Reduced expression of p16 and p27 proteins in nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) is a malignant tumor with a high incidence in east Asian countries. Inactivation of cyclin-dependent kinase (CDK) inhibitors (CKIs) and overexpression of G1 cyclin has been thought to be important for tumor development. To determine whether reduction of CKI (p16 and p27) expression was associated with NPC development, we performed immunohistochemical staining of NPC specimens from 20 patients. We found that p16 and p27 proteins were negative in 8 of 20 and 16 of 20 cases, respectively; that either p16 or p27 proteins were negative in 17 of 20; and that both p16 and p27 were negative in 7 of 20. Excepting the cases in which both CKIs were negative, negativity of p27 alone was statistically higher than that of p16 (9/20 versus 1/20, P = .022), suggesting that the reduction of p27 protein is an important event for the multi-step process of NPC development.

[Effect of combination chemotherapy with nedaplatin and 5-FU for head and neck squamous cell carcinoma].

Nedaplatin (254-S), which is a cisplatin (CDDP) analog, is an effective agent for head and neck squamous cell carcinoma (HNSCC). 254-S is expected to play an important role in neo-adjuvant chemotherapy (NAC) for HNSCC in place of CDDP. We have been using combination chemotherapy including CDDP, 5-FU, MTX and LV. The response rate and CR rate of this 4-drug combined chemotherapy are 87% and 33%. Thirty-six patients with HNSCC were treated with 5-FU, 800 mg/m2/day for 5 days and 254-S, 100 mg/m2 on day 4. Chemotherapy was discontinued in one patient because of allergic shock. Three patients showed a CR and 10 patients showed a PR. The response rate and CR rate of 254-S plus 5-FU chemotherapy were 37.1% and 8.6%. These were inferior to those with the 4-drug combined chemotherapy. Fourteen percent of patients showed grade 3 leukocytopenia, and 17% showed more than grade 3 thrombocytopenia. The effect of combination chemotherapy of 254-S and 5-FU was inferior to that of the previous chemotherapy including CDDP, 5-FU, MTX and LV. Further study or another combination therapy including 254-S will be essential for improving efficacy against HNSCC.

Cytostatic effect of inostamycin, an inhibitor of cytidine 5'-diphosphate 1,2-diacyl-sn-glycerol (CDP-DG): inositol transferase, on oral squamous cell carcinoma cell lines.

Inostamycin, which was recently isolated from Streptomyces sp. MH816-AF15 as an inhibitor of cytidine 5'-diphosphate 1,2-diacyl-sn-glycerol (CDP-DG): inositol transferase, caused a G1-phase accumulation in the cell cycle of small cell lung carcinomas. To investigate whether the cytostatic effect of inostamycin is restricted to lung carcinoma cell lines or applicable to other type of cells, we tested five oral squamous cell carcinoma (SCC) cell lines. Cell growth was suppressed in 62.5--125 ng/ml inostamycin in the culture medium in all oral cancer cell lines tested, with non-viable cells being <1%, indicating inostamycin is cytostatic on SCC cell lines. Decrease in cyclin D1 mRNA and protein expression due to the inostamycin treatment was accompanied by suppression of phosphorylated retinoblastoma susceptibility gene product (pRB-P) levels. Moreover, flow cytometric analysis showed that inostamycin induced an increase in G1/G0 cells (1.2--3.2 fold) over 24 h. These results suggest that inostamycin is a useful agent for tumour dormant cytostatic therapy for oral SCC.

Inostamycin, an inhibitor of cytidine 5'-diphosphate 1,2-diacyl-sn-glycerol (CDP-DG): inositol transferase, suppresses invasion ability by reducing productions of matrix metalloproteinase-2 and -9 and cell motility in HSC-4 tongue carcinoma cell line.

Inostamycin is an inhibitor of cytidine 5'-diphosphate 1,2-diacyl-sn-glycerol (CDP-DG): inositol transferase. It significantly reduced epidermal growth factor (EGF)-induced in vitro invasion of the tongue carcinoma cell line, HSC-4, through reconstituted basement membrane Matrigel. Since phosphatidylinositol (PI) 4,5-biphosphate is important for signal transduction through protein kinase C and actin reorganisation, we further examined the effect of inostamycin on production of two matrix metalloproteinases (MMPs), MMP-2 and -9, and on cell motility. Zymographic analysis showed that inostamycin suppressed pro-MMP-2 and pro-MMP-9 levels at a dose-dependent fashion, while MMP-2 activity was not significantly affected. By reverse transcription-polymerase chain reaction, it was found that inostamycin diminished steady state levels of MMP-2 and -9 but not membrane type 1-MMP mRNA expressions. Inostamycin partially blocked both EGF- and phorbol 12-myristate 13-acetate-stimulated pro-MMP-9 production. A cytoplasmic calcium chelator (BAPTA-AM) dramatically elevated pro- MMP-9 and slightly elevated pro-MMP-2 secretions. EGF-stimulated motility of HSC-4 cells was suppressed by inostamycin treatment along with reduction of actin cytoskeletal reorganisation, filopodia formation and cdc42 expression. These results suggested that inostamycin would be useful for an anti-invasive agent in tongue cancer.

[Basic studies on IL-2 gene therapy for head and neck carcinoma].

Basic studies on transduction of the interleukin-2 (IL-2) gene into tumor cells were carried out in order to develop a new immunotherapy for head and neck carcinomas. We transduced the IL-2 gene into KB cells using defective herpes simplex viral (HSV) amplicon vector as the gene transfer vehicle. A high level of IL-2 was produced by IL-2 gene transduced KB cells (KB/IL-2). Human peripheral blood mononuclear cells (PBMC) cultured in medium containing the culture supernatant of KB/IL-2 cells showed stronger cytotoxic activity against KB cells than the control. In in vivo studies, high levels of IL-2 and interferon-gamma (IFN-gamma) were detected in the serum of nude mice transplanted with KB/IL-2 cells. The spleen cells of KB/IL-2 cell-transplanted nude mice exhibited high cytotoxic activity. Three of 5 nude mice transplanted with KB/IL-2 cells were completely cured of their tumor, and all 3 mice survived for over 120 days. All 5 nude mice transplanted with KB/lacZ cells, and all 5 nude mice transplanted with KB cells died within 120 days as a result of tumor progression. In conclusion, transduction of IL-2 gene tumor cells was corroborated by the high level of IL-2 produced by KB/IL-2 cells. The mechanisms of tumor rejection on KB/IL-2 transplanted nude mice were thought to be that effector cells stimulated by IL-2 derived from transplanted KB/IL-2 cells killed tumor cells, and IFN-gamma produced by activated NK cells showed a synergistic effect on tumor cells-killing.

Expansion of the oral end of free revascularised jejunum with a jejunal patch flap rotated like a folding fan.

We have devised a new expansion process for the reconstruction of the cervical oesophagus with free revascularised jejunum in cases of hypopharyngeal cancer. The jejunal island flap is divided into three subislands and the major island of the caudal end is split partially at its oral end. The middle island is split, trimmed triangularly, and inserted into opened major island to expand the diameter of the oral end of the conduit for the cervical oesophagus and to make it funnel-shaped. The smallest island on the oral side is used as a circulation monitor for the transferred jejunum. Because the jejunal island patch is rotated around the root of mesentery like a folding fan and joined to the major island, there is neither shortening of the pedicle nor distortion. This method allows expansion of the oral end by up to 180% to facilitate end to end anastomosis in proximal pharyngeal defects.

[Thymidylate synthase expression in patients with hypopharyngeal carcinoma].

We analyzed the relationship between clinical response to neo-adjuvant chemotherapy including 5-fluorouracil (5-FU) in patients with hypopharyngeal carcinoma (HPC) and thymidylate synthase (TS) expression in their tumors. TS expression was evaluated with immunohistochemical staining techniques on biopsy specimens from HPC patients. TS immunostaining was divided into four levels (TS0-TS3) according to its level and pattern. The relationship between prognosis, tumor size, nodal status, differentiation of tumor cells and TS expression were also investigated. There was a statistically significant association between the level of TS expression and tumor size (p < 0.01). In terms of the effectiveness of chemotherapy, tumor differentiation, nodal status and prognosis, a statistical difference was not found in TS expression. These results suggest that the level of TS expression may show the degree of tumor proliferation, but may not necessarily be useful to obtain a response to chemotherapy including other drugs, e.g., cisplatin and other derivatives of platinum.

[Telomerase activity of tumors in the head and neck].

Telomerase is a maintaining enzyme of telomere repeats. Telemorase is expressed in immortal and cancer cells, but not in the normal tissues. We investigated telomerase activity of tumors in the head and neck. Telomerase activity was found in all of the cell lines derived from squamous cell carcinomas of the head and neck. Six of 9 (67%) primary fresh tissues from squamous cell carcinomas of the head and neck contained detectable telomerase activity. There were no apparent relationship between telomerase activity and clinical stage. Telomerase activity differed, depend on the responses to chemotherapy and radiotherapy: complete response, 0 of 3(0%); partial response; 0 of 1(0%); no change; 7 of 7 (100%). Therapy-resistant tumor seemed to have much telomerase activity and much malignant potential. In benign tumors, neither of 2 benign parotid tumors expressed telomerase activity, but one of 2(50%)papillomas had the telomerase activity. Telomerase is considered useful tumor marker for the diagnosis and prognosis.

[Postoperative swallowing function in tongue and oral floor cancer patients reconstructed with a recto-abdominal myocutaneous free flap after glossectomy--quantitative assessment of reconstructed tissue movement].

We examined the postoperative swallowing function of 12 tongue and oral floor cancer patients reconstructed with a recto-abdominal myocutaneous free flap after glossectomy. On the basis of the resection site, the present cases were classified into either anterior type or lateral type. Subjective evaluation of postoperative swallowing function was obtained from self-reports from patients. The movement of the reconstructed tissue was evaluated videofluorographically during swallowing, by tracking the movement of two small pellets temporarily attached to the anterior and central portions of the reconstructed tongue. The trajectory of the two pellets and the selected point of the hyoid bone were recorded together with the distance between the plate and the tongue dorsum, and between the posterior pharyngeal wall and the tongue base. Using a personal computer, a quantitative study of the video images was performed. The results were summarized as follows. 1) The movement pattern of the reconstructed tissue was generally saccadic rather than smooth. It was suggested that the swallowing pattern of the patients was different to that of normal controls. 2) Postoperative swallowing function was poorer in cases of the anterior type when compared to the lateral type. 3) In general, cases which showed relatively wide movement range appeared to achieve subjectively satisfactory swallowing function.

[Articulatory function in patients receiving glossectomy followed by reconstruction with a recto-abdominal myocutaneous free flap].

Postoperative articulatory functions of patients with tongue cancer have been improved by reconstructive surgery with a radial forearm or recto-abdominal myocutaneous free flap. We examined the postoperative articulatory functions of 10 patients who received reconstruction with a recto-abdominal myocutaneous free flap after glossectomy. The functions were investigated by standardized tests, i. e. a quentionnaires, the 100 Japanese monosyllable speech intelligibility test and a single-word intelligibility test. A confusion matrix was obtained from the results of the monosyllable test. On the basis of resection sites, the present cases were divided into two types: an anterior type and a lateral type. The results are summarized as follows. There was no significant difference in the results of the quentionnareis between the two types. The mean score of the 100 Japanese monosyllable speech intelligibility test in cases of the anterior type was 48% and in those of the lateral type it was 62%. The mean score of the single-word intellibibility test in cases of the anterior type was 75% and in those of the lateral type it was 83%. In cases of the anterior type, dental and alveolar sounds were often confused with fricatives, whereas in the lateral type, velars sounds were often confused with affricates or flaps. These results suggest that our classification based on resection site was useful for investigating postoperative articulatory functions after partial glossectomy.

[Case reports of mucosal melanoma of the head and neck].

Twelve patients with mucosal melanoma in the head and neck were treated at Yokohama city University from 1975 to 1994. The sex distribution was 5 male and 7 female, and the age ranged from 40 to 79 years-old. The highest number of patients were in their sixties. In 10 cases tumors arose in the nasal cavity, in one case in the maxillary sinus and in other case in the oral cavity. The treatment modality for this tumor consisted of various combination therapies including surgery, radiotherapy, chemotherapy and immunotherapy. Combination chemotherapy consisting of dimethyltriazeno imidazole carboxamide (DTIC, Dacarbazine), amino methyl pyrimidinyl methyl chlorethyl nitrorosourea hydrochloride (ACNU) and vincristine (VCR). Cisplatin (CDDP) was recently used for cases where other chemotherapy was not effective. Surgical treatment in the initial therapy was performed in 8 patients. In three of these 8 cases, the surgical margin was positive. One of them was dead after 9 months, another dead with complication after 1 year and the other survived free of any tumor. Immunotherapy using OK-432, interleukin-2, LAK therapy and low dose CPM was effective for some patients. The 5-year survival rate was 44%. Patients with surgical treatments in the initial therapy had longer survival than those without surgical treatments.

[Neuroendocrine carcinomas of the head and neck].

Neuroendocrine carcinomas (NECs) of the head and neck are rare. Between 1980 and 1995, only 163 cases of laryngeal and 30 cases of sinonasal NEC had been reported worldwide. We report two cases of sinonasal NEC. Case 1: A 49-year-old man complaining of left nasal obstruction was admitted to our hospital. A tumor with a tendency to bleed was seen in the left nasal cavity. CT scanning revealed an enhanced mass in the left nasal cavity and both maxillary and ethmoid sinuses. An open biopsy was performed to determine the diagnosis. The diagnosis of NEC was based on light microscopic and immunohistochemical findings. Chemotherapy and radiation were performed, but bone, liver and brain metastasis occurred 15 months after diagnosis and the patient died. Case 2: A 59-year-old man, complaining of right forehead swelling visited our department. CT scanning and MRI revealed that the right ethmoid sinus was filled by a tumor which had invaded the base of the skull. An open biopsy was performed. From the results of light microscopic and immunohistochemical investigations, NEC was highly suspected. Chemotherapy and radiation were performed, but the patient died 6 months after diagnosis. This study shows that there are aggressive types of nasal NEC, although these sinonasal carcinomas were reported in the literature to be not very lethal. The diagnosis, histopathological classification, treatment, and prognosis of sinonasal and laryngeal NECs were reviewed.

[A randomized crossover comparison of azasetron and granisetron in the prophylaxis of emesis induced by chemotherapy including cisplatin].

The clinical application of 5-HT3 receptor antagonists has enabled continuation of the course of chemotherapy including cisplatin, which induces strong nausea and vomiting, and to prevent the delay of curative treatment for cancer patients receiving neoadjuvant chemotherapy. However, with the development of basic research on the mechanisms of vomiting, each 5-HT3 receptor antagonist has appeared to have different pharmacological actions and, subsequently, the difference in the clinical efficacy of each drug has been reported in Europe and USA. In freshly advanced head and neck carcinoma cases, a randomised crossover study was performed to compare the efficacy and safety profile of a single intravenous dose for 7 days of azasetron (10 mg/day) or granisetron (3 mg/day) in the prophylaxis of nausea and vomiting induced by multi-drug chemotherapy including cisplatin (50 mg/m2 or 60 mg/m2). Anti-emetic effects were evaluated by the protective rates for nausea and vomiting for 7 days following the start of cisplatin administration. Both 5-HT3 receptor antagonists were highly effective in the prophylaxis of acute and delayed emesis induced by chemotherapy, whereas the efficacies of azasetron on day 3 and 4 were superior to those of granisetron. No adverse effect of either drug was observed in this study.

Sinusitis associated with disorders of visual acuity.

We report a case of sinusitis associated with disorders of visual acuity. The patient's vision was dramatically improved by sinus surgery despite the fact that the changes in the ethmoidal cells were slight and had no direct observable relation to the optic nerve canal. Possible mechanisms involved in nasal optic neuritis are discussed.

Secretion of a fibronectin-like substance from head and neck carcinomas.

The secretion of a fibronectin-like substance in culture supernatants of cell lines of head and neck squamous cell carcinomas was observed. Culture supernatants of some tumor cells had a high cell-attachment activity. Enzyme-linked immunosorbent assay with an antifibronectin antibody showed that the supernatant with cell attachment activity contained fibronectin. Furthermore, the cell attachment activity was clearly inhibited by the addition of a monoclonal antibody to fibronectin. These findings strongly suggest that the fibronectin produced by cancer cells is one of the potent mediators of the cell attachment.

Two cases of head and neck cancer with carotid artery reconstruction.

A carotid artery reconstruction was performed on two patients of head and neck cancer with infiltration into the carotid artery. To detect any possibility of cerebral ischemia at the time of ligation of the carotid artery, a temporary occlusion test of the internal carotid artery with a balloon catheter (balloon Matas test) was performed in both cases. An artificial vessel (Case 1) and a greater saphenous vein (Case 2) were used as an implanted vessel. No neurological sequelae were observed during or after the surgery in these cases. With a recent advance in various imaging diagnosis and an improvement in surgical techniques, the radical treatment including carotid artery reconstruction is expected to improve the prognosis for the cases demonstrating tumor infiltration into the carotid artery.

Modified combination chemotherapy of cisplatin and 5-fluorouracil in squamous cell carcinomas of the head and neck.

Randomized studies on the efficacy of two courses of different types of chemotherapy, including cisplatin and 5-fluorouracil (5-FU), were performed on 130 previously untreated cases with advanced squamous cell carcinomas of the head and neck. Cisplatin, followed by 120-hr continuous 5-FU infusion given in the conventional way, was administered to 60 patients (Group A), while cisplatin was administered 72 hr after the initiation of continuous 5-FU infusion in 70 other patients (Group B). The overall response rates (complete response plus partial response) were 58% in group A and 69% in group B, respectively. A superior complete response rate was obtained in cases receiving modified chemotherapy (10% in group A vs 20% in group B). There was no significant difference in the incidence of side effects between the two groups. These findings indicate that the modified cisplatin plus 5-FU combination chemotherapy tested here is more efficacious regimen than that of the conventional one to achieve high complete response rate and subsequently, to improve the survival of advanced carcinoma cases of the head and neck.

Induction chemotherapy in advanced head and neck cancer.

Induction chemotherapy, followed by definitive treatment, was performed in patients with advanced squamous-cell carcinoma of the head and neck. In this study, carried out between 1984 and 1991, testing the effectiveness of multimodality therapy in patients with previously untreated advanced (stage III and IV) squamous-cell carcinoma of the pharynx, patients received two different induction chemotherapy regimens: cisplatin, vincristine (Oncovin) plus peplomycin (COP), and cisplatin plus continuous 120-hr 5-fluorouracil (5-FU) infusion (CF) for two courses. Overall response rates (complete response plus partial response) to each of the two induction chemotherapy regimens were high: 76 and 82%, respectively. Superior complete response rate in the group receiving CF therapy was 16% versus 10% for COP therapy. Responders to induction chemotherapy had significantly better survival compared with non-responders. The toxicity of these two regimens was tolerable and manageable. It is indispensable to develop the more efficacious chemotherapy regimen with the potential to induce complete disappearance of tumors in patients with advanced head and neck carcinomas.

Tumours metastasizing to the head and neck--a report of seven cases.

Metastatic tumours involving the head and neck region are rare. Over the past 18 years, seven such cases were treated at our clinic. Of those, four were in one of the paranasal sinuses, three had arisen from a primary hepatocellular carcinoma and one from an osteogenic fibrosarcoma of the leg. In the remaining three cases, metastases to the larynx, the tonsil, and the parotid gland arose from a primary renal cell carcinoma, a thyroid carcinoma, and a breast carcinoma, respectively. In metastatic tumours, the primary site can often be identified by the histopathological features. Accordingly, when malignant head and neck tumours are suspected of being metastatic in character, it is important to search carefully for the primary site.

Extramedullary plasmacytoma of the larynx.

A case of IgD myeloma in a 54-year-old male with a long-standing history of extramedullary plasmacytoma involving the larynx is reported. The patient was treated with radiation therapy and laryngectomy. Twelve years later, he complained of nasal bleeding. On examination he was found to have large masses in the left nasal cavity and in the left supraclavicular region. Histological examination of both lesions showed plasmacytoma. Serum immunoglobulin studies revealed an IgD monoclonal spike of the lambda type. Bence-Jones protein was present. Using the immunoperoxidase staining technique, cytoplasmic monoclonal IgD was detected.