Low-dose paclitaxel modulates tumour fibrosis in gastric cancer

Tsukada,Tomoya; Fushida,Sachio; Harada,Shinichi; Terai,Shiroh; Yagi,Yasumichi; Kinoshita,Jun; Oyama,Katsunobu; Tajima,Hidehiro; Ninomiya,Itasu; Fujimura,Takashi; Ohta,Tetsuo

doi:10.3892/ijo.2013.1801

Low-dose paclitaxel modulates tumour fibrosis in gastric cancer

Authors:
- Tomoya Tsukada
- Sachio Fushida
- Shinichi Harada
- Shiroh Terai
- Yasumichi Yagi
- Jun Kinoshita
- Katsunobu Oyama
- Hidehiro Tajima
- Itasu Ninomiya
- Takashi Fujimura
- Tetsuo Ohta
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Affiliations: Department of Gastroenterologic Surgery, Division of Cancer Medicine, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8641, Japan, Center for Biomedical Research and Education, School of Medicine, Kanazawa University, Kanazawa, Ishikawa 920-8641, Japan
Published online on: January 29, 2013 https://doi.org/10.3892/ijo.2013.1801
Pages: 1167-1174
Copyright: © Tsukada et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Various treatments have been used for peritoneal dissemination, which is the most common mode of metastasis in gastric cancer, but sufficiently good clinical outcomes have not yet been obtained because of the presence of rich fibrous components and acquired drug resistance. Epithelial-mesenchymal transition (EMT) is one of the major causes of tissue fibrosis and transforming growth factor-β (TGF-β) has a pivotal function in the progression of EMT. Smad proteins play an important role in the TGF-β signalling pathway. The TGF-β/Smad signalling pathway can be modulated by stabilising microtubules with paclitaxel (PTX). Here, we investigated whether paclitaxel can modulate TGF-β/Smad signalling in human peritoneal methothelial cells (HPMCs). To determine the cytostatic concentrations of antineoplastic agents in HPMCs, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed using PTX, 5-fluorouracil and cisplatin. The minimum concentration that caused significant inhibition of TGF-β1-induced morphological changes in human peritoneal methothelial cells on pre-treatment with PTX was 5 nM at 48 h (cell viability: 87.1±1.5%, P<0.01). The TGF-β signalling cascade and the status of various fibrous components were evaluated by immunofluorescence staining, real-time quantitative PCR and western blotting. TGF-β signalling induced morphological changes, α-SMA expression and collagen I synthesis in HPMCs and PTX treatment suppressed these EMT-like changes. Moreover, PTX treatment markedly suppressed Smad2 phosphorylation. These data suggest that at a low-dose, PTX can significantly suppress the TGF-β/Smad signalling pathway by inhibiting Smad2 phosphorylation in the human peritoneum and that this can reduce stromal fibrosis.

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1	Fushida S, Kinoshita J, Yagi Y, Funaki H, Kinami S, Ninomiya I, Fujimura T, Nishimura G, Kayahara M and Ohta T: Dual anti-cancer effects of weekly intraperitoneal docetaxel in treatment of advanced gastric cancer patients with peritoneal carcinomatosis: a feasibility and pharmacokinetic study. Oncol Rep. 19:1305–1310. 2008.
2	Shimada S, Tanaka E, Marutsuka T, Honmyo U, Tokunaga H, Yagi Y, Aoki N and Ogawa M: Extensive intraoperative peritoneal lavage and chemotherapy for gastric cancer patients with peritoneal free cancer cells. Gastric Cancer. 5:168–172. 2002. View Article : Google Scholar : PubMed/NCBI
3	Yonemura Y, Kawamura T, Nojima N, Bandou E, Keizou T, Fujita H, Michiwa Y, Fujimura T, Fushida S, Ajisaka H and Miwa K: Postoperative results of left upper abdominal evisceration for advanced gastric cancer. Hepatogastroenterology. 47:571–574. 2000.PubMed/NCBI
4	Koizumi W, Narahara H, Hara T, Takagane A, Akiya T, Takagi M, Miyashita K, Nishizaki T, Kobayashi O, Takiyama W, Toh Y, Nagaie T, Takagi S, Yamamura Y, Yanaoka K, Orita H and Takeuchi M: S-1 plus cisplatin versus S-1 alone for first-line treatment of advanced gastric cancer (SPIRITS trial): a phase III trial. Lancet Oncol. 9:215–221. 2008. View Article : Google Scholar : PubMed/NCBI
5	Shirao K, Boku N, Yamada Y, Yamaguchi K, Doi T, Takiuchi H, Nasu J, Nakamura K, Fukuda H and Ohtsu A: Randomized phase III study of 5-fluorouracil continuous infusion (5FUci) versus methotrexate and 5-FU sequential therapy (MF) in gastric cancer with peritoneal metastasis (JCOG0106). Proc ASCO. 27(Suppl 15): abs. 4545. 2009.
6	Tsukada T, Fushida S, Harada S, Yagi Y, Kinoshita J, Oyama K, Tajima H, Fujita H, Ninomiya I, Fujimura T and Ohta T: The role of human peritoneal mesothelial cells in the fibrosis and progression of gastric cancer. Int J Oncol. 41:476–482. 2012.PubMed/NCBI
7	Lv ZD, Na D, Ma XY, Zhao C, Zhao WJ and Xu HM: Human peritoneal mesothelial cell transformation into myofibroblasts in response to TGF-β1 in vitro. Int J Mol Med. 27:187–193. 2011.PubMed/NCBI
8	Hay ED: An overview of epithelio-mesenchymal transformation. Acta Anat. 154:8–20. 1995. View Article : Google Scholar : PubMed/NCBI
9	Kalluri R and Neilson EG: Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest. 112:1776–1784. 2003. View Article : Google Scholar : PubMed/NCBI
10	Thiery JP: Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2:442–454. 2002. View Article : Google Scholar : PubMed/NCBI
11	Nawshad A, Lagamba D, Polad A and Hay ED: Transforming growth factor-beta signaling during epithelial-mesenchymal transformation: implications for embryogenesis and tumor metastasis. Cells Tissues Organs. 179:11–23. 2005. View Article : Google Scholar
12	Miyazono K, Suzuki H and Imamura T: Regulation of TGF-beta signaling and its roles in progression of tumors. Cancer Sci. 94:230–234. 2003. View Article : Google Scholar : PubMed/NCBI
13	Derynck R and Zhang YE: Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature. 425:577–584. 2003. View Article : Google Scholar : PubMed/NCBI
14	Dong C, Li Z, Alvarez R Jr, Feng XH and Goldschmidt-Clermont PJ: Microtubule binding to Smads may regulate TGF beta activity. Mol Cell. 5:27–34. 2000. View Article : Google Scholar : PubMed/NCBI
15	Donaldson KL, Goolsby GL, Kiener PA and Wahl AF: Activation of p34cdc2 coincident with taxol-induced apoptosis. Cell Growth Differ. 5:1041–1050. 1994.PubMed/NCBI
16	Gelmon K: The taxoids: paclitaxel and docetaxel. Lancet. 344:1267–1272. 1994. View Article : Google Scholar : PubMed/NCBI
17	Ishigami H, Kitayama J, Kaisaki S, Hidemura A, Kato M, Otani K, Kamei T, Soma D, Miyato H, Yamashita H and Nagawa H: Phase II study of weekly intravenous and intraperitoneal paclitaxel combined with S-1 for advanced gastric cancer with peritoneal metastasis. Ann Oncol. 21:67–70. 2010. View Article : Google Scholar : PubMed/NCBI
18	Markman M, Bundy BN, Alberts DS, Fowler JM, Clark-Pearson DL, Carson LF, Wadler S and Sickel J: Phase III trial of standard-dose intravenous cisplatin plus paclitaxel versus moderately high-dose carboplatin followed by intravenous paclitaxel and intraperitoneal cisplatin in small-volume stage III ovarian carcinoma: an intergroup study of the Gynecologic Oncology Group, Southwestern Oncology Group and Eastern Cooperative Oncology Group. J Clin Oncol. 19:1001–1007. 2001.
19	Sakurai Y, Yoshida I, Tonomura S, Sakai W, Nakamura Y, Imazu H, Matsubara T and Ochiai M: Weekly administration of paclitaxel attenuated rectal stenosis caused by multiple peritoneal recurrence 8 years after the resection of gastric carcinoma. Gastric Cancer. 6:243–249. 2003.
20	Jordan MA, Toso RJ, Thrower D and Wilson L: Mechanism of mitotic block and inhibition of cell proliferation by taxol at low concentrations. Proc Natl Acad Sci USA. 90:9552–9556. 1993. View Article : Google Scholar : PubMed/NCBI
21	Axel DI, Kunert W, Göggelmann C, Oberhoff M, Herdeg C, Küttner A, Wild DH, Brehm BR, Riessen R, Köveker G and Karsch KR: Paclitaxel inhibits arterial smooth muscle cell proliferation and migration in vitro and in vivo using local drug delivery. Circulation. 96:636–645. 1997. View Article : Google Scholar : PubMed/NCBI
22	Colombo A, Drzewiecki J, Banning A, Grube E, Hauptmann K, Silber S, Dudek D, Fort S, Schiele F, Zmudka K and Guagliumi G: Randomized study to assess the effectiveness of slow- and moderate-release polymer-based paclitaxel-eluting stents for coronary artery lesions. Circulation. 108:788–794. 2003. View Article : Google Scholar
23	Stone GW, Ellis SG, Cox DA, Hermiller J, O’Shaughnessy C, Mann JT, Turco M, Caputo R, Bergin P, Greenberg J, Popma JJ and Russell ME; TAXUS-IV Investigators: A polymer-based, paclitaxel-eluting stent in patients with coronary artery disease. N Engl J Med. 350:221–231. 2004. View Article : Google Scholar : PubMed/NCBI
24	Liu X, Zhu S, Wang T, Hummers L, Wigley FM, Goldschmidt-Clermont PJ and Dong C: Paclitaxel modulates TGFbeta signaling in scleroderma skin grafts in immunodeficient mice. PLoS Med. 2:1334–1442. 2005.PubMed/NCBI
25	Choi HS, Savard CE, Choi JW, Kuver R and Lee SP: Paclitaxel interrupts TGF-beta1 signaling between gallbladder epithelial cells and myofibroblasts. J Surg Res. 141:183–191. 2007. View Article : Google Scholar : PubMed/NCBI
26	Yung S, Li FK and Chan TM: Peritoneal mesothelial cell culture and biology. Perit Dial Int. 26:162–173. 2006.PubMed/NCBI
27	Wakayama T, Sai Y, Ito A, Kato Y, Kurobo M, Murakami Y, Nakashima E, Tsuji A, Kitamura Y and Iseki S: Heterophilic binding of the adhesion molecules poliovirus receptor and immunoglobulin superfamily 4A in the interaction between mouse spermatogenic and Sertoli cells. Biol Reprod. 76:1081–1090. 2007. View Article : Google Scholar
28	Yagi Y, Fushida S, Harada S, Tsukada T, Kinoshita J, Oyama K, Fujita H, Ninomiya I, Fujimura T, Kayahara M, Kinuya S, Yashiro M, Hirakawa K and Ohta T: Biodistribution of humanized anti-VEGF monoclonal antibody/bevacizumab on peritoneal metastatic models with subcutaneous xenograft of gastric cancer in mice. Cancer Chemother Pharmacol. 66:745–753. 2010. View Article : Google Scholar
29	Yashiro M, Chung YS, Nishimura S, Inoue T and Sowa M: Fibrosis in the peritoneum induced by scirrhous gastric cancer cells may act as ‘soil’ for peritoneal dissemination. Cancer. 77:1668–1675. 1996.PubMed/NCBI
30	Nakazawa K, Yashiro M and Hirakawa K: Keratinocyte growth factor produced by gastric fibroblasts specifically stimulates proliferation of cancer cells from scirrhous gastric carcinoma. Cancer Res. 63:8848–8852. 2003.
31	Elenbaas B and Weinberg RA: Heterotypic signaling between epithelial tumor cells and fibroblasts in carcinoma formation. Exp Cell Res. 264:169–184. 2001. View Article : Google Scholar : PubMed/NCBI
32	Kinugasa S, Abe S, Tachibana M, Hishikawa Y, Yoshimura H, Monden N, Dhar DK, Nagasue N and Nagaoka S: Over expression of transforming growth factor-beta1 in scirrhous carcinoma of the stomach correlates with decreased survival. Oncology. 55:582–587. 1998. View Article : Google Scholar : PubMed/NCBI
33	Shinto O, Yashiro M, Kawajiri H, Shimizu K, Shimizu T, Miwa A and Hirakawa K: Inhibitory effect of a TGFbeta receptor type-I inhibitor, Ki26894, on invasiveness of scirrhous gastric cancer cells. Br J Cancer. 102:844–851. 2010. View Article : Google Scholar : PubMed/NCBI
34	Yashiro M, Chung YS and Sowa M: Role of orthotopic fibroblasts in the development of scirrhous gastric carcinoma. Jpn J Cancer Res. 85:883–886. 1994. View Article : Google Scholar : PubMed/NCBI
35	Inoue T, Chung YS, Yashiro M, Nishimura S, Hasuma T, Otani S and Sowa M: Transforming growth factor-beta and hepatocyte growth factor produced by gastric fibroblasts stimulate the invasiveness of scirrhous gastric cancer cells. Jpn J Cancer Res. 88:152–159. 1997. View Article : Google Scholar : PubMed/NCBI
36	Koyama T, Yashiro M, Inoue T, Nishimura S, Hirakawa YS and Chung K: TGF-beta1 secreted by gastric fibroblasts up-regulates CD44 H expression and stimulates the peritoneal metastatic ability of scirrhous gastric cancer cells. Int J Oncol. 16:355–362. 2000.PubMed/NCBI
37	Tamura T, Sasaki Y, Nishiwaki Y and Saijo N: Phase I study of paclitaxel by three-hour infusion: hypotension just after infusion is one of the major dose-limiting toxicities. Jpn J Cancer Res. 86:1203–1209. 1995. View Article : Google Scholar : PubMed/NCBI
38	Matsuoka H, Furusawa M, Tomoda H and Seo Y: Difference in cytotoxicity of paclitaxel against neoplastic and normal cells. Anticancer Res. 14:163–167. 1994.PubMed/NCBI
39	Gloushankova NA, Lyubimova AV, Tint IS, Feder HH, Vasiliev JM and Gelfand IM: Role of the microtubular system in morphological organization of normal and oncogene-transfected epithelial cells. Proc Natl Acad Sci USA. 91:8597–8601. 1994. View Article : Google Scholar : PubMed/NCBI
40	Schiff PB and Horwitz SB: Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci USA. 77:1561–1565. 1980. View Article : Google Scholar : PubMed/NCBI
41	Zhu S, Goldschmidt-Clermont PJ and Dong C: Transforming growth factor-β-induced inhibition of myogenesis is mediated through Smad pathway and is modulated by microtubule dynamic stability. Circ Res. 94:617–625. 2004.
42	Shi Y, Hata A, Lo RS, Massagué J and Pavletich NP: A structural basis for mutational inactivation of the tumour suppressor Smad4. Nature. 388:87–93. 1997. View Article : Google Scholar : PubMed/NCBI
43	Shi Y and Massague J: Mechanisms of TGF- signaling from cell membrane to the nucleus. Cell. 113:685–700. 2003. View Article : Google Scholar : PubMed/NCBI
44	Ten Dijke P, Goumans MJ, Itoh F and Itoh S: Regulation of cell proliferation by Smad proteins. J Cell Physiol. 191:1–16. 2002.PubMed/NCBI
45	Liu Q, Mao H, Nie J, Chen W, Yang Q, Dong X and Yu X: Transforming growth factor β1 induces epithelial-mesenchymal transition by activating the JNK-Smad3 pathway in rat peritoneal mesothelial cells. Perit Dial Int. 28:S88–S95. 2008.
46	Saika S, Yamanaka O, Ikeda K, Kim-Mitsuyama S, Flanders KC, Yoo J, Roberts AB, Nishikawa-Ishida I, Ohnishi Y, Muragaki Y and Ooshima A: Inhibition of p38MAP kinase suppresses fibrotic reaction of retinal pigment epithelial cells. Lab Invest. 85:838–850. 2005. View Article : Google Scholar : PubMed/NCBI
47	Wendling J, Marchand A, Mauviel A and Verrecchia F: 5-fluorouracil blocks transforming growth factor-beta-induced alpha 2 type I collagen gene (COL1A2) expression in human fibroblasts via c-Jun NH2-terminal kinase/activator protein-1 activation. Mol Pharmacol. 64:707–713. 2003. View Article : Google Scholar
48	Zhang D, Sun L, Xian W, Liu F, Ling G, Xiao L, Liu Y, Peng Y, Haruna Y and Kanwar YS: Low-dose paclitaxel ameliorates renal fibrosis in rat UUO model by inhibition of TGF-beta/Smad activity. Lab Invest. 90:436–447. 2010. View Article : Google Scholar : PubMed/NCBI