Modulation of the expression of folate cycle enzymes and polyamine metabolism by berberine in cisplatin-sensitive and -resistant human ovarian cancer cells

Marverti,Gaetano; Ligabue,Alessio; Lombardi,Paolo; Ferrari,Stefania; Monti,Maria  Giuseppina; Frassineti,Chiara; Costi,Maria  Paola

doi:10.3892/ijo.2013.2045

Modulation of the expression of folate cycle enzymes and polyamine metabolism by berberine in cisplatin-sensitive and -resistant human ovarian cancer cells

Authors:
- Gaetano Marverti
- Alessio Ligabue
- Paolo Lombardi
- Stefania Ferrari
- Maria Giuseppina Monti
- Chiara Frassineti
- Maria Paola Costi
View Affiliations

Affiliations: Department of Biomedical Sciences, Metabolic and Neuroscience, Section of Pharmacology and Molecular Medicine, University of Modena and Reggio Emilia, I-41125 Modena, Italy, Naxospharma Srl, I-20026 Novate Milanese, Italy, Department of Life Sciences, University of Modena and Reggio Emilia, I-41125 Modena, Italy
Published online on: July 31, 2013 https://doi.org/10.3892/ijo.2013.2045
Pages: 1269-1280

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Berberine is a natural isoquinoline alkaloid with significant antitumor activity against many types of cancer cells, including ovarian tumors. This study investigated the molecular mechanisms by which berberine differently affects cell growth of cisplatin (cDDP)-sensitive and -resistant and polyamine analogue cross-resistant human ovarian cancer cells. The results show that berberine suppresses the growth of cDDP-resistant cells more than the sensitive counterparts, by interfering with the expression of folate cycle enzymes, dihydrofolate reductase (DHFR) and thymidylate synthase (TS). In addition, the impairment of the folate cycle also seems partly ascribable to a reduced accumulation of folate, a vitamin which plays an essential role in the biosynthesis of nucleic acids and amino acids. This effect was observed in both lines, but especially in the resistant cells, correlating again with the reduced tolerance to this isoquinoline alkaloid. The data also indicate that berberine inhibits cellular growth by affecting polyamine metabolism, in particular through the upregulation of the key catabolic enzyme, spermidine/spermine N1-acetyltransferase (SSAT). In this regard, berberine is shown to stimulate the SSAT induction by the spermine analogue N1, N12 bisethylspermine (BESpm), which alone was also able to downregulate DHFR mRNA more than TS mRNA. We report that the sensitivity of resistant cells to cisplatin or to BESpm is reverted to the levels of sensitive cells by the co-treatment with berberine. These data confirm the intimate inter-relationships between folate cycle and polyamine pathways and suggest that this isoquinoline plant alkaloid could be a useful adjuvant therapeutic agent in the treatment of ovarian carcinoma.

View Figures

View References

1.	Ozols RF, Bookman MA, Connolly DC, et al: Focus on 433 epithelial ovarian cancer. Cancer Cell. 5:19–24. 2004. View Article : Google Scholar
2.	Muggia F: Platinum compounds 30 years after the introduction of cisplatin: implications for the treatment of ovarian cancer. Gynecol Oncol. 112:275–281. 2009.PubMed/NCBI
3.	Jakubowicz-Gil J, Paduch R, Piersual T, Glowniak K, Gawron A and Kandefer M: The effect of quercetin on pro-apoptotic activity of cisplatin in HeLa cells. Biochem Pharmacol. 6:1343–1350. 2005. View Article : Google Scholar : PubMed/NCBI
4.	Maeda H, Hori S, Ohizumi H, Segawa T, Kakehi Y, Ogawa O and Kakizuka A: Effective treatment of advanced solid tumors by the combination of arsenic trioxide and L-buthionine-sulfoximine. Cell Death Differ. 11:737–746. 2004. View Article : Google Scholar : PubMed/NCBI
5.	Umanzor J and Aguiluz M: Concurrent cisplatin/gemcitabine chemotherapy along with radiotherapy in locally advanced cervical carcinoma: a phase II trial. Gynecol Oncol. 100:70–75. 2006. View Article : Google Scholar : PubMed/NCBI
6.	Scanlon KJ and Kashani-Sabet M: Elevated expression of thymidylate synthase cycle genes in cisplatin-resistant human ovarian carcinoma A2780 cells. Proc Natl Acad Sci USA. 85:650–653. 1988. View Article : Google Scholar : PubMed/NCBI
7.	Wysocki PJ: Targeted therapy of hepatocellular cancer. Expert Opin Investig Drugs. 19:265–274. 2010. View Article : Google Scholar : PubMed/NCBI
8.	Lin CC, Yang JS, Chen JT, et al: Berberine induces apoptosis in human HSC-3 oral cancer cells via simultaneous activation of the death receptor-mediated and mitochondrial pathway. Anticancer Res. 27:3371–3378. 2007.PubMed/NCBI
9.	Mantena SK, Sharma SD and Katiyar SK: Berberine, a natural product, induces G1-phase cell cycle arrest and caspase-3-dependent apoptosis in human prostate carcinoma cells. Mol Cancer Ther. 5:296–308. 2006. View Article : Google Scholar : PubMed/NCBI
10.	Yoon MJ, So HS, Cho HJ, et al: Berberine, a natural product, combined with cisplatin enhanced apoptosis through a mitochondria/caspase-mediated pathway in HeLa cells. Biol Pharm Bull. 31:789–795. 2008. View Article : Google Scholar : PubMed/NCBI
11.	Yi ZB, Yan Y, Liang YZ, et al: Evaluation of the antimicrobial mode of berberine by LC/ESI-MS combined with principal component analysis. J Pharm Biomed Anal. 44:301–304. 2007. View Article : Google Scholar : PubMed/NCBI
12.	Liu JC, Chan P, Chen YJ, et al: The antihypertensive effect of the berberine derivative 6-protoberberine in spontaneously hypertensive rats. Pharmacology. 59:283–289. 1999. View Article : Google Scholar : PubMed/NCBI
13.	Kuo CL, Chi CW and Liu TY: The anti-inflammatory potential of berberine in vitro and in vivo. Cancer Lett. 203:127–137. 2004. View Article : Google Scholar : PubMed/NCBI
14.	Un Y, Xun K, Wang Y, et al: A systematic review of the anti-cancer properties of berberine, a natural product from Chinese herbs. Anticancer Drugs. 20:757–769. 2009. View Article : Google Scholar : PubMed/NCBI
15.	Tang J, Feng Y, Tsao S, et al: Berberine and Coptidis rhizoma as novel antineoplastic agents: a review of traditional use and biomedical investigations. J Ethnopharmacol. 126:5172009.
16.	Kong B, Huang S, Wang W, May D, et al: Arsenic trioxide induces apoptosis in cisplatin-sensitive and -resistant ovarian cancer cell lines. Int J Gynecol Cancer. 15:872–877. 2005. View Article : Google Scholar : PubMed/NCBI
17.	Lin JP, Yang JS, Lee JH, Hsieh WT and Chung JG: Berberine induces cell cycle arrest and apoptosis in human gastric carcinoma SNU-5 cell line. World J Gastroenterol. 12:21–28. 2006.PubMed/NCBI
18.	Halestrap AP, Doran E, Gillespie JP and O’Toole A: Mitochondria and cell death. Biochem Soc Trans. 28:170–171. 2000.
19.	Duverger V, Sartorius U, Klein-Bauernschmitt P, Krammer PH and Schlehofer JR: Enhancement of cisplatin-induced apoptosis by infection with adeno-associated virus type 2. Int J Cancer. 97:706–712. 2002. View Article : Google Scholar : PubMed/NCBI
20.	Li JJ, Tang Q, Li Y, Hu BR, Ming ZY, Fu Q, Qian JQ and Xiang JZ: Role of oxidative stress in the apoptosis of hepatocellular carcinoma induced by combination of arsenic trioxide and ascorbic acid. Acta Pharmacol Sin. 27:1078–1084. 2006. View Article : Google Scholar : PubMed/NCBI
21.	Jantova S, Cipak L and Letasiova S: Berberine induces apoptosis through a mitochondrial/caspase pathway in human promonocytic U937 cells. Toxicol In Vitro. 21:25–31. 2007. View Article : Google Scholar : PubMed/NCBI
22.	Marverti G, Guaitoli G, Ligabue A, Frassineti C, Monti MG, Lombardi P and Costi MP: Distamycin A and derivatives as synergic drugs in cisplatin-sensitive and -resistant ovarian cancer cells. Amino Acids. 42:641–653. 2012. View Article : Google Scholar : PubMed/NCBI
23.	Arcamone F, Pencos PG, Orezzi PG, Nicolella V and Pirelli AM: Structure and synthesis of distamycin A. Nature. 203:1064–1065. 1964. View Article : Google Scholar : PubMed/NCBI
24.	Abu-Daya A and Fox KR: Interaction of minor groove binding ligands with long AT tracts. Nucleic Acids Res. 25:4962–4969. 1997. View Article : Google Scholar : PubMed/NCBI
25.	Carreras CW and Santi DV: The catalytic mechanism and structure of thymidylate synthase. Annu Rev Biochem. 64:721–762. 1995. View Article : Google Scholar : PubMed/NCBI
26.	Costi P and Ferrari S: Update on antifolate drugs targets. Curr Drug Targets. 2:135–166. 2001. View Article : Google Scholar : PubMed/NCBI
27.	Choi W, Gerner EW, Ramdas L, et al: Combination of 5-fluorouracil and N1,N11-Diethylnorspermine markedly activates spermidine/spermine N1-acetyltransferase expression, depletes polyamines and synergistically induces apoptosis in colon carcinoma cells. J Biol Chem. 280:3295–3304. 2005. View Article : Google Scholar
28.	Allen WL, McLean EG, Boyer J, et al: The role of spermidine/spermine N1-acetyltransferase in determining response to chemotherapeutic agents in colorectal cancer cells. Mol Cancer Ther. 6:128–137. 2007. View Article : Google Scholar : PubMed/NCBI
29.	Marverti G, Ligabue A, Guerrieri D, et al: Spermidine/spermine N1-acetyltranferase modulation by novel folate cycle inhibitors in cisplatin-sensitive and -resistant human ovarian cancer cell lines. Gynecol Oncol. 117:202–210. 2010. View Article : Google Scholar
30.	Wallace HM and Niiranen K: Polyamine analogues - an update. Amino Acids. 33:261–265. 2007. View Article : Google Scholar : PubMed/NCBI
31.	Wallace HM and Fraser AV: Inhibitors of polyamine metabolism: review article. Amino Acids. 26:353–365. 2004. View Article : Google Scholar : PubMed/NCBI
32.	Marverti G, Monti MG, Pegg AE, et al: Spermidine/spermine N1-acetyltransferase transient over-expression restores sensitivity of resistant human ovarian cancer cells to N1,N12-bis(ethyl)spermine and to cisplatin. Carcinogenesis. 26:1677–1686. 2005. View Article : Google Scholar
33.	Andrews PA, Murphy MP and Howell SB: Differential potentiation of alkylating and platinating agent cytotoxicity in human ovarian carcinoma cells by glutathione depletion. Cancer Res. 45:6250–6253. 1985.PubMed/NCBI
34.	Rossi T, Coppi A, Bruni E, Ruberto A, Santachiara S and Baggio G: Effects of anti-malarial drugs on MCF-7 and Vero cell replication. Anticancer Res. 27:2555–2560. 2007.PubMed/NCBI
35.	Lowry OH, Rosebrough NJ, Farr AL and Randall RJ: Protein measurement with the Folic phenol reagent. J Biol Chem. 193:265–275. 1951.PubMed/NCBI
36.	Kueng W, Siber E and Eppenberger U: Quantification of cells cultured on 96-well plates. Anal Biochem. 182:16–19. 1989. View Article : Google Scholar : PubMed/NCBI
37.	van Triest B, Pinedo HM, van Hensbergen Y, et al: Thymidylate synthase level as the main predictive parameter for sensitivity to 5-fluorouracil, but not for folate-based thymidilate synthase inhibitors, in 13 nonselected colon cancer cell lines. Clin Cancer Res. 5:643–654. 1999.
38.	Rothenberg SP, Perwaiz Iqbal M and Da Costa M: A simplified radioenzymatic assay for dihydrofolate reductase using [³H]dihydrofolate. Anal Biochem. 103:152–156. 1980.PubMed/NCBI
39.	Bradford MM: A rapid and sensitive method for quantification of microgram quantities of protein utilising the principle of protein-dye binding. Anal Biochem. 72:248–254. 1976. View Article : Google Scholar : PubMed/NCBI
40.	Arocho A, Chen B, Ladanyi M and Pan Q: Validation of the 2^−ΔΔCt calculation as an alternate method of data analysis for quantitative PCR of BCR-ABL P210 transcripts. Diagn Mol Pathol. 15:56–61. 2006.
41.	Kansara V, Paturi D, Luo S, Gaudana R and Mitra AK: Folic acid transport via high affinity carrier-mediated system in human retinoblastoma cells. Int J Pharm. 355:210–219. 2008. View Article : Google Scholar : PubMed/NCBI
42.	Casero RA Jr, Gabrielson EW and Pegg AE: Immunohistochemical staining of spermidine/spermine N1-acetyltransferase super-induced in response to treatment with antitumour polyamine analogues. Cancer Res. 54:3955–3958. 1994.
43.	Chou TC and Talalay P: Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Advances in Enzyme Regulation. Weber G: Pergamon Press; New York, NY: pp. 27–55. 1984, View Article : Google Scholar : PubMed/NCBI
44.	Marverti G, Ligabue A, Paglietti G, et al: Collateral sensitivity to novel thymidylate synthase inhibitors correlates with folate cycle enzymes impairment in cisplatin-resistant human ovarian cancer cells. Eur J Pharmacol. 615:17–26. 2009. View Article : Google Scholar
45.	Chu E and Allegra CJ: The role of thymidylate synthase as an RNA binding protein. Bioessays. 18:191–198. 1996. View Article : Google Scholar : PubMed/NCBI
46.	Marverti G, Piccinini G, Ghiaroni S, Barbieri D, Quaglino D and Moruzzi MS: N1,N12-bis(ethyl)spermine effect on growth of cis-diamminedichloroplatinum(II)-sensitive and -resistant human ovarian carcinoma cell lines. Int J Cancer. 78:33–40. 1998.PubMed/NCBI
47.	Orfila L, Rodrıguez M, Colman T, Hasegawa M, Merentes E and Arvelo F: Structural modification of berberine alkaloids in relation to cytotoxic activity in vitro. J Ethnopharmacol. 71:449–456. 2000. View Article : Google Scholar : PubMed/NCBI
48.	Gong GQ, Zong ZX and Song YM: Spectrofluorometric determination of DNA and RNA with berberine. Spectrochimica Acta A Mol Biomol Spectrosc. 55:1903–1907. 1999. View Article : Google Scholar : PubMed/NCBI
49.	Debnath D, Suresh Kumar G, Nandi R and Maiti M: Interaction of berberine chloride with deoxyribonucleic acids: evidence for base and sequence specificity. Indian J Biochem Biophys. 26:201–208. 1989.PubMed/NCBI
50.	Bhadra K, Maiti M and Suresh Kumar G: Berberine-DNA complexation: new insights into the cooperative binding and energetic aspects. Biochim Biophys Acta. 1780:1054–1061. 2008. View Article : Google Scholar : PubMed/NCBI
51.	Tan W, Li Y, Chen M and Wang Y: Berberine hydrochloride: anticancer activity and nanoparticulate delivery system. Int J Nanomedicine. 6:1773–1777. 2011. View Article : Google Scholar : PubMed/NCBI
52.	Maiti M and Kumar GS: Polymorphic nucleic acid binding of bioactive isoquinoline alkaloids and their role in cancer. J Nucleic Acids. 2010:1–23. 2010. View Article : Google Scholar : PubMed/NCBI
53.	Bhadra K and Kumar GS: Therapeutic potential of nucleic acid-binding isoquinoline alkaloids: binding aspects and implications for drug design. Med Res Rev. 31:821–862. 2011. View Article : Google Scholar : PubMed/NCBI
54.	Ferraroni M, Bazzicalupi C, Bilia AR and Gratteri P: X-ray diffraction analyses of the natural isoquinoline alkaloids Berberine and Sanguinarine complexed with double helix DNA d(CGTACG). Chem Commun. 47:4917–4919. 2011. View Article : Google Scholar : PubMed/NCBI
55.	Bhowmik D, Hossain M, Buzzetti F, D’Auria R, Lombardi P and Kumar GS: Biophysical studies on the effect of the 13 position substitution of the anticancer alkaloid berberine on its DNA binding. J Phys Chem B. 116:2314–2324. 2012. View Article : Google Scholar : PubMed/NCBI
56.	Cho J and Rando RR: Specific binding of Hoechst 33258 to site 1 thymidylate synthase mRNA. Nucleic Acids Res. 28:2158–2163. 2000. View Article : Google Scholar : PubMed/NCBI
57.	Mazzini S, Bellucci MC and Mondelli R: Mode of binding of the cytotoxic alkaloid berberine with the double helix oligo-nucleotide d(AAGAATTCTT)₂. Bioorg Med Chem. 11:505–514. 2003. View Article : Google Scholar : PubMed/NCBI
58.	Hiraku Y, Oikawa S and Kawanishi S: Distamycin A, a minor groove binder, changes enediyne-induced DNA cleavage sites and enhances apoptosis. Nucleic Acids Res (Suppl). 2:95–96. 2002. View Article : Google Scholar : PubMed/NCBI
59.	Basu S, Feuerstein BG, Dennis FD, Lubich WP, Bergeron RJ, Samejima K and Marton LJ: Correlation between the effects of polyamine analogues on DNA conformation and cell growth. Cancer Res. 49:5591–5597. 1989.PubMed/NCBI
60.	Agostinelli E, Arancia G, Dalla Vedova L, Belli F, Marra M, Salvi M and Toninello A: The biological functions of polyamine oxidation products by amine oxidases: perspectives of clinical applications. Amino Acids. 27:347–358. 2004. View Article : Google Scholar : PubMed/NCBI
61.	Sun D, Wollin A and Stephen AM: Moderate folate deficiency influences polyamine synthesis in rats. J Nutr. 132:2632–2637. 2002.PubMed/NCBI