91–95. 1998
32	Rodriguez M and Potter DA: CYP1A1 regulates breast cancer proliferation and survival. Mol Cancer Res. 11:780–792. 2013. View Article : Google Scholar : PubMed/NCBI
33	Androutsopoulos VP, Tsatsakis AM and Spandidos DA: Cytochrome P450 CYP1A1: Wider roles in cancer progression and prevention. BMC Cancer. 9:1872009. View Article : Google Scholar : PubMed/NCBI
34	Orland A, Knapp K, König GM, Ulrich-Merzenich G and Knöß W: Combining metabolomic analysis and microarray gene expression analysis in the characterization of the medicinal plant Chelidonium majus L. Phytomedicine. 21:1587–1596. 2014. View Article : Google Scholar : PubMed/NCBI