4-Methylumbelliferone inhibits enhanced hyaluronan synthesis and cell migration in pancreatic cancer cells in response to tumor-stromal interactions

Cheng,Xiao‑Bo; Sato,Norihiro; Kohi,Shiro; Koga,Atsuhiro; Hirata,Keiji

doi:10.3892/ol.2018.8147

4-Methylumbelliferone inhibits enhanced hyaluronan synthesis and cell migration in pancreatic cancer cells in response to tumor-stromal interactions

Authors:
- Xiao‑Bo Cheng
- Norihiro Sato
- Shiro Kohi
- Atsuhiro Koga
- Keiji Hirata
View Affiliations

Affiliations: Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, P.R. China, Department of Surgery 1, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Fukuoka 807‑8555, Japan
Published online on: March 1, 2018 https://doi.org/10.3892/ol.2018.8147
Pages: 6297-6301
Copyright: © Cheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Hyaluronic acid (HA) in tumor stroma promotes tumor invasion and progression. 4-Methylumbelliferone (4‑MU) is a potent HA synthesis inhibitor. In the present study, the effects of 4‑MU on enhanced HA synthesis and cell migration in pancreatic ductal adenocarcinoma (PDAC) cells, in response to co‑culture with stromal fibroblasts, was investigated. The HA concentration was determined using ELISA and a Transwell migration assay was used to analyze cell migratory capability. The mRNA expression levels of hyaluronan synthases (HAS1, HAS2 and HAS3) were determined using the quantitative polymerase chain reaction. Co‑culture between Panc‑1 cells and stromal fibroblasts markedly increased cell migration in association with increasing HA production, which was markedly associated with an increase in HAS3 mRNA expression. Treatment with 4‑MU markedly decreased the HA production and cell migration of Panc‑1 cells in the co‑culture system. The results of the present study suggested that interactions between PDAC cells and stromal fibroblasts increased HA production, resulting in a marked increase in migration of PDAC cells, and 4‑MU may be used as a chemotherapeutic agent to inhibit the enhanced migration of PDAC cells in response to tumor-stromal interactions.

View Figures

View References

1	Hidalgo M: Pancreatic cancer. N Engl J Med. 362:1605–1617. 2010. View Article : Google Scholar : PubMed/NCBI
2	Burris HA III, Moore MJ, Andersen J, Green MR, Rothenberg ML, Modiano MR, Cripps MC, Portenoy RK, Storniolo AM, Tarassoff P, et al: Improvements in survival and clinical benefit with gemcitabine as first-line therapy for patients with advanced pancreas cancer: A randomized trial. J Clin Oncol. 15:2403–2413. 1997. View Article : Google Scholar : PubMed/NCBI
3	Sultana A, Smith CT, Cunningham D, Starling N, Neoptolemos JP and Ghaneh P: Meta-analyses of chemotherapy for locally advanced and metastatic pancreatic cancer. J Clin Oncol. 25:2607–2615. 2007. View Article : Google Scholar : PubMed/NCBI
4	Moore MJ, Goldstein D, Hamm J, Figer A, Hecht JR, Gallinger S, Au HJ, Murawa P, Walde D, Wolff RA, et al: Erlotinib plus gemcitabine compared with gemcitabine alone in patients with advanced pancreatic cancer: A phase III trial of the national cancer institute of Canada clinical trials group. J Clin Oncol. 25:1960–1966. 2007. View Article : Google Scholar : PubMed/NCBI
5	Weigel PH and DeAngelis PL: Hyaluronan synthases: A decade-plus of novel glycosyltransferases. J Biol Chem. 282:36777–36781. 2007. View Article : Google Scholar : PubMed/NCBI
6	Tammi RH, Passi AG, Rilla K, Karousou E, Vigetti D, Makkonen K and Tammi MI: Transcriptional and post-translational regulation of hyaluronan synthesis. FEBS J. 278:1419–1428. 2011. View Article : Google Scholar : PubMed/NCBI
7	Entwistle J, Hall CL and Turley EA: HA receptors: Regulators of signalling to the cytoskeleton. J Cell Biochem. 61:569–577. 1996. View Article : Google Scholar : PubMed/NCBI
8	Adamia S, Maxwell CA and Pilarski LM: Hyaluronan and hyaluronan synthases: Potential therapeutic targets in cancer. Curr Drug Targets Cardiovasc Haematol Disord. 5:3–14. 2005. View Article : Google Scholar : PubMed/NCBI
9	Toole BP: Hyaluronan promotes the malignant phenotype. Glycobiology. 12:37R–42R. 2002. View Article : Google Scholar : PubMed/NCBI
10	Mahlbacher V, Sewing A, Elsässer HP and Kern HF: Hyaluronan is a secretory product of human pancreatic adenocarcinoma cells. Eur J Cell Biol. 58:28–34. 1992.PubMed/NCBI
11	Fries H, Elsässer HP, Mahlbacher V, Neumann K and Kern HF: Localisation of hyaluronate (HA) in primary tumors and nude mouse xenografts of human pancreatic carcinomas using a biotinylated HA-binding protein. Virchows Arch. 424:7–12. 1994. View Article : Google Scholar : PubMed/NCBI
12	McBride WH and Bard JB: Hyaluronidase-sensitive halos around adherent cells. Their role in blocking lymphocyte-mediated cytolysis. J Exp Med. 149:507–515. 1979. View Article : Google Scholar : PubMed/NCBI
13	Suzuki Y, Nishida Y, Naruse T, Gemba T and Ishiguro N: Pericellular matrix formation alters the efficiency of intracellular uptake of oligonucleotides in osteosarcoma cells. J Surg Res. 152:148–156. 2009. View Article : Google Scholar : PubMed/NCBI
14	Toole BP: Hyaluronan-CD44 interactions in cancer: Paradoxes and possibilities. Clin Cancer Res. 15:7462–7468. 2009. View Article : Google Scholar : PubMed/NCBI
15	Cheng XB, Sato N, Kohi S and Yamaguchi K: Prognostic impact of hyaluronan and its regulators in pancreatic ductal adenocarcinoma. PLoS One. 8:e807652013. View Article : Google Scholar : PubMed/NCBI
16	Nakamura T, Takagaki K, Shibata S, Tanaka K, Higuchi T and Endo M: Hyaluronic-acid-deficient extracellular matrix induced by addition of 4-methylumbelliferone to the medium of cultured human skin fibroblasts. Biochem Biophys Res Commun. 208:470–475. 1995. View Article : Google Scholar : PubMed/NCBI
17	Nakamura T, Funahashi M, Takagaki K, Munakata H, Tanaka K, Saito Y and Endo M: Effect of 4-methylumbelliferone on cell-free synthesis of hyaluronic acid. Biochem Mol Biol Int. 43:263–268. 1997.PubMed/NCBI
18	Kakizaki I, Takagaki K, Endo Y, Kudo D, Ikeya H, Miyoshi T, Baggenstoss BA, Tlapak-Simmons VL, Kumari K, Nakane A, et al: Inhibition of hyaluronan synthesis in Streptococcus equi FM100 by 4-methylumbelliferone. Eur J Biochem. 269:5066–5075. 2002. View Article : Google Scholar : PubMed/NCBI
19	Kudo D, Kon A, Yoshihara S, Kakizaki I, Sasaki M, Endo M and Takagaki K: Effect of a hyaluronan synthase suppressor, 4-methylumbelliferone, on B16F-10 melanoma cell adhesion and locomotion. Biochem Biophys Res Commun. 321:783–787. 2004. View Article : Google Scholar : PubMed/NCBI
20	Kakizaki I, Kojima K, Takagaki K, Endo M, Kannagi R, Ito M, Maruo Y, Sato H, Yasuda T, Mita S, et al: A novel mechanism for the inhibition of hyaluronan biosynthesis by 4-methylumbelliferone. J Biol Chem. 279:33281–33289. 2004. View Article : Google Scholar : PubMed/NCBI
21	Yoshihara S, Kon A, Kudo D, Nakazawa H, Kakizaki I, Sasaki M, Endo M and Takagaki K: A hyaluronan synthase suppressor, 4-methylumbelliferone, inhibits liver metastasis of melanoma cells. FEBS Lett. 579:2722–2726. 2005. View Article : Google Scholar : PubMed/NCBI
22	Morohashi H, Kon A, Nakai M, Yamaguchi M, Kakizaki I, Yoshihara S, Sasaki M and Takagaki K: Study of hyaluronan synthase inhibitor, 4-methylumbelliferone derivatives on human pancreatic cancer cell (KP1-NL). Biochem Biophys Res Commun. 345:1454–1459. 2006. View Article : Google Scholar : PubMed/NCBI
23	Nakazawa H, Yoshihara S, Kudo D, Morohashi H, Kakizaki I, Kon A, Takagaki K and Sasaki M: 4-methylumbelliferone, a hyaluronan synthase suppressor, enhances the anticancer activity of gemcitabine in human pancreatic cancer cells. Cancer Chemother Pharmacol. 57:165–170. 2006. View Article : Google Scholar : PubMed/NCBI
24	Vigetti D, Rizzi M, Viola M, Karousou E, Genasetti A, Clerici M, Bartolini B, Hascall VC, De Luca G and Passi A: The effects of 4-methylumbelliferone on hyaluronan synthesis, MMP2 activity, proliferation, and motility of human aortic smooth muscle cells. Glycobiology. 19:537–546. 2009. View Article : Google Scholar : PubMed/NCBI
25	Kultti A, Pasonen-Seppänen S, Jauhiainen M, Rilla KJ, Kärnä R, Pyöriä E, Tammi RH and Tammi MI: 4-Methylumbelliferone inhibits hyaluronan synthesis by depletion of cellular UDP-glucuronic acid and downregulation of hyaluronan synthase 2 and 3. Exp Cell Res. 315:1914–1923. 2009. View Article : Google Scholar : PubMed/NCBI
26	Lokeshwar VB, Lopez LE, Munoz D, Chi A, Shirodkar SP, Lokeshwar SD, Escudero DO, Dhir N and Altman N: Antitumor activity of hyaluronic acid synthesis inhibitor 4-methylumbelliferone in prostate cancer cells. Cancer Res. 70:2613–2623. 2010. View Article : Google Scholar : PubMed/NCBI
27	Twarock S, Freudenberger T, Poscher E, Dai G, Jannasch K, Dullin C, Alves F, Prenzel K, Knoefel WT, Stoecklein NH, et al: Inhibition of oesophageal squamous cell carcinoma progression by in vivo targeting of hyaluronan synthesis. Mol Cancer. 10:302011. View Article : Google Scholar : PubMed/NCBI
28	Urakawa H, Nishida Y, Wasa J, Arai E, Zhuo L, Kimata K, Kozawa E, Futamura N and Ishiguro N: Inhibition of hyaluronan synthesis in breast cancer cells by 4-methylumbelliferone suppresses tumorigenicity in vitro and metastatic lesions of bone in vivo. Int J Cancer. 130:454–466. 2012. View Article : Google Scholar : PubMed/NCBI
29	Nakamura R, Kuwabara H, Yoneda M, Yoshihara S, Ishikawa T, Miura T, Nozaka H, Nanashima N, Sato T and Nakamura T: Suppression of matrix metalloproteinase-9 by 4-methylumbelliferone. Cell Biol Int. 31:1022–1026. 2007. View Article : Google Scholar : PubMed/NCBI
30	Nakamura T, Ishikawa T, Nanashima N, Miura T, Nozaka H, Nakaoka R and Sato T: 4-Methylumbelliferone induces the expression of membrane type 1-matrix metalloproteinase in cultured human skin fibroblasts. Biochem Biophys Res Commun. 298:646–650. 2002. View Article : Google Scholar : PubMed/NCBI
31	Kuwabara H, Yoneda M, Hayasaki H, Nakamura T and Shibayama Y: A hyaluronan synthase suppressor, 4-methylumbelliferone, inhibits the tumor invasion associated with N-cadherin decreasement. Pathol Int. 61:262–263. 2011. View Article : Google Scholar : PubMed/NCBI
32	Holm Svensson AC, Bengtsson T, Grenegård M and Lindström EG: Hyaluronic acid influence on platelet-induced airway smooth muscle cell proliferation. Exp Cell Res. 318:632–640. 2012. View Article : Google Scholar : PubMed/NCBI
33	Twarock S, Tammi MI, Savani RC and Fischer JW: Hyaluronan stabilizes focal adhesions, filopodia, and the proliferative phenotype in esophageal squamous carcinoma cells. J Biol Chem. 285:23276–23284. 2010. View Article : Google Scholar : PubMed/NCBI
34	Cheng XB, Sato N, Kohi S, Koga A and Hirata K: Receptor for hyaluronic acid-mediated motility is associated with poor survival in pancreatic ductal adenocarcinoma. J Cancer. 6:1093–1098. 2015. View Article : Google Scholar : PubMed/NCBI
35	Knudson W, Biswas C and Toole BP: Interactions between human tumor cells and fibroblasts stimulate hyaluronate synthesis. Proc Natl Acad Sci USA. 81:6767–6771. 1984. View Article : Google Scholar : PubMed/NCBI
36	Sugahara KN, Murai T, Nishinakamura H, Kawashima H, Saya H and Miyasaka M: Hyaluronan oligosaccharides induce CD44 cleavage and promote cell migration in CD44-expressing tumor cells. J Biol Chem. 278:32259–32265. 2003. View Article : Google Scholar : PubMed/NCBI
37	Wu M, Cao M, He Y, Liu Y, Yang C, Du Y, Wang W and Gao F: A novel role of low molecular weight hyaluronan in breast cancer metastasis. FASEB J. 29:1290–1298. 2015. View Article : Google Scholar : PubMed/NCBI
38	Schmaus A, Klusmeier S, Rothley M, Dimmler A, Sipos B, Faller G, Thiele W, Allgayer H, Hohenberger P, Post S and Sleeman JP: Accumulation of small hyaluronan oligosaccharides in tumour interstitial fluid correlates with lymphatic invasion and lymph node metastasis. Br J Cancer. 111:559–567. 2014. View Article : Google Scholar : PubMed/NCBI
39	Fontaine L, Grand M, Molho D and Boschetti E: Spasmolytic activity of 4-methylumbelliferone on Oddi's sphincter. Studies on the mode of action of the drug. Therapie. 23:63–74. 1968.(In French). PubMed/NCBI
40	Fontaine L, Grand M, Molho D, Chabert MJ and Boschetti E: Choleretic, spasmolytic and general pharmacologic activities of 4-methylumbelliferone. Therapie. 23:51–62. 1968.(In French). PubMed/NCBI