The tumour stroma of oral squamous cell carcinomas show increased vascularity compared with adjacent host tissue

For tumours to grow they must acquire an adequate blood supply, and the use of drugs to inhibit tumour vascularization is one promising approach to anti-cancer therapy. Clear information is therefore required on the vascular architecture of human tumours and animal tumour models used for testing anti-angiogenic therapies. Many previous studies on animal tumour models have shown that carcinomas are least vascular in their centres and that host tissues become more vascular with proximity to the tumour. However, we have previously found that many human colorectal carcinomas do not show this pattern. The present study on human oral squamous cell carcinomas (SCCs) again reveals significant differences. Paraffin sections from 24 SCCs were immunostained using the QBEnd-10 monoclonal antibody to demonstrate blood vessels, and these were quantified by interactive morphometry using a Kontron Videoplan system. In most carcinomas, viable tumour tissue was no less vascular in the tumour centre than in the tumour periphery. Although tumours are known to release angiogenic factors, viable tumour tissue was less vascular than adjacent host tissues. However, the tumour stroma, by itself, was more vascular than adjacent host tissues. Host tissue adjacent to tumour showed no obvious increase in vascular density with increasing proximity to the tumour edge, which suggests that tumour-released angiogenic factors are only effective over a short distance.     

Proliferating cell nuclear antigen (PCNA) expression in oral squamous cell carcinoma - an aid to conventional histological grading?

Decreased oxygen delivery and cellular hypoxia are major factors in the pathophysiology of shock. We studied the effects of 100% O2 at 0.1 and 0.3 MPa (1 and 3 atm abs) in severe hemorrhagic shock in awake, unrestrained rats. Shock was induced by withdrawing 50% of the total blood volume within 120 min. Blood pressure, heart rate, and the electroencephalogram (EEG) were recorded during the first 6 h of the protocol. The animals were observed for 7 days. The shock protocol resulted in 60 and 90% mortality after 1 day and at the end of 7 days, respectively. A single 90-min exposure to O2 at 0.1 and 0.3 MPa, which was started 30 min after bleeding, maintained mean arterial blood pressure at significantly higher values compared to untreated controls throughout the exposure period (P < 0.05). Oxygen therapy at both doses also improved the long-term survival rate and survival time significantly (P < 0.01). No clinical or EEG sign of CNS O2 toxicity was detected in O2-treated animals. Our results indicate that O2 given alone after severe bleeding exerts a beneficial effect on the long-term outcome of hemorrhagic shock in awake, unrestrained rats.     

Prognostic value of ERBB-1, VEGF, cyclin A, FOS, JUN and MYC in patients with squamous cell lung carcinomas

The A1 adenosine receptor (A1AR) contributes to the cytoprotective action of adenosine under conditions known to generate reactive oxygen species (ROS). Pharmacological manipulation of A1AR expression has been shown to modulate this cytoprotective role. In this study, we provide evidence that ROS generated could increase the expression of the A1AR and thereby offset the detrimental effects of ROS. Incubation of DDT1MF-2 smooth muscle cells with ROS-generating chemotherapeutic agents, such as cisplatin (2.5 microM) or H2O2 (10 microM), elicited an increase in A1AR expression within 24 hr. The induction by H2O2 was reduced by the ROS scavenger catalase but not superoxide dismutase. Inhibition of nuclear factor kappa B (NF kappa B) by pyrrolidine dithiocarbamate (200 microM), dexamethasone (100 nM), or genistein (1 microM) abrogated the cisplatin-mediated increase in A1AR. Cisplatin promoted rapid translocation of NF kappa B (but not AP-1) to the nucleus, as detected by electrophoretic mobility shift assays and by Western blotting. A putative NF kappa B sequence in the A1AR promoter effectively competed with labeled kappa B probe for binding in nuclear preparations derived from DDT1MF-2 cells. Transient transfection of DDT1MF-2 cells with the A1AR promoter coupled to firefly luciferase reporter gene led to cisplatin-inducible and pyrrolidine dithiocarbamate-sensitive luciferase activity, suggesting the presence of functional NF kappa B binding site(s) in the A1AR promoter sequence. Treatment of cells with (R)-phenylisopropyladenosine (1 microM), an agonist of the A1AR, reduced cisplatin-mediated lipid peroxidation, which was reversed after blockade of the A1AR. These data suggest that ROS can increase the expression of the A1AR by activating NF kappa B regulatory site(s) on this gene and thereby enhance the cytoprotective role of adenosine.     

Audit of clinical information and diagnoses supplied to the pathologist following biopsy of oral squamous cell carcinomas

Strict criteria for the biopsy of oral lesions suspected to be squamous cell carcinoma (SCCA) are difficult to find. Evaluation of the patient's history and clinical findings should help the clinician to accurately diagnose oral cancer. Our study attempts to compare and correlate cases that have the histopathologic diagnosis of oral SCCA with the data submitted by the clinician to the oral pathologist. We find the more information the clinician lists in the biopsy request form, the more likely the correct clinical diagnosis of oral SCCA is. We also show that when risk factors such as tobacco and alcohol use are mentioned by the clinician, the percentage of correct clinical diagnosis is increased.     

Prognostic value of initial ultrastructural changes in squamous cell metaplasia of junction cell papilloma of the bladder]

In 48 patients with a junction cell papilloma, in 65--with a junction cell and in 20--with nondifferentiated bladder cancer the electron-microscopic studying of the squamous cell metaplasia signs was conducted. Great prognostic importance of an early and overt signs of squamous cell metaplasia was established.     

Prognostic factors of cervical lymph node metastasis in head and neck squamous cell carcinoma

AIMS AND BACKGROUND: The metastatic spread of squamous cell carcinoma of the head and neck (SCCHN) to the cervical lymph nodes is a negative prognostic factor in terms of survival. We have used multivariate analysis to identify the possible prognostic significance of a number of clinical and pathological characteristics in relation to possible involvement of the cervical lymph nodes in a series of 396 patients. METHOD: 396 patients with SCCHN were studied. Variables regarding the patient, the carcinoma and histology were analysed by multivariate analysis using BMDP's PLR programme. RESULTS: Some variables appear to represent predisposing factors for tumor spread to the lymph nodes: tumor site (supraglottic larynx: P = 0.005; base of the tongue: P = 0.02; hypopharynx: P = 0.02), grading (P = 0.001), and a number of histological parameters (lower degree of histological differentiation: P = 0.001; vascular permeation: P = 0.04; perineural invasion: P < 0.05; prevalently plasmocytic infiltrate: P < 0.05). CONCLUSION: The identification of cases at risk for metastasis can be improved by the assessment of prognostic factors, with a consequent improvement in treatment strategies.     

Six primary squamous cell carcinomas of the head and neck

Estrogens have a beneficial effect on atherosclerosis and osteoporosis after menopause, but their exact mechanism of action is still unknown. The aim of the present study was to investigate the effects of estradiol and its metabolites catechol estrogens on arachidonic acid metabolism in vitro. Estradiol had no effect on arachidonic acid metabolism up to 33 microM in A23187-stimulated human whole blood. All catechol estrogens (2-hydroxyestradiol, 2-hydroxyestrone, 4-hydroxyestradiol and 4-hydroxyestrone) had similar kinds of actions on arachidonic acid metabolism, being over ten times more potent inhibitors of leukotriene synthesis (IC50 values 0.044-0.16 microM) than thromboxane (IC50 values 0.99-2.1 microM) and prostaglandin E2 synthesis (IC50 values 0.84-5.5 microM). It is suggested that some of the protective actions of estrogens--e.g., on atherosclerosis and osteoporosis--may be related to the inhibition of leukotriene synthesis by catechol estrogens.     

Comparison of DNA copy numbers in original oral squamous cell carcinomas and corresponding cell lines by comparative genomic hybridization

We analyzed regional DNA copy numbers in 4 oral squamous cell carcinomas (SCCs) by using comparative genomic hybridization, and compared them with those in cell lines derived from the SCCs. In the original tumors, DNA copy number increases were observed on chromosomes 5p (4/4 cases), 8q (4/4), 20p (3/4), 3q (2/4), 5q (2/4), 7p (2/4), 7q (2/4), 11p (2/4), 11q (2/4) and 13q (2/ 4). Although most of these changes have been described previously for SCC tumors in the head and neck, the incidence of increases in 8q and 20p was much higher in the present study; this may be important in relation to cell line establishment, since 8q contains e-myc, which is involved in immortalization. No common chromosomal region with DNA copy number decreases was observed, except for 18q (2/4). When the original tumors and the cell lines were compared, their profiles were essentially similar with one exception. Further, there was no region that commonly changed in the cell lines, but not in the original tumors, suggesting that the DNA copy number changes observed in the cell lines mostly represent those of the original tumors.     

Prognosis value of the expression of Ki-67 for squamous cell carcinoma of the oral cavity

A single dose of either cyclosporin-A (CsA) or lobenzarit (CCA) given with an arthrogenic adjuvant completely prevented expression of experimental adjuvant arthritis in rats. The aim of this study was to understand how these drugs prevented the arthritis expression by studying the popliteal lymph nodes draining the arthritic joints at various times after adjuvant injection. Neither drug affected the proliferation in popliteal lymph nodes at the time arthritis was normally expressed, however, there was a marked change in the types of cells present. Immunofluorescence assays showed a reduction in the proportion of CD4+ cells, while the proportion of B-lymphocytes was almost doubled. This coincided with a marked elevation in the ability of these cells to produce interleukin (IL)-6. At the same time production of other cytokines (IL-2, tumour necrosis factor (TNF) and interferon (IFN)-gamma) was not greatly affected. However, one day after adjuvant injection IL-2 and IFN-gamma production was reduced. In vitro experiments showed that IL-6 production by lymphoid cells was relatively unaffected by CsA and CCA but IL-2, TNF and IFN-gamma were suppressed by CsA. The results indicate that CsA and CCA may modify the response to the arthritic adjuvant by specifically inhibiting IL-2, TNF and IFN-gamma production at the time of adjuvant injection. The lack of inhibition of IL-6 by these drugs reveals it may not play a key role in the initiation of this model of chronic inflammation.     

Prognostic factors and management of carcinomas of the gallbladder and extrahepatic bile ducts

Cancers of the biliary tract are uncommon but aggressive malignancies that pose difficult problems in diagnosis and management. Long-term survival with these cancers is limited by their propensity for local invasion, so that pathologic stage becomes a major prognostic factor, and by their ability to cause biliary obstruction and sepsis and interfere with hepatic function. In selected patients, surgical resection offers the possibility of cure, but effective palliation is often the principal goal of treatment. Radiologic and endoscopic modalities thus often play a major role in patient management.     

Fibrin deposition in squamous cell carcinomas of the larynx and hypopharynx

Extravascular fibrin deposition is frequently observed within and around neoplastic tissue and has been implicated in various aspects of tumor growth. The distribution of fibrin deposits was investigated in squamous cell carcinomas representing different stages of tumor progression of the larynx (n = 25) and hypopharynx (n = 9) by immunofluorescent techniques. Double and treble labelings were used to detect fibrinogen and fibrin in combination with marker antigens for tumor cells (cytokeratin), endothelial cells (von Willebrand factor), macrophages (recognized by KiM7), as well as factor XIII subunit A (FXIIIA) and tenascin (an embryonic extracellular matrix protein newly expressed during tumorigenesis). All tissue samples showed specific staining for fibrinogen/fibrin. Fibrin deposition was localized almost exclusively in the connective tissue compartment of tumors with characteristic accumulation at the interface of connective tissue and the tumorous parenchyma. In certain tumor samples showing highly invasive characteristics, fibrin deposits were observed in close association with tumor blood vessels in the tumor cell nodules. The overlapping reactions with polyclonal antibody to fibrinogen/fibrin and monoclonal antibody to fibrin indicate the activation of the coagulation cascade resulting in in situ thrombin activation and fibrin formation. Fibrin was crosslinked and stabilized by FXIIIA as revealed by urea insolubility test. Accumulation of phagocytozing macrophages detected by Ki M7 monoclonal antibody could be seen in areas of fibrin deposition. The blood coagulation factor XIIIA was detected in and around the cells labeled with Ki M7 antibody. Tenascin and fibrin deposits were found in the same localization in the tumor stroma and in association with tumor blood vessels within the tumor cell nodules. Neither fibrin nor tenascin were detected in the histologically normal tissue adjacent to tumors. The close association between fibrin deposits and macrophage accumulation strongly suggests the active participation of tumor-associated macrophages in the formation of stabilized intratumoral fibrin that facilitates tumor matrix generation and tumor angiogenesis.     

Overexpression of scatter factor and its receptor (c-met) in oral squamous cell carcinoma

HYPOTHESIS: Scatter factor (SF) is a pleiotropic growth factor that recently has been shown to induce epithelial cell proliferation, random motility, and invasion via interaction with its receptor, a tyrosine kinase encoded by the c-met proto-oncogene. Studies involving a variety of solid tumors have suggested that overexpression of the SF/c-met ligand-receptor pair is associated with the acquisition of a malignant phenotype. We hypothesize that SF and c-met are overexpressed in epithelial malignancies of the head and neck including squamous cell carcinoma (SCC) of the oral cavity. STUDY DESIGN: Immunohistochemical staining of randomly selected normal, dysplastic, and malignant oral tissues. METHODS: Formalin-fixed, paraffin-embedded tissues were obtained from the Department of Oral Pathology at Shands Hospital (University of Florida), Gainesville, Florida. Examples of mild dysplasia, severe dysplasia, well-differentiated SCC, moderately differentiated SCC, and poorly differentiated SCC were randomly selected from the dictated reports of one of two staff oral pathologists. Histologically normal margins of each specimen served as normal controls. The tissues were immunohistochemically stained using commercially available antibodies against SF and c-met. Appropriate negative controls were run with each batch to ensure staining specificity. Evaluation of staining intensity was carried out using a computerized image analysis system. A one-way analysis of variance (ANOVA) with pairwise multiple-comparison procedures (Fisher method) was used to analyze the data. RESULTS: Statistically significant differences (P < .0001) in the intensity of staining were noted between the malignant and normal and the malignant and dysplastic tissues for both SF and c-met. No differences were appreciated when staining of normal and dysplastic sections of the SF-stained tissue were compared. CONCLUSIONS: The results suggest that the SF/c-met ligand-receptor pair is overexpressed in SCC of the oral cavity.     

The prognostic impact of metastatic pattern of lymph nodes in patients with oral and oropharyngeal squamous cell carcinomas

In patients with squamous cell carcinomas of the oral cavity and the oropharynx the presence or absence of nodal metastases still is the most important predictive factor. The discriminative significance of extracapsular spread and the influence of features of the primary tumor-such as size and depth of invasion-on metastatic pattern, treatment failure and survival were evaluated. Five-year postoperative follow-ups of 115 consecutively treated patients were studied retrospectively concerning the incidence of distant metastases, local and regional recurrences and the 5-year survival rate. Maximum depth of invasion of the primary tumor and lymph node metastases were evaluated on the basis of histological patterns, and patients were grouped according to their histological diagnosis. The T4 category has a plain discriminative influence on the incidence of distant metastases, recurrent tumors and survival rate in contrast to the other T sizes. The classification N0, intranodal growth and extranodal growth of lymph node metastases resulted in a 5-year survival rate of 67, 59 and 31%. According to the classification, 84, 87 and 59% were without nodal recurrence after 5 years, and 79, 82 and 46% without distant metastases. Size and depth of invasion of the primary tumor are not connected significantly with the occurrence of extracapsular spread. The status of the lymph nodes in squamous cell carcinomas of the oral cavity and the oropharynx metastases and in particular the capsular rupture has the most significant prognostic influence. The histological feature of extracapsular spread could distinguish reproducibly high risk patients with squamous cell carcinomas of the oral cavity and the oropharynx.     

Prognostic value of Ia-antigen, fibronectin, laminin and type IV collagen in pulmonary squamous cell carcinoma

Presence of absence of four immunohistological markers were investigated in fifty-four cases of pulmonary squamous cell carcinoma. Ia-antigen (HLA-DR), fibronectin (Fn), laminin (La), and type IV collagen were present in 9 cases (16.7%), 13 (24.0), 24 (44.4) and 12 (22.2) respectively. Three markers were present in 6 cases, two 13, one in 14 and none in 21. Higher positive rate of Fn among N (-) group in TNM classification was statistically significant when compared with that of N (+) group. In 13 of 14 cancer death cases, less than one markers were present, while all the cases in which more than two markers were present were alive during the follow up periods. The author concludes that Fn by itself is a useful prognostic indicator and when combined with other three markers, it can predict accurate prognosis.     

Prognostic factors for loco-regional control and outcome of re-irradiation for patients with poorly-differentiated squamous cell carcinoma of the nasopharynx

Retrospective analysis was performed to evaluate the prognostic factors for loco-regional control and the results of re-irradiation for 28 patients with recurrent, poorly-differentiated squamous cell carcinoma (PDSCC) of the nasopharynx. Twenty-four of them received re-irradiation. Local, local plus regional and regional recurrences were observed in 19, five and four patients, respectively. Except for three patients, all had Stage IV disease at the initial diagnosis. The only parameters influencing loco-regional recurrence were T and N stage categories. The median latent period from initial treatment to recurrence was 18.5 (range, 2-100) months. There was no difference in latent period by first recurrence site, although recurrent tumors confined to the nasopharynx or those only regionally developed had a longer latent period. Only four patients developed secondary distant bone metastases with a median latency of three months from loco-regional relapse. The patients with local recurrent tumors confined to the nasopharynx, and those with regional recurrences only, could be salvaged by re-irradiation, with five-year survival rates of 44 and 100%, respectively. Five of 28 patients (18%) developed severe chronic radiation sequelae: cerebrospinal complications in four patients, bilateral neck fibrosis in one. We conclude that recurrent PDSCC of the nasopharynx can be controlled by re-irradiation with some success. Radiation therapy techniques must, however, be carefully planned in order to avoid the severe late post re-irradiation sequelae. For patients with advanced non-curable local recurrences, palliative care should be recommended instead of agressive re-irradiation.     

Clinicopathological study on patterns of cervical lymph node metastases from squamous cell carcinomas (SCC) of the head and neck

In order to investigate the patterns of cervical lymph node metastases from head and neck SCC, serial sections were performed on 384 radical neck dissection (RND) specimens. Positive lymph node was found in 60.4% RNDs. The cervical lymph node spread from SCC in the head and neck regions including oral cavity, oropharynx, hypopharynx and larynx has some predictable patterns, i.e., for primary SCC of the oral cavity, the majority of cervical lymph node metastases were clustered at levels I, II and III; and for primary carcinoma of the oropharynx, hypopharynx and larynx, a majority of node metastases were located at levels II, III and IV. The positive lymph nodes mainly distributed at only one level or consecutive levels. The rates of pathologically positive lymph node and extranodal spread grew with the increase of the clinical N-staging. It is suggested that supraomohyoid neck dissection (levels I, II and III) is particularly applicable to carcinomas of the oral cavity, and lateral neck dissction (levels II, III and IV) is applicable to carcinomas of the oropharynx, hypopharynx and larynx in patients with limited (N0 and N1) neck nodules, but for patients with N2 and N3 nodules, RND is neccessary to eradicate the nodal metastases. Moreover, the postoperative radiotherapy is indispensable for ruling out the occult cervical lymph node metastaese in selective neck dissection.     

Erythrophagocytic tumour cells in melanoma and squamous cell carcinoma of the skin

BACKGROUND: Radial keratotomy may induce late hyperopic shift. We present data on 140 consecutive eyes with a follow-up of up to 3 years that underwent radial keratotomy with the RK suction bridge. METHODS: We conducted a retrospective study of 140 consecutive eyes that had radial keratotomy between 1987 and 1994. Mean preoperative spherical equivalent was -5.21 D (range -2.00 to -9.75 D). All operations were performed by one surgeon (JHK) with the RK suction bridge. A suction ring maintaining physiological intraocular pressure immobilized the eye and left a peripheral rim of uncut cornea. The ring incorporated an eccentric bridge that guided the radial keratotomy knife. The knife setting was 90% of the central corneal thickness, measured by pachymetry. Spherical equivalent refraction and spectacle corrected visual acuity were measured at 1 week, 1, 3, 6 months, 1 year, and 3 years after radial keratotomy. RESULTS: The mean preoperative spherical equivalent refraction of -5.21 D dropped to -0.43 D at 1 week (n = 136), -0.71D at 1 month (n = 120), -0.85 D at 3 months (n = 95), -0.74 D at 6 months (n = 73), -0.77 D at 12 months (n = 79), and -0.85 D at 3 years (n = 67). Compared to 1 month spherical equivalent, at 3 years three eyes (4.4%) had moved > = or 1.00 D toward hyperopia. One eye (1.4%) shifted by 1.25 D. Paired t-tests of mean spherical equivalent refraction did not reveal significant shifts toward hyperopia. Mean preoperative spectacle-corrected visual acuity was slightly diminished at 1 week and was equal or better thereafter. CONCLUSIONS: Our 3-year data suggest that a late hyperopic shift following radial keratotomy may be prevented if an intact peripheral rim is maintained and cutting depth does not exceed 90% of the lowest corneal thickness.     

Detection of human papillomavirus DNA sequences in oral squamous cell carcinomas and their relation to p53 and proliferating cell nuclear antigen expression

BACKGROUND: The etiology of oral squamous cell carcinoma (SCC) is still obscure. Since human papillomavirus (HPV) DNAs are associated with carcinoma of the uterine cervix, carcinomas of the oral cavity were investigated to ascertain if these viruses are present in squamous carcinomas of this anatomic site. METHODS: Seventy-seven oral mucosal SCCs were examined for the presence of HPV DNAs by polymerase chain reaction and dot blot hybridization. Immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and p53 was performed and single strand conformation polymorphism analysis for p53 was undertaken. In situ hybridization detection of HPV-16 DNA also was performed. RESULTS: Human papillomavirus-16 DNA was detected in 23 cases of oral SCC and both HPV-16 and HPV-18 DNA were detected in one case of tongue SCC. Human papillomavirus DNAs were detected of 11 of 33 tongue, 4 of 15 gingival, 2 of 4 palate, 2 of 5 buccal mucosa, 3 of 7 maxillary sinus, and 2 of 11 the floor of the mouth SCCs. None were detected in SCCs of the retromolar region (0/2). Immunohistochemical examination for p53 was performed in 26 cases of oral SCC and the accumulation of p53 protein was observed in 6 cases (i.e., in 4 of 17 HPV DNA-negative cases and in 2 of 9 HPV DNA-positive cases). Single strand conformation polymorphism analysis confirmed gene mutations in all 6 cases. Human papillomavirus-16 DNA was predominantly identified in cancer cells that showed a morphologic resemblance to basal cells and its hybridized signal in keratinized cells was reduced by in situ hybridization detection. Immunohistochemical detection of PCNA revealed its cooccurrence with HPV-16 DNA in cancer cells. CONCLUSIONS: These results suggest that HPV-16 DNA sequences may have the capability to maintain the proliferative state of epithelial cells, and may contribute to the production of malignant phenotypes.