Oncogenes: their presence and significance in squamous cell cancer of the head and neck

DNA extracted from squamous cell carcinomas of the larynx and tongue has been shown to contain cellular transforming genes characterized by their ability to transform mouse fibroblasts into malignant foci of cells which, when subsequently cloned and grown to volume, have been found to contain human DNA sequences. This DNA has been serially passaged through subsequent populations of NIH/3T3 mouse fibroblasts. Higher malignant transformation efficiencies have been observed and reported with serial passage. Of greater significance is the repeated identification of oncogenes of identical characteristics on electrophoretic radioisotope analysis.     

Verrucous carcinoma of the larynx. Possible human papillomavirus etiology

Verrucous carcinoma of the larynx is a distinct and uncommon variant of well-differentiated squamous cell carcinoma. By DNA hybridization techniques, we clearly demonstrated human papillomavirus (HPV-16-related) sequences in five patients with this neoplasm. In addition, HPV-16-related sequences were found in adjacent normal tissues. The DNAs from squamous cell carcinomas of the larynx were negative when hybridized to HPV-6, -11, or -16. Postirradiation anaplastic transformation of verrucous carcinoma has been described. We believe that radiotherapy should not be given unless the potential consequences are fully explained because of its potential to activate or alter HPV-16-related sequences.     

Association of papillomavirus with cancers of the head and neck

One hundred one squamous cell carcinomas (SCC) (study group) and 116 tissues without SCC or papilloma from matched anatomic sites (control group) were evaluated for the presence of human papillomavirus (HPV) DNA. Squamous cell carcinomas of the tongue (18%), tonsil (29%), and pharynx (13%) specifically harbored HPV-16-related sequences. In contrast, the nose, mouth, and esophagus proved completely negative. In the larynx, a low prevalence of HPV-11/16-related DNA was found in both cancers (5%) and control tissues (4%), suggesting that the associations lacked specificity. Our results indicate that anatomic site plays a role in determining the susceptibility to infection, and that the clinical entities with which HPV infections are associated include both subclinical infection with no history of papilloma, and malignant disease.     

Prospective screening for multiple tumors of the upper aerodigestive tract: a simple routine procedure

Detection of a second primary at the time of initial work-up of patients with head and neck cancers has an important bearing on management planning. In a 12-month period, a series of 105 patients with an index head and neck primary (skin malignancies excluded) of the squamous cell type were subjected to the following screening procedure. Under topical anesthesia, with the patient in the sitting position, a small-size flexible bronchofiberscope (Olympus B3R) was introduced through the nose. As the scope was advanced, the nasal cavity, nasopharynx, oropharynx, hypopharynx, and larynx were examined. The tracheobronchial tree was also examined if the chest x-ray showed abnormal findings. The endoscope was then passed down to examine the esophagus, with oxygen insufflated via the biopsy channel at a rate of 1 liter per minute. The index primary sites included the tongue (20), alveolus (7), floor of mouth (2), palate (4), buccal mucosa (3), hypopharynx (17), and larynx (52). There was little patient discomfort and no complication. Nine patients (8.5%) were found to have simultaneous primary tumors including two patients with triple primaries and one with quadruple lesions. The additional tumors were in the following sites: esophagus (6), tonsil (2), floor of mouth (1), nasopharynx (1), bronchus (1), palate (1), and oropharynx (1). The treatment plans were changed in all nine patients with the discovery of multiple tumors. It is concluded that the panendoscopic screening in the manner described is simple, safe, and convenient and the information is contributory to treatment planning.     

Evaluation and application of a broad-spectrum polymerase chain reaction assay for human papillomaviruses in the screening of squamous cell tumours of the head and neck

OBJECTIVE: The results of studies analysing the prevalence of human papillomaviruses (HPVs) in squamous cell carcinomas of the head and neck region often vary depending on the different molecular biological methods applied. Focusing on the histomorphological criteria of HPV infections, the percentage of HPV-positive cancers should be higher than has been found in most studies. The aim of this study was to increase the detection spectrum of HPV polymerase chain reaction (PCR) screening using degenerate consensus primers. MATERIAL AND METHODS: To increase the sensitivity of the assay to one copy of HPV DNA per cell, a two-step PCR was carried out. The products were directly sequenced by means of a cycle sequencing approach. Seventy biopsies from squamous cell carcinomas of the upper aerodigestive tract were screened using the primer system. RESULTS: According to the PCR results, 0/2 carcinomas of the floor of the mouth or tongue, 7/16 biopsies of oropharyngeal cancers, 3/13 hypopharyngeal cancers, 13/34 squamous cell carcinomas of the larynx, 4/4 biopsies from carcinomas of the nasal cavity or paranasal sinuses and 1/1 squamous cell carcinoma of the parotid gland were positive. CONCLUSION: This broad-spectrum PCR can effectively detect HPV in head and neck cancers.     

Evaluation and application of a broad-spectrum polymerase chain reaction assay for human papillomaviruses in the screening of squamous cell tumours of the head and neck

Objective—The results of studies analysing the prevalence of human papillomaviruses (HPVs) in squamous cell carcinomas of the head and neck region often vary depending on the different molecular biological methods applied. Focusing on the histomorphological criteria of HPV infections, the percentage of HPV-positive cancers should be higher than has been found in most studies. The aim of this study was to increase the detection spectrum of HPV polymerase chain reaction (PCR) screening using degenerate consensus primers. Material and Methods—To increase the sensitivity of the assay to one copy of HPV DNA per cell, a two-step PCR was carried out. The products were directly sequenced by means of a cycle sequencing approach. Seventy biopsies from squamous cell carcinomas of the upper aerodigestive tract were screened using the primer system. Results—According to the PCR results, 0/2 carcinomas of the floor of the mouth or tongue, 7/16 biopsies of oropharyngeal cancers, 3/13 hypopharyngeal cancers, 13/34 squamous cell carcinomas of the larynx, 4/4 biopsies from carcinomas of the nasal cavity or paranasal sinuses and 1/1 squamous cell carcinoma of the parotid gland were positive. Conclusions—This broad-spectrum PCR can effectively detect HPV in head and neck cancers.     

c-myc oncogene copy number in squamous carcinoma of the head and neck.

PURPOSE: Altered resident cellular genetic sequences (oncogenes) may result in malignant transformation, maintenance of tumor growth, and metastatic propensity. In this pilot study, we have elected to probe c-myc oncogene in evaluating specimens from human squamous cell carcinoma. MATERIALS AND METHODS: Samples were obtained from 24 patients with squamous cell carcinoma of the head and neck. The ratio of tumor DNA values to that of control DNA was used to estimate the c-myc copy number. RESULTS: Data from material obtained from eight patients was analyzed to the point of c-myc copy number. Tumors varied from stage II through IV. Five originated in the oral cavity and three in the larynx. Analysis of primary tumors demonstrated that two of eight had increased c-myc copy numbers. Histologically positive neck specimens were encountered in five of the study patients. Three demonstrated elevated c-myc copy numbers, two of which had had increased copy number at the primary site. CONCLUSION: This study confirms that c-myc amplification can be present in squamous cell carcinoma of the head and neck. c-myc Amplification may also be present in neck metastasis. Oncogene amplification in neck metastasis may indicate an increased metastatic propensity for individual tumor cells demonstrating c-myc amplification.     

Variability of genetic alterations in different sites of head and neck cancer

OBJECTIVE: Tumors arising from different sites of the head and neck area have different clinical behavior. However, most of the studies on genetic alterations in head and neck squamous cell carcinomas do not make a distinction between the sites within this area. The objective of this study is to compare the genetic alterations in three different sites of the head and neck (larynx, oropharynx, and hypopharynx). STUDY DESIGN: Prospective study. METHODS: Thirty-eight laryngeal, 29 oropharyngeal, and 37 hypopharyngeal carcinomas were studied. DNA from tumor and healthy tissue was evaluated for amplification of the oncogenes at 11q13 region (CCND1, FGF3, FGF4 and EMS1) and of the oncogenes MYC and ERBB1; for integration of the human papillomavirus (HPV) types 6b and 16; for loss of heterozygosity (LOH) at p53 and NAT2; and for the cellular DNA content. RESULTS: FGF3 and FGF4 showed a significantly higher frequency of amplification in hypopharyngeal tumors (P =.006 and P =.0002, respectively). CCND1 amplification had a nearly statistically significant (P =.072) higher frequency of amplification in hypopharyngeal tumors. Aneuploid tumors were found in a significantly lower proportion in the larynx (P =.03) compared with the other sites. For the other genetic alterations, no significant differences among the three sites were found. CONCLUSIONS: These results suggest that cancers originating from different sites in the head and neck may have different tumor biology. Therefore, they should be considered as different entities.     

DNA contents of individual cells from squamous cell carcinomas of the larynx and tongue.

In previous work a new method of histologic grading of malignancy in squamous cell carcinomas of the larynx was described. It was based on morphologic criteria representing the tumor cell population itself and the tumor host relationship. Nuclear polymorphism was revealed to be a very important factor for determining prognosis. This method of histologic grading of malignancy was in this work further analyzed by cytophotometric DNA determinations on individual cells obtained by scraping the surface of squamous cell carcinomas of the larynx and the tongue. Increased heterogenous cellular DNA distributions were found in cases with high histological malignancy grading. Cells in the carcinoma cases with a normal epithelial morphology had the same DNA content as that of control cells of normal buccal mucosa. Cells with higher DNA values all had malignant or suspicious morphology.     

人咽喉部良恶性肿瘤与乳头状瘤病毒关系的研究

采用免疫组化及ＤＮＡ斑点杂交技术检测人咽喉部乳头状瘤及鳞状细胞癌组织中人乳头状瘤病毒（ＨＰＶ）壳蛋白抗原及ＨＰＶ６、１１、１６、１８型ＤＮＡ。１１例乳头状瘤ＨＰＶ抗原与ＨＰＶ ＤＮＡ阳性率均为４５．５％。２２例鳞状细胞癌ＨＰＶ抗原阳性率２２．７％，ＨＰＶ ＤＮＡ阳性率２７．３％。乳头状瘤ＨＰＶ检出率与组织学检查的结果相符。提示咽喉部乳头状瘤及鳞状细胞癌的发生、发展与ＨＰＶ感染有关。     

Surgical treatment of squamous cell carcinoma of the base of the tongue

One hundred two patients with squamous cell carcinoma of the base of the tongue were treated by surgery. The grade of the tumor did not influence survival; however, survival was influenced by the stage of the disease and the operation selected. Operations that provided wider access decreased the incidence of local recurrence. Of the 102 patients, 23 percent required either total or partial laryngectomy. The survival curves of the 23 patients suggest that management of the larynx is a key factor in treating larger cancers. Overall, the five-year survival rate for the 102 patients was 42 percent. Surgery is the treatment of choice in squamous cell carcinoma of the base of the tongue.     

Changing patterns of failure of head and neck cancer

BACKGROUND: With the increased use of neoadjuvant therapy for advanced stage squamous cell carcinoma of the head and neck, we have observed an apparent change in the pattern of failure from predominantly locoregional sites to distant metastases. We reviewed the patterns of failure in cancers of the oral cavity, oropharynx, and larynx at our institution during the last decade. OBJECTIVE: To determine whether there has been a significant change in the patterns of recurrence from the historical locoregional failure to distant sites, and whether this change is associated with the increased use of multimodality therapy. METHODS: We reviewed cancer registry data on patients with squamous cell carcinoma of the head and neck diagnosed between January 1, 1988, and December 31, 1999. Sites included the oral cavity and oropharynx (including the tongue, floor of mouth, retromolar trigone, gingiva, tonsil, and lip) and larynx. RESULTS: Among 432 patients with squamous cell carcinoma of the head and neck, 280 (65%) had oral cavity and oropharyngeal cancers, and 152 (35%) had laryngeal cancers. Overall, 19% developed locoregional recurrence, and 8% developed distant failure. Although locoregional failure for oral cavity and oropharyngeal squamous cell carcinoma decreased from 26% to 16% from 1988-1993 to 1994-1999, distant failure increased significantly from 3% to 8%. During these periods, multimodality therapy was used in 62% of oral cavity and oropharyngeal cancers, and this rate remained essentially unchanged. For laryngeal cancer, locoregional and distant failure remained stable at 18% and 9%, respectively. In these laryngeal cancers, the use of multimodality therapy decreased from 60% to 46%, but this difference was not statistically significant (P =.43). CONCLUSIONS: Although locoregional control in oral cavity and oropharyngeal cancers has improved significantly with the use of multimodality therapy, the incidence of distant failure has doubled. In laryngeal squamous cell carcinoma, the patterns of failure have not changed significantly.     

Clinical and histopathological considerations for the diagnosis and treatment of verrucous carcinoma of the larynx

Verrucous carcinoma of the larynx is quite distinct from well-differentiated squamous cell carcinoma has a characteristic morphology and specific clinical behavior. It accounts for approximately 1%-2% of all laryngeal carcinomas. However, difficulties are encountered clinically in separating verrucous carcinomas from keratoses, verruca vulgaris and squamous cell carcinomas with a verrucous appearance. Between 1962 and 1982, 1504 patients with squamous cell carcinomas of the larynx were diagnosed and treated by staff physicians in the Department of Otolaryngology--Head and Neck Surgery, Washington University in St. Louis. Fifteen of these patients (1%) had verrucous carcinomas. These cases were studied in order to clarify any diagnostic problems, trace the biologic behavior of the tumors and report the results of the treatment used. Of these 15 patients, we were able to follow up 8. The latter had been treated only with surgery and none of them had received radiotherapy. One death occurred, but the patient died of other causes and remained free of cancer. Reports in the literature describe a high rate of recurrence of verrucous carcinomas following radiotherapy with occasional anaplastic transformation of tumors. No patients in our series developed a metastasis in the neck, indicating that neck dissection is not justified for patients with these neoplasms even though clinically enlarged neck nodes may be present.     

Transoral laser microresection for cancer of the larynx involving the anterior commissure

OBJECTIVE: To share the authors' experience of transoral laser microresection in the treatment of squamous cell carcinoma of the anterior commissure of the larynx. STUDY DESIGN: Retrospective review of 39 patients with squamous carcinoma involving the anterior commissure of the larynx, all managed with transoral laser microresection between September 1996 and December 2001. Twenty-three patients had pT1 or pT2 cancers. Sixteen patients had pT3 or pT4 disease, two with N2b neck metastases. Twenty patients had cancers exhibiting spread to the subglottis. Twenty-five patients had received prior treatment(s). METHODS: Specific data points entered into a portable database were retrospectively verified against the patient's electronic medical record. Sorting and analysis were completed in a desktop spreadsheet. We evaluated tumor recurrence, cause of death, tracheostomy, hospital length of stay, and voice quality. RESULTS: None of the 17 patients with early cancers (pT1 or pT2a) had local recurrence, and none received postoperative radiotherapy. Five of the 22 patients with intermediate or advanced disease (pT2b/pT3 or pT4 cancer) required second treatments for persistent or recurrent disease. Of these, two had small residual foci amenable to second-stage resection by laser. One patient developed a prelaryngeal soft tissue recurrence and retained his larynx after wide local excision and radiotherapy. Two patients (both previously irradiated) developed delayed recurrences requiring total laryngectomy. Four retreated patients remained alive without disease. One patient died of other causes. Voice remained no worse after transoral laser microresection in 19 patients. It was one level worse (on a scale of 0 to 5) for 16 patients. Eleven patients received temporary tracheotomies, some for airway, some for exposure. Twelve patients left the hospital on a same-day basis. The average hospital stay was 3.3 days. CONCLUSION: Transoral laser microsurgery is one of the options to be considered for the treatment of squamous cell cancer involving the anterior commissure of the larynx.     

Role of 18F-FDG PET in detecting primary site in the patient with primary unknown carcinoma

The aim of this study was to verify the effectiveness of positron emission tomography (PET) in detecting primary sites in carcinoma of unknown primary (CUP) patients. In this study, CUP represented a group of heterogeneous tumors that shared the clinical manifestation of metastatic carcinoma with no obvious primary site at the time of first diagnosis, which included clinical investigations, computed tomography, magnetic resonance imaging and panendoscopy. We reviewed the records of 24 patients with CUP between January 1995 and December 2009. The patients who demonstrated additional tracer uptake sites other than previously known metastatic lesions by PET scan were done direct biopsies for the sites of accumulation. Patients who had a negative PET scan or for whom the primary site could not be identified by direct biopsies underwent examination under anesthesia of the at-risk occult tumor sites. PET scan demonstrated focal accumulation suspicious for primary tumor in 12 (50.0%) of 24 patients: tonsil 5, nasopharynx 3, hypopharynx 1, tongue 1, larynx 1, and maxillary sinus 1. A subsequent biopsy of these sites revealed primary cancer in 9 (37.5%) of 24 patients: tonsil 5, nasopharynx 1, hypopharynx 1, tongue 1, and maxillary sinus 1. In the remaining three patients, no malignant cells were found by the biopsy of the accumulated area: nasopharynx 2, larynx 1. PET scans increase the yield of primary tumor by 37.5%. The sensitivity, specificity for PET scan were 80.8, 76.9%, respectively. PET scanning is useful in detecting primary cancer of CUP patients.     

Transfection of human nasopharyngeal epithelial cells with Epstein-Barr virus DNA

Transfection experiments using Epstein-Barr virus (EBV) DNA were carried out to examine whether three different epithelial cell lines (Ad-AH, D98, and KB) could be transfected and whether biologically active virus could be recovered. We found that the Ad-AH cells derived from the human nasopharynx were more transfectable, as determined by the synthesis of EBV-specific antigens. Concentrates of supernatant fluids obtained from transfected Ad-AH cells were capable of transforming human cord blood lymphocytes (HCBLs). However, no concentrates of supernatant fluids obtained from transfected epithelial cell lines outside the nasopharynx (D98 and KB) were capable of transforming HCBLs.     

Clinical and histopathological considerations for the diagnosis and treatment of verrucous carcinoma of the larynx

Summary Verrucous carcinoma of the larynx is quite distinct from well-differentiated squamous cell carcinoma and has a characteristic morphology and specific clinical behavior. It accounts for approximately l%–2% of all laryngeal carcinomas. However, difficulties are encountered clinically in separating verrucous carcinomas from keratoses, verruca vulgaris and squamous cell carcinomas with a verrucous appearance. Between 1962 and 1982, 1504 patients with squamous cell carcinomas of the larynx were diagnosed and treated by staff physicians in the Department of Otolaryngology — Head and Neck Surgery, Washington University in St. Louis. Fifteen of these patients (1%) had verrucous carcinomas. These cases were studied in order to clarify any diagnostic problems, trace the biologic behavior of the tumors and report the results of the treatment used. Of these 15 patients, we were able to follow up 8. The latter had been treated only with surgery and none of them had received radiotherapy. One death occurred, but the patient died of other causes and remained free of cancer. Reports in the literature describe a high rate of recurrence of verrucous carcinomas following radiotherapy with occasional anaplastic transformation of tumors. No patients in our series developed a metastasis in the neck, indicating that neck dissection is not justified for patients with these neoplasms even though clinically enlarged neck nodes may be present.On leave from the Department of Otorhinolaryngology, University of Kosovo, Prishtina, Yugoslavia     

喉外侵犯对声门上型喉癌部分喉切除术的影响

本文回顾分析了中国医学科学院肿瘤医院1979年至1990年168例声门上型喉癌行部分喉切除术患者的临床资料,研究了喉外侵犯对声门上型喉癌水平部分喉切除术的影响,发现舌根受侵切除部分舌根使水平部分喉切除术的拔管率由92.10％降为58.82％,梨状窝受侵者切除部分梨状窝使3/4部分喉切除术的拔管率由67.39％降为33.33％(P<0.05)。梨状窝内壁受侵组的生存率低于舌根受侵组(P<0.01)。因此,将声门上型喉癌梨状窝内壁受侵定为T_4病变更合适。结论:声门上型喉癌侵及舌根及梨状窝内壁时部分喉切除术后的拔管率降低,梨状窝内壁受侵时生存率也明显降低。     

Malignant Transformation of Recurrent Respiratory Papillomatosis Associated With Integrated Human Papillomavirus Type 11 DNA and Mutation of p53

Recurrent respiratory papillomatosis (RRP), usually confined to the nasopharynx, trachea, and larynx, occasionally can progress to extensive bronchopulmonary disease. Most cases of bronchopulmonary and laryngeal papillomatosis are cytologically benign and do not undergo malignant transformation; however, squamous cell carcinoma (SCC) can arise in RRP in the absence of known risk factors such as radiation and smoking. In this study, the authors investigated molecular genetic alterations occurring in a case of metastasizing SCC that arose in long-standing bronchopulmonary papillomatosis. Genomic DNA from tracheal papillomata, tracheobronchial papillomata, SCC of the lung, and a lymph node metastasis was extracted. The physical state of the human papillomavirus type 11 (HPV-11) DNA was investigated by two-dimensional gel electrophoresis. Molecular genetic alterations of the host genome were studied by direct sequencing of polymerase chain reaction-amplified gene fragments and restriction fragment length polymorphism (RFLP) analysis. Episomal and integrated forms of HPV-11 sequences were detected in histologically benign tumors, but only the integrated form of the viral DNA could be found in malignant tissue samples. Molecular genetic studies revealed that an allelic loss of the interferon-β gene (IFNβ-1) and an endogenous type of mutation of the p53 antioncogene were found only in the malignant lesions. Mutations were not observed in the ras, neu, or multiple tumor suppressor (MTS1/p16) genes in any specimens. The authors' data indicated that the p53 genetic mutation was associated with integration of HPV-11 in histologically malignant lesions. This association may promote a progressive genetic instability that can lead to the development and clonal expansion of malignant lesions in RRP.     

Transoral laser microsurgery for squamous cell carcinoma of the base of the tongue

OBJECTIVE: To determine the role of transoral laser microsurgery for base of tongue squamous cell carcinoma. DESIGN: Retrospective unicenter study of the oncologic and functional results of laser microsurgery of tongue base carcinoma performed between 1986 and 1997. SETTINGS: University hospital department. PATIENTS: We reviewed 48 previously untreated patients with base of tongue squamous cell carcinoma, who were treated with transoral laser microsurgery. Distribution of the T categories were T1, 2%; T2, 25%; T3, 15%; and T4, 58%; 94% belonged to the stages III and IVa. Selective neck dissection was performed in 43 patients; 23 patients underwent postoperative radiotherapy with or without simultaneous chemotherapy. MAIN OUTCOME MEASURES: Local control rate, recurrence-free and overall survival rates, mean performance status scale scores for normalcy of diet and understandability of speech. RESULTS: The Kaplan-Meier 5-year local control rate was 85%. There was no local recurrence in T1 and T2 lesions, but there was a 20% local recurrence rate in T3 and T4 tumors. Kaplan-Meier 5-year recurrence-free and overall survival rates were 73% and 52%, respectively. Mean performance status scale scores were 92% for normalcy of diet and 88% for understandability of speech. Twenty-one patients survived at least 5 years after treatment. They have a preserved larynx and live without tracheostoma or gastrostomy tube. CONCLUSIONS: Our concept of organ and function preserving laser microsurgery for selected patients with base of tongue cancer seems to be justified considering the achieved oncological and functional results. Final proof of the effectiveness of the new therapeutic concept presented herein requires well-designed prospective studies.