Cetuximab promotes anticancer drug toxicity in rhabdomyosarcomas with EGFR amplification in vitro

Yamamoto,Yuki; Fukuda,Kazumasa; Fuchimoto,Yasushi; Matsuzaki,Yumi; Saikawa,Yoshiro; Kitagawa,Yuko; Morikawa,Yasuhide; Kuroda,Tatsuo

doi:10.3892/or.2013.2588

Cetuximab promotes anticancer drug toxicity in rhabdomyosarcomas with EGFR amplification in vitro

Authors:
- Yuki Yamamoto
- Kazumasa Fukuda
- Yasushi Fuchimoto
- Yumi Matsuzaki
- Yoshiro Saikawa
- Yuko Kitagawa
- Yasuhide Morikawa
- Tatsuo Kuroda
View Affiliations

Affiliations: Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-858, Japan, Department of Surgery, Keio University School of Medicine, Tokyo 160-858, Japan, Division of Surgery, Department of Surgical Subspecialities, National Center for Child Health and Development, Tokyo 157-8535, Japan, Department of Physiology, Keio University School of Medicine, Tokyo 160-858, Japan
Published online on: July 4, 2013 https://doi.org/10.3892/or.2013.2588
Pages: 1081-1086

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

Overexpression of human epidermal growth factor receptor (EGFR) has been detected in various tumors and is associated with poor outcomes. Combination treatment regimens with EGFR-targeting and cytotoxic agents are a potential therapeutic option for rhabdomyosarcoma (RMS) with EGFR amplification. We investigated the effects of combination treatment with actinomycin D and the EGFR-targeting agent cetuximab in 4 RMS cell lines. All 4 RMS cell lines expressed wild-type K-ras, and 2 of the 4 overexpressed EGFR, as determined by flow cytometry, real-time PCR and direct sequencing. Combination of cetuximab and actinomycin D was highly effective, synergistically inhibiting cell growth with a combination index of less than 1. Moreover, combination treatment with these two reagents increased the rate of apoptosis in EGFR-positive cells. Cetuximab has antitumor activity in EGFR-amplified RMS cells when combined with antitumor reagents, indicating that cetuximab is a potential therapeutic reagent for RMS with EGFR amplification.

View Figures

View References

1	McDowell HP: Update on childhood rhabdomyosarcoma. Arch Dis Child. 88:354–357. 2003. View Article : Google Scholar
2	Pappo AS, Shapiro DN, Crist WM and Maurer HM: Biology and therapy of pediatric rhabdomyosarcoma. J Clin Oncol. 13:2123–2139. 1995.PubMed/NCBI
3	Wolden SL, Anderson JR, Crist WM, Breneman JC, Wharam MD Jr, Wiener ES, Qualman SJ and Donaldson SS: Indications for radiotherapy and chemotherapy after complete resection in rhabdomyosarcoma: a report from the Intergroup Rhabdomyosarcoma Studies I to III. J Clin Oncol. 17:3468–3475. 1999.PubMed/NCBI
4	Koscielniak E, Morgan M and Treuner J: Soft tissue sarcoma in children: prognosis and management. Paediatr Drugs. 4:21–28. 2002. View Article : Google Scholar : PubMed/NCBI
5	Ma WW and Adjei AA: Novel agents on the horizon for cancer therapy. CA Cancer J Clin. 59:111–137. 2009. View Article : Google Scholar : PubMed/NCBI
6	Capdevila J, Elez E, Macarulla T, Ramos FJ, Ruiz-Echarri M and Tabernero J: Anti-epidermal growth factor receptor monoclonal antibodies in cancer treatment. Cancer Treat Rev. 35:354–363. 2009. View Article : Google Scholar : PubMed/NCBI
7	Cunningham D, Humblet Y, Siena S, Khayat D, Bleiberg H, Santoro A, Bets D, Mueser M, Harstrick A, Verslype C, Chau I and Van Cutsem E: Cetuximab monotherapy and cetuximab plus irinotecan in irinotecan-refractory metastatic colorectal cancer. N Engl J Med. 351:337–345. 2004. View Article : Google Scholar : PubMed/NCBI
8	Herbst RS, Arquette M, Shin DM, Dicke K, Vokes EE, Azarnia N, Hong WK and Kies MS: Phase II multicenter study of the epidermal growth factor receptor antibody cetuximab and cisplatin for recurrent and refractory squamous cell carcinoma of the head and neck. J Clin Oncol. 23:5578–5587. 2005. View Article : Google Scholar : PubMed/NCBI
9	Vermorken JB: Squamous cell carcinoma of the head and neck. J BUON. 7:311–317. 2002.PubMed/NCBI
10	Peng D, Fan Z, Lu Y, DeBlasio T, Scher H and Mendelsohn J: Anti-epidermal growth factor receptor monoclonal antibody 225 up-regulates p27^KIP1 and induces G₁ arrest in prostatic cancer cell line DU145. Cancer Res. 56:3666–3669. 1996.PubMed/NCBI
11	Wu X, Fan Z, Masui H, Rosen N and Mendelsohn J: Apoptosis induced by an anti-epidermal growth factor receptor monoclonal antibody in a human colorectal carcinoma cell line and its delay by insulin. J Clin Invest. 95:1897–1905. 1995. View Article : Google Scholar : PubMed/NCBI
12	Perrotte P, Matsumoto T, Inoue K, Kuniyasu K, Eve BY, Hicklin DJ, Radinsky R and Dinney CPN: Anti-epidermal growth factor receptor antibody C225 inhibits angiogenesis in human transitional cell carcinoma growing orthotopically in nude mice. Clin Cancer Res. 5:257–265. 1999.PubMed/NCBI
13	Liu B, Fang M, Lu Y, Mendelsohn J and Fan Z: Fibroblast growth factor and insulin-like growth factor differentially modulate the apoptosis and G1 arrest induced by anti-epidermal growth factor receptor monoclonal antibody. Oncogene. 20:1913–1922. 2001. View Article : Google Scholar
14	Ciardiello F, Bianco R, Damiano V, De Lorenzo S, Pepe S, De Placido S, Fan Z, Mendelsohn J, Bianco AR and Tortora G: Antitumor activity of sequential treatment with topotecan and anti-epidermal growth factor receptor monoclonal antibody C225. Clin Cancer Res. 5:909–916. 1999.PubMed/NCBI
15	Kawaguchi Y, Kono K, Mimura K, Sugai H, Akaike H and Fujii H: Cetuximab induce antibody-dependent cellular cytotoxicity against EGFR-expressing esophageal squamous cell carcinoma. Int J Cancer. 120:781–787. 2007. View Article : Google Scholar
16	Kimura H, Sakai K, Arao T, Shimoyama T, Tamura T and Nishio K: Antibody-dependent cellular cytotoxicity of cetuximab against tumor cells with wild-type or mutant epidermal growth factor receptor. Cancer Sci. 98:1275–1280. 2007. View Article : Google Scholar
17	Kurai J, Chikumi H, Hashimoto K, Yamaguchi K, Yamasaki A, Sako T, Touge H, Makino H, Takata M, Miyata M, Nakamoto M, Burioka N and Shimizu E: Antibody-dependent cellular cytotoxicity mediated by cetuximab against lung cancer cell lines. Clin Cancer Res. 13:1552–1561. 2007. View Article : Google Scholar : PubMed/NCBI
18	Naramura M, Gillies SD, Mendelsohn J, Reisfeld RA and Mueller BM: Therapeutic potential of chimeric and murine anti-(epidermal growth factor receptor) antibodies in a metastasis model for human melanoma. Cancer Immunol Immunother. 37:343–349. 1993. View Article : Google Scholar : PubMed/NCBI
19	Chou TC and Talalay P: Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul. 22:27–55. 1984. View Article : Google Scholar : PubMed/NCBI
20	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
21	Sato O, Wada T, Kawai A, Yamaguchi U, Makimoto A, Kokai Y, Yamashita T, Chuman H, Beppu Y, Tani Y and Hasegawa T: Expression of epidermal growth factor receptor, ERBB2 and KIT in adult soft tissue sarcomas: a clinicopathologic study of 281 cases. Cancer. 103:1881–1890. 2005. View Article : Google Scholar : PubMed/NCBI
22	Ganti R, Skapek SX, Zhang J, Fuller CE, Wu J, Billups CA, Breitfeld PP, Dalton JD, Meyer WH and Khoury JD: Expression and genomic status of EGFR and ErbB-2 in alveolar and embryonal rhabdomyosarcoma. Mod Pathol. 19:1213–1220. 2006. View Article : Google Scholar : PubMed/NCBI
23	Margue CM, Bernasconi M, Barr FG and Schäfer BW: Transcriptional modulation of the anti-apoptotic protein BCL-XL by the paired box transcription factors PAX3 and PAX3/FKHR. Oncogene. 19:2921–2929. 2000. View Article : Google Scholar : PubMed/NCBI
24	Laé M, Ahn EH, Mercado GE, Chuai S, Edgar M, Pawel BR, Olshen A, Barr FG and Ladanyi M: Global gene expression profiling of PAX-FKHR fusion-positive alveolar and PAX-FKHR fusion-negative embryonal rhabdomyosarcomas. J Pathol. 212:143–151. 2007.PubMed/NCBI
25	Tomescu O, Xia SJ, Strezlecki D, Bennicelli JL, Ginsberg J, Pawel B and Barr FG: Inducible short-term and stable long-term cell culture systems reveal that the PAX3-FKHR fusion oncoprotein regulates CXCR4, PAX3, and PAX7 expression. Lab Invest. 84:1060–1070. 2004. View Article : Google Scholar : PubMed/NCBI
26	Taulli R, Scuoppo C, Bersani F, Accornero P, Forni PE, Miretti S, Grinza A, Allegra P, Schmitt-Ney M, Crepaldi T and Ponzetto C: Validation of Met as a therapeutic target in alveolar and embryonal rhabdomyosarcoma. Cancer Res. 66:4742–4749. 2006. View Article : Google Scholar : PubMed/NCBI
27	Mercado GE, Xia SJ, Zhang C, Ahn EH, Gustafson DM, Laé M, Ladanyi M and Barr FG: Identification of PAX3-FKHR-regulated genes differentially expressed between alveolar and embryonal rhabdomyosarcoma: focus on MYCN as a biologically relevant target. Genes Chromosomes Cancer. 47:510–520. 2008. View Article : Google Scholar
28	Williamson D, Lu YJ, Gordon T, Sciot R, Kelsey A, Fisher C, Poremba C, Anderson J, Pritchard-Jones K and Shipley J: Relationship between MYCN copy number and expression in rhabdomyosarcomas and correlation with adverse prognosis in the alveolar subtype. J Clin Oncol. 23:880–888. 2005. View Article : Google Scholar : PubMed/NCBI
29	Taniguchi E, Nishijo K, McCleish AT, Michalek JE, Grayson MH, Infante AJ, Abboud HE, Legallo RD, Qualman SJ, Rubin BP and Keller C: PDGFR-A is a therapeutic target in alveolar rhabdomyosarcoma. Oncogene. 27:6550–6560. 2008. View Article : Google Scholar : PubMed/NCBI
30	Davicioni E, Anderson MJ, Finckenstein FG, Lynch JC, Qualman SJ, Shimada H, Schofield DE, Buckley JD, Meyer WH, Sorensen PHB and Triche TJ: Molecular classification of rhabdomyosarcoma - genotypic and phenotypic determinants of diagnosis: a report from the Children’s Oncology Group. Am J Pathol. 174:550–564. 2009.PubMed/NCBI
31	Ebauer M, Wachtel M, Niggli FK and Schäfer BW: Comparative expression profiling identifies an in vivo target gene signature with TFAP2B as a mediator of the survival function of PAX3/FKHR. Oncogene. 26:7267–7281. 2007. View Article : Google Scholar : PubMed/NCBI
32	Oda Y, Kohashi K, Yamamoto H, Tamiya S, Kohno K, Kuwano M, Iwamoto Y, Tajiri T, Taguchi T and Tsuneyoshi M: Different expression profiles of Y-box-binding protein-1 and multidrug resistance-associated proteins between alveolar and embryonal rhabdomyosarcoma. Cancer Sci. 99:726–732. 2008. View Article : Google Scholar
33	Herrmann D, Seitz G, Warmann SW, Bonin M, Fuchs J and Armeanu-Ebinger S: Cetuximab promotes immunotoxicity against rhabdomyosarcoma in vitro. J Immunother. 33:279–286. 2010. View Article : Google Scholar : PubMed/NCBI
34	Lièvre A, Bachet J-B, Le Corre D, Boige V, Landi B, Emile JF, Côté JF, Tomasic G, Penna C, Ducreux M, Rougier P, Penault-Llorca F and Laurent-Puig P: KRAS mutation status is predictive of response to cetuximab therapy in colorectal cancer. Cancer Res. 66:3992–3995. 2006.PubMed/NCBI
35	Wellcome Trust Sanger Institute. Catalogue of Somatic Mutations in Cancer. http://www.sanger.ac.uk/genetics/CGP/cosmic/. Accessed June 4, 2012
36	Chen Y, Takita J, Hiwatari M, Igarashi T, Hanada R, Kikuchi A, Hongo T, Taki T, Ogasawara M, Shimada A and Hayashi Y: Mutations of the PTPN11 and RAS genes in rhabdomyosarcoma and pediatric hematological malignancies. Genes Chromosomes Cancer. 45:583–591. 2006. View Article : Google Scholar : PubMed/NCBI
37	Bancroft CC, Chen Z, Yeh J, Sunwoo JB, Yeh NT, Jackson S, Jackson C and Van Waes C: Effects of pharmacologic antagonists of epidermal growth factor receptor, PI3K and MEK signal kinases on NF-kappaB and AP-1 activation and IL-8 and VEGF expression in human head and neck squamous cell carcinoma lines. Int J Cancer. 99:538–548. 2002. View Article : Google Scholar : PubMed/NCBI
38	Raymond E, Faivre S and Armand JP: Epidermal growth factor receptor tyrosine kinase as a target for anticancer therapy. Drugs. 60(Suppl 1): 15–23. 2000. View Article : Google Scholar : PubMed/NCBI