title: The antitumor drug F14512 enhances cisplatin and ionizing radiation effects in head and neck squamous carcinoma cell lines.
abstract: BACKGROUND
  Head and Neck Squamous Cell Carcinoma (HNSCC) is the sixth most common cancer worldwide. The treatment of advanced stages HNSCC is based on surgical treatment combined with radiotherapy and chemotherapy or concomitant chemo-radiotherapy. However, the 5-year survival remains poor for advanced stages HNSCC and the development of new targeted therapies is eagerly awaited. F14512 combines an epipodophyllotoxin core-targeting topoisomerase II with a spermine moiety introduced as a cell delivery vector. This spermine moiety facilitates selective uptake by tumor cells via the Polyamine Transport System (PTS) and reinforces topoisomerase II poisoning. Here we report the evaluation of F14512 toward HNSCC.
  
  
  METHODS
  Four cell lines representative of head and neck cancer localizations were used: Fadu (pharynx), SQ20B (larynx), CAL33 and CAL27 (base of the tongue). PTS activity and specificity were evaluated by confocal microscopy and flow cytometry using the fluorescent probe F17073 which contains the same spermine moiety as F14512. Cytotoxicity, alone or in association with standard chemotherapeutic agents (cisplatin, 5FU), and radio-sensitizing effects were investigated using MTS and clonogenic assays, respectively. F14512 efficiency and PTS activity were also measured under hypoxic conditions (1% O2).
  
  
  RESULTS
  In all 4 tested HNSCC lines, an active PTS was evidenced providing a specific and rapid transfer of spermine-coupled compounds into cell nuclei. Interestingly, F14512 presents a 1.6-11-fold higher cytotoxic effect than the reference compound etoposide (lacking the spermine chain). It appears also more cytotoxic than 5FU and cisplatin in all cell lines. Competition experiments with spermine confirmed the essential role of the PTS in the cell uptake and cytotoxicity of F14512. Hypoxia had almost no impact on the drug cytotoxicity. The combination of F14512 with cisplatin, but not 5FU, was found to be synergistic and, for the first time, we demonstrated the significant radio-sensitizing potential of F14512.
  
  
  CONCLUSIONS
  The spermine moiety of F14512 confers a targeted effect and a much better efficacy than etoposide in HNSCC lines. The synergistic effect observed in association with cisplatin and radiotherapy augurs well for the potential development of F14512 in HNSCC.
authors.affiliation: Antitumoral Pharmacology Laboratory, Centre Oscar Lambret, Lille, France; ENT and Head and Neck Surgery Department, Huriez Hospital, University North of France, Lille, France; INSERM U837, Institute for Cancer Research of Lille (IRCL), Lille, France.
authors.forename: F
authors.initials: F
authors.lastname: Mouawad
authors.affiliation: Antitumoral Pharmacology Laboratory, Centre Oscar Lambret, Lille, France; INSERM U837, Institute for Cancer Research of Lille (IRCL), Lille, France.
authors.forename: A
authors.initials: A
authors.lastname: Gros
authors.affiliation: Antitumoral Pharmacology Laboratory, Centre Oscar Lambret, Lille, France; ENT and Head and Neck Surgery Department, Huriez Hospital, University North of France, Lille, France; INSERM U837, Institute for Cancer Research of Lille (IRCL), Lille, France.
authors.forename: B
authors.initials: B
authors.lastname: Rysman
authors.affiliation: Antitumoral Pharmacology Laboratory, Centre Oscar Lambret, Lille, France.
authors.forename: C
authors.initials: C
authors.lastname: Bal-Mahieu
authors.affiliation: Antitumoral Pharmacology Laboratory, Centre Oscar Lambret, Lille, France; INSERM U837, Institute for Cancer Research of Lille (IRCL), Lille, France.
authors.forename: C
authors.initials: C
authors.lastname: Bertheau
authors.affiliation: Antitumoral Pharmacology Laboratory, Centre Oscar Lambret, Lille, France; INSERM U837, Institute for Cancer Research of Lille (IRCL), Lille, France; Radiotherapy Department, Centre Oscar Lambret, Lille, France.
authors.forename: S
authors.initials: S
authors.lastname: Horn
authors.affiliation: Radiotherapy Department, Centre Oscar Lambret, Lille, France.
authors.forename: T
authors.initials: T
authors.lastname: Sarrazin
authors.affiliation: Radiotherapy Department, Centre Oscar Lambret, Lille, France.
authors.forename: E
authors.initials: E
authors.lastname: Lartigau
authors.affiliation: ENT and Head and Neck Surgery Department, Huriez Hospital, University North of France, Lille, France; INSERM U837, Institute for Cancer Research of Lille (IRCL), Lille, France.
authors.forename: D
authors.initials: D
authors.lastname: Chevalier
authors.affiliation: Research and Development Center Pierre Fabre, Toulouse, France.
authors.forename: C
authors.initials: C
authors.lastname: Bailly
authors.affiliation: Antitumoral Pharmacology Laboratory, Centre Oscar Lambret, Lille, France; INSERM U837, Institute for Cancer Research of Lille (IRCL), Lille, France.
authors.forename: A
authors.initials: A
authors.lastname: Lansiaux
authors.affiliation: Antitumoral Pharmacology Laboratory, Centre Oscar Lambret, Lille, France; INSERM U837, Institute for Cancer Research of Lille (IRCL), Lille, France. Electronic address: samuel.meignan@inserm.fr.
authors.forename: S
authors.initials: S
authors.lastname: Meignan
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000628:therapy; D002945:Cisplatin / Q000494:pharmacology; D005434:Flow Cytometry; D005472:Fluorouracil / Q000494:pharmacology; D006801:Humans; D007822:Laryngeal Neoplasms / Q000378:metabolism* / Q000628:therapy; D018613:Microscopy, Confocal; D010610:Pharyngeal Neoplasms / Q000378:metabolism* / Q000628:therapy; D011034:Podophyllotoxin / Q000031:analogs & derivatives* / Q000494:pharmacology; D014062:Tongue Neoplasms / Q000378:metabolism* / Q000628:therapy; D016896:Treatment Outcome
chemical_list: D000964:Antimetabolites, Antineoplastic; D000970:Antineoplastic Agents; C534191:F14512; D011034:Podophyllotoxin; D002945:Cisplatin; D005472:Fluorouracil
keywords: Etoposide; F14512; Head and neck cancer; Ionizing radiations; Polyamine transport system; Targeted therapy; Topoisomerase II inhibition