In vitro Monocyte IL-6 Secretion Levels Following Stimulation with Autologous Spheroids Derived from Tumour or Benign Mucosa Predict Long-term Survival in Head and Neck Squamous Cell Carcinoma Patients.

MCP-1/IL-6 in vitro monocyte secretion upon coculture with autologous fragment spheroids was studied in relation to patient 5- and 10-year overall survival rates in head and neck squamous cell carcinoma (HNSCC) patients (n = 65) diagnosed between 1998 and 2005, nine of whom had an human papilloma virus (HPV) tumour infection. The spheroids were harvested from malignant or benign tissue during primary surgery. Two weeks following surgery, freshly isolated autologous monocytes and benign or malignant spheroids were cocultured 24 h in vitro. The IL-6 secretion was expressed as a fraction of the lipopolysaccharide (LPS) response from the same batch of monocytes. HPV status was obtained by employing PCR analyses of primary diagnostic blocks. IL-6/MCP-1 response levels were not found to be dependent on HPV infection status. MCP-1 secretion did not predict prognosis, nor did in vitro IL-6 monocyte background or LPS-stimulated IL-6 secretion. At 5-year observation, dichotomized IL-6 levels following monocyte coculture, with both malignant and benign spheroids, showed a strong trend towards predicting survival, that is a low monocyte malignant coculture response showed a survival of 31 ± 17 versus 58 ± 17% with a high such response (P = 0.057). When studying monocyte IL-6 coculture responses evaluating benign and malignant spheroid results statistically together, a prediction of survival up to 10 years was found (hazard ratio = 0.48; confidence interval = 0.24-0.96; P < 0.05) with double low IL-6 responses. This survival prediction was also present after an adjustment for HPV tumour infection status. In conclusion, monocyte IL-6 in vitro secretion in cocultures with autologous spheroids/serum from HNSCCs predicted 5- and 10-year survivals, both with and without tumour HPV tumour adjustment.

Head and neck reconstruction using microsurgery: a 9-year retrospective study.

A total of 137 patients diagnosed with head and neck cancer underwent ablative surgery and primary free flap reconstruction during a period of 9 years, 2001-2009 (men 73.7 %, women 26.3 %). Surgery included a multidisciplinary approach involving plastic, head and neck, and maxillofacial surgeons. Squamous cell carcinoma (SCC) represented the vast majority of the diagnosed tumors (91.2 %); other tumors represented in the study were malignant melanoma, sarcoma, adenocarcinoma and blastoma. The free flaps (n = 143) used for reconstructive surgery included radial forearm flap (n = 128), fibular flap (n = 13) and rectus abdominis muscular flap (n = 2). Twenty patients (15 %) needed reoperation within 48 h due to clinical signs of hematoma (n = 8) and free flap ischemia (n = 12). Furthermore, we report a total of 12 free flap failures, giving an overall free flap success rate of 92 %. Five patients were treated due to infections at donor site (4 %). The overall survival rate (OS) in male patients diagnosed with oral SCC stage II-IV after 2 and 5 years was 82 and 78 %, respectively. Female patients in the same group displayed a 2- and 5-year OS of 78 and 67 %, respectively. Furthermore, analysis of patients treated for recurrence of primary SCC displayed a 2- and 5-year OS of 70 and 55 %, respectively. We conclude that our multidisciplinary approach and treatment algorithm for head and neck cancer including primary free flap reconstruction reconstitutes a safe and reliable tool.

Metoclopramide enhances the effect of ionizing radiation on xenografted squamous cell carcinoma of the head and neck.

The commonly used drug metoclopramide, a benzamide derivative, has been shown previously in our laboratory to enhance the effect of cisplatin on xenografted squamous cell carcinomas of the head and neck. In the present study, we show that metoclopramide also enhances the effect of ionizing radiation. Two human squamous cell carcinoma lines of the head and neck xenografted to nude mice have been used. Doses of radiation were chosen (5 and 8 Gy single doses) which caused only a slight retardation of tumor growth when administered alone. Tumor response to ionizing radiation was assessed with and without metoclopramide (2.0 mg kg-1), and administered at the time of radiation and 24 and 48 hr after treatment. The effects of these schedules on the tumors were compared using the reduction of the area under the growth curves and specific growth delay. The dose schedule with metoclopramide alone did not induce any significant reduction in the area under the growth curves. The addition of metoclopramide to the radiated groups caused a significant enhancement of the radiation-induced reduction of the area under the growth curves in both of the tumor lines studied.

Biochemical modulation of chemotherapy and radiotherapy in head and neck cancer.

In order to improve the treatment of advanced H&N cancer we must consider to adopt new strategies as: new/better cytostatic agents; new combinations of present cytostatic agents and; potentiation of radiotherapy and cytostatic agents by biochemical modulation, which we define as potentiation of therapy by non-chemotherapeutic agents. The list of agents that can potentiate chemotherapy or circumvene resistance is comprehensive. Most of the data are derived from in vitro studies. Much attention has been given to the fact that calcium channel blocker (CCB) agents can circumvene the multi-drug resistance (MDR) phenotype. Cisplatin, 5-fluorouracil, bleomycin and methotrexate, used in the treatment of head and neck cancer, are not part of the MDR phenotype. Still there are a few interesting reports indicating that CCB's could enhanced the antitumour actions of cisplatin, and that this interaction may be: (a) very specific; (b) unique to each species of CCB and (c) is independent of their binding affinity and classical function as inhibitors of the voltage sensitive calcium channels. Metoclopramide (MCA) is a structural analogue of procainamide used worldwide for preventing nausea and vomiting. It has structural resemblance to some of the known inhibitors of the DNA associated enzyme poly ADPRT such as benzamide. Benzamide is however rather toxic. MCA has been shown to enhance the effect of CDDP in vivo as well as in permeabilized cells in vitro, indicating that the DNA damaging effect of MCA is not dependent on cytoplasmatic enzymes or messenger systems. Radiobiologists have pointed out important biologic characteristics about tumour tissue such as hypoxia in relation to tumour radiosensitivity. Nicotinamide can effect tumour radiosensitivity in vivo. Comparing the response of mice in skin and tumour under different gas breathing regimens, a considerable therapeutic gain has been demonstrated for oxygen and carbon dioxide (95% O2 + 5% CO2) breathing mice. The effect of ionizing radiation (RT) on xenografted squamous cell carcinoma can also be enhanced by MCA. The optimal treatment interval is MCA given one hour before RT, which is in concordance with the hypothesis that MCA has to be present at the site of injury when RT is given in order to interact with repair mechanisms. We could not in conventional mice detect any MCA induced enhancement of either acute skin reaction or in LD50/30 after whole body irradiation, which indicates a potential therapeutic gain using MCA in a clinical setting. The above discussed interactions between biochemical modulators and chemo-/radio-therapeutic agents serve to illustrate the fruitful concept of biochemical potentiation of cytotoxicity.(ABSTRACT TRUNCATED AT 400 WORDS)

Increased response to cisplatin after long-term serial passage of a squamous cell carcinoma xenograft.

We have retrospectively investigated the response to cisplatin of a squamous cell carcinoma of the head and neck xenografted to nude mice during nine years of serial transplantation. Tumour growth rate decreased gradually. After nine years and over 100 passages, there was a sudden increase in cisplatin sensitivity. Histopathological examination showed that, of two histopathologically different subpopulations present in earlier passages, the predominant one was no longer detectable. The DNA-index did not change.

The amount of treatment versus quality of life in patients formerly treated for head and neck squamous cell carcinomas.

The aim of the present study was to investigate the association between the self-reported quality of life (QoL) versus the initial TNM stage and amount of primary and recurrent tumor therapy given in a population of formerly treated head and neck squamous cell carcinoma (HNSCC) patients. We determined QoL by the European Organization for Research and Treatment of Cancer-Quality of Life Questionnaire (EORTC-QLQ) C30/H&N35 by structured interview. One hundred and twenty-two patients less than 80 years old, who had been diagnosed with HNSCC in western Norway in the period from 1992 to1997, and who had survived until 2000, were identified. Of these patients, 106 were eligible to be included. Ninety-six of these patients agreed to be interviewed. For TNM stage as well as the type of therapy given (local surgery, neck dissection or radiation therapy), T stage predicted the general QoL scores. Both increased TNM stage and all given tumor therapy seemingly caused lower H&N symptom QoL scores. Of the various tumor treatments employed, neck radiation therapy and neck dissection were indicated to be the most closely associated with the H&N QoL scores. Having neck dissection performed seemingly caused impairment beyond what was explained by the initial TNM stage. In conclusion, tumor therapy to HNSCC should not be restricted due to general QoL considerations. Further study of how and when to perform neck treatment is suggested in order to avoid unnecessary reduced H&N QoL.

The benzamide derivative metoclopramide causes DNA damage and inhibition of DNA repair in human peripheral mononuclear leukocytes at clinically relevant doses.

The polysubstituted benzamide derivative metoclopramide (MCA) has previously been shown to enhance the effect of cisplatin and ionizing radiation treatment of xenografted human squamous cell carcinomas from the head and neck region. In the present work we show that MCA decreases the nucleoid sedimentation rate, indicating that MCA causes strand breaks in the DNA of human peripheral mononuclear leukocytes treated in vitro. This effect is seen with MCA in the dose range from 100 nM to 1 mM. MCA also stimulated the activity of the enzyme adenosine-diphospho-ribosyl transferase both in cells treated with MCA alone, and in combination with 15 Gy. This was taken as additional evidence that MCA causes DNA strand breaks. The DNA damage induced by MCA was poorly repaired when assessed by nucleoid sedimentation analysis, and this effect on repair was confirmed by showing that MCA also inhibits N-acetoxy-2-acetylaminofluorene-induced unscheduled DNA synthesis. The effect of MCA on DNA damage measured by nucleoid sedimentation has also been demonstrated in permeabilized cells. These data indicate that the DNA-damaging effect of MCA is not dependent on surface receptors or cytoplasmic processes.

Dose schedule evaluation of metoclopramide as a potentiator of cisplatin and carboplatin treatments of xenografted squamous cell carcinomas of the head and neck.

The potentiating effect of metoclopramide on the tumor growth inhibition of cisplatin has been studied on human squamous cell carcinomas xenografted to nude mice. In this system, the optimal time interval for intraperitoneal administration of metoclopramide was 8 h after intraperitoneal administration of cisplatin. The optimal single dose level of metoclopramide in this study was 2 mg/kg. Metoclopramide enhanced the cytotoxic effect of cisplatin at all cisplatin doses tested between 2.5 and 7.5 mg/kg body weight. Under experimental conditions that gave optimal sensitization of cisplatin-induced cytotoxicity, there was no potentiation of the cytotoxic effect with metoclopramide in combination with carboplatin. There is great similarity in the cytotoxic action of cisplatin and carboplatin, with the main difference being a much slower rate of formation of DNA crosslink formation following carboplatin exposure. Hence the data reported here support an important role for the kinetics of formation and reparability of DNA damage as part of the mechanism of metoclopramide sensitization of platinum-containing drugs.

Normal tissue reactions in mice after combined treatment with metoclopramide and ionizing radiation.

We have previously shown that metoclopramide potentiates the effect of ionizing radiation and cisplatin treatment of human squamous cell carcinomas from the head and neck region xenografted to nude mice. In the present tumor study, the dose scheduling of metoclopramide in combination with radiation was evaluated, and metoclopramide was shown to be most effective in potentiating the cytotoxic effect of radiation when administered one hour before radiation. The effect of radiation in combination with metoclopramide on normal tissue was also studied in two well-established models. Acute skin reactions to radiation exposure were studied in 129-type mice, and metoclopramide did not enhance the acute skin reaction in this in vivo model. Survival after whole body irradiation was studied in heterozygote Balb/c nu/+ mice as a measure of bone marrow toxicity. Metoclopramide was not found to affect the LD50/30 in this in vivo model. The absence of potentiation of radiation damage to normal tissue in these animal studies, makes metoclopramide an interesting possibility for future clinical evaluation.