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PubMed1975

Microfilaments in human epithelial cancer cells.

The occurence, distribution, and ultrastructural morphology of microfilaments in malignant epithelial cells of invasive squamous cell carcinoma of human oral cavity were studied by electron microscopy. The findings are compared with those in malignant oral epithelial cells of carcinoma-in-situ. In the malignant cells of invasive carcinoma, microfilaments 50-70 A in diameter are prominent in the cortical cytoplasm of the lateral and basal cell surfaces, adjacent and parallel to the plasma membrane, and extending into cell processes and microvillous extensions. Additional microfilaments are found to run from the peripheral cytoplasm to the perinuclear region. The microfilaments are aggregated into bundles aligned parallel to the long axis of the cell and display foci of increased electron density. They also tend to be aggregated into complex polygonal arrays. These microfilaments are similar in organization, concentration and ultrastructural architecture to those of various other nonmuscle cells, where they are thought to be capable of contraction and associated with cell motility. The presence of a microfilament system believed to be associated with contractile and motile cell processes may be an important characteristic of malignant cells of invasive tumors. The lack of abundant organized microfilaments in malignant cells in the absence of tumor invasion, and the presence of a prominent microfilament system in cells of invasive tumors, suggest that the microfilaments are related to the invasive properties of malignant tumor cells.

Basement Membrane↗
PubMed1987

In situ DNA hybridization analysis of human papillomavirus (HPV) sequences in benign oral mucosal lesions.

A series of 144 surgically treated benign oral mucosal lesions were analysed using an in situ DNA hybridization technique with 35S-labeled human papillomavirus (HPV) DNA probes to demonstrate the DNA of HPV types 6, 11, 13, and 16, in routinely processed, paraffin-embedded biopsy specimens. These lesions and an additional 62 benign oral mucosal biopsy specimens (total, 206 specimens) were also assessed by the indirect immunoperoxidase (IP-PAP) technique to detect the expression of HPV structural proteins (viral antigens). A total of 54/206 (26.2%) lesions were observed to express HPV antigens, being found in 45/92 (48.9%) of the squamous cell papillomas/condylomas, in 1/54 fibrous hyperplasias, in 1/8 true fibromas, and in 7/8 (87.5%) of the focal epithelial hyperplasia (FEH) lesions. Of the HPV DNA-positive lesions, 15/45 (33.3%) expressed HPV antigens, the expression not being related to any particular HPV type. HPV DNA sequences were found in 45/144 (31.3%) of the lesions. HPV DNA was present with the highest frequency in FEH (83.3%), followed by the papilloma/condyloma group (33.8%), papillary hyperplasia (28.6%), fibrous hyperplasia (24.4%), and true fibromas (14.3%). The most frequent HPV type was HPV 11, representing 37.8% of the DNA-positive lesions. HPV 13 DNA, previously regarded as specific to FEH, was disclosed as a single HPV type in seven cases, and as a double infection by HPV 11 and 13 in an additional three cases, including all five morphologically distinct entities. Noteworthy is the discovery of the high-risk HPV type 16 DNA in 17.8% of the DNA-positive lesions, four papilloma/condyloma lesions, three fibrous hyperplasias, and one FEH.(ABSTRACT TRUNCATED AT 250 WORDS)

Condylomata Acuminata↗
PubMed1977

[Pathogenesis and clinical observations of papillomatous diseases of the skin and of the mucous membranes (author's transl)].

In the certain skin-warts, condylomata acuminata, oral-, laryngeal- and bronchial papillomas a morphologically identic virus is often seen. This virus belongs to the papova-virus-group and causes papillomas: Independant of their localization their clinical behaviour is very similar. Using cytophotometric methods we tried to find out if there are any signs which allow to reveal malignancy in continous reappearing papillomas.

Adult↗
PubMed1988

The relationship between epithelial Ia expression and the inflammatory cell infiltrate during experimental oral carcinogenesis.

The development of oral epithelial expression of Ia antigens and its relationship to the presence of IL-2r+ (CD25+) cells was investigated in rats treated with the water soluble carcinogen 4-nitroquinoline-N-oxide (4NQO). Acetone fixed frozen sections of the palate and tongue were stained using an indirect immunoperoxidase technique and monoclonal antibodies to rat Ia (I-A & I-E) and IL-2 receptor. After 4 weeks 4NQO treatment all rats expressed oral epithelial Ia but thereafter (2-9 months) expression was present in only 20-40% of animals. Epithelial expression of Ia by histologically normal, dysplastic and neoplastic epithelium was always associated with the presence of an underlying inflammatory cell infiltrate containing CD25+ cells. Overall there were significantly more CD25+ cells in tissue specimens containing Ia+ epithelium compared with Ia- epithelium. Furthermore, during the first 4 weeks of carcinogen treatment, a significant positive correlation was found between the CD25+ cell density and occurrence of focal epithelial Ia expression. These results, together with analysis of the T cell, NK cell, macrophage and B cell content of the infiltrates induced by 4NQO, suggest that the CD25+ cells represent activated T cells. Thus, our results in this experimental model are consistent with the idea that epithelial expression of Ia is the result of production of IFN-gamma by locally activated T cells.

4-Nitroquinoline-1-oxide↗
PubMed1980

[Electroacupuncture inhibition of nociceptive responses in the caudal trigeminal nucleus].

Acute experiments on cats showed that auricular electroacupuncture (AEAP)) produces significant inhibition of evoked potentials (EP) in the caudal trigeminal nucleus which occur under single stimulation of the tooth pulp. The inhibition of EP lasts for 60--80 min. Then gradual (over 30--40 min) recovery of EP is recorded. Acupuncture stimulation decreases insignificantly the amplitude of EP caused by irritation of mouth mucosa. This fact indicates that AEAP inhibits nociceptive responses but does not affect mediation of signals in other modalities. It was established that reacing of AEAP analgetic effect does not require that the classical scheme of acupuncture point localization be strictly followed.

Acupuncture Therapy↗
PubMed1993

Expression of lectin-specific cellular glycoconjugates during oral carcinogenesis.

The binding pattern of two lectins, concanavalin A (ConA) and peanut agglutin (PNA), in various phases of tumour progression in the oral epithelium was studied. These included non-dysplastic, dysplastic and neoplastic lesions as well as normal tissue. ConA and PNA showed intense staining in the basement membrane of all types of lesions. Little difference was observed in the staining patterns between different stages of oral carcinogenesis, either with ConA or PNA. ConA showed mild cytoplasmic and membrane staining in all types of lesions while PNA showed moderate to intense staining in both the cytoplasm and membrane of lower-layer cells in all histological groups. The present study therefore shows that these lectins have limited value in the elucidation of oral carcinogenesis and are of insignificant diagnostic value.

Carcinoma↗
PubMed1984

Kaposi's sarcoma. Gastrointestinal involvement correlation with skin findings and immunologic function.

We have evaluated 19 homosexual/bisexual male patients with biopsy-proven Kaposi's sarcoma (KS) of the skin in order to define the extent of gastrointestinal involvement and determine its correlation with oral mucosal disease, skin findings, and immunologic function. Nearly half the patients had oral mucosal lesions. In patients with oral mucosal lesions, 75% had gastrointestinal lesions. Some gastrointestinal involvement during the period of observation was present in 10 of the 19 patients. Involvement of the upper gastrointestinal tract was more common than colonic involvement: esophagus 1, stomach 8, duodenum 3, and colon 6. Significant immunosuppression was observed in these patients, measured in vitro by natural killer (NK) assay, and lymphocyte proliferation response to mitogens.

Adult↗
PubMed1996

Keratin antigen retrieval in oral mucosal biopsies using microwave processing.

In immunohistochemistry, it is well known that the majority of monoclonal antibodies to keratins work best on fresh frozen tissue specimens, yet in clinical practice most biopsies are routinely fixed in formaldehyde. This seriously limits the range of keratins that can be reliably assessed in retrospective studies (particularly where only rare archival material exists) and where subtle changes during tissue differentiation may be important. Antigen retrieval using exposure to microwave radiation is one technique that has been applied successfully to other tumour markers (e.g., p53). However, few papers have used this method when immunolabelling for keratins, in spite of the widespread use of antikeratin antibodies as markers of differentiation. The effect of keratin antigen retrieval using microwave processing was assessed on a range of oral mucosal biopsies, since the oral cavity displays a wide range of keratins. A panel of six well characterized antibodies was chosen: LP34 (Ck1, 5, 6, 18), LH1 (Ck10), LL025 (Ck16), A53 BA2 (Ck19), AE8 (Ck13), and E3 (Ck17). For each specimen, one piece was stored in liquid nitrogen and another piece fixed in formalin. Tissue sections were cut from each and, using the peroxidase avidin biotin technique, keratin expression was recorded for a frozen section, a dewaxed section, and a microwave-heated dewaxed section. Although overall there was a 25% improvement in identification of keratins after microwaving, some antibodies performed better than others. Given that keratins have been shown to be of value in tumour diagnosis, this study suggests that microwave processing of archival material can be a valuable adjunct to such analysis.

Antibodies↗
PubMed1988

Establishment of two cell lines from hamster buccal pouch tumors induced by topical 7,12-dimethylbenz(a)anthracene and topical DMBA in conjunction with herpes simplex virus infection.

Two cell lines designated HBPC-1 and HBPC-2 have been established from hamster buccal pouch tumors induced by topical 7,12-dimethylbenz(a)anthracene (DMBA) and DMBA in conjunction with type 1 herpes simplex virus infection, respectively. The cells are epithelial in morphology, have a doubling time of approximately 18 h, and require bovine serum for optimal growth. The karyotype is aneuploid, with several marker chromosomes, and the cells produce squamous cell carcinomas when transplanted into normal hamster pouch tissues.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1991

Three cell lines from hamster buccal pouch tumors induced by topical 7,12-dimethylbenz(a)anthracene, alone or in conjunction with herpes simplex virus inoculation.

Three squamous carcinoma cell lines HBPC-1, HBPC-2, and HBPC-3 were established from hamster buccal pouch tumors induced by topical 7,12-dimethylbenz(a)anthracene (DMBA) treatment alone, topical DMBA treatment in conjunction with type 1 herpes simplex virus (HSV-1) inoculation, and topical DMBA application in combination with type 2 HSV (HSV-2) inoculation, respectively. The cells were epithelial in morphology, had a doubling time of approximately 18 h, and required bovine serum for optimal growth. They demonstrated an in vitro anchorage-independent growth and produced squamous cell carcinomas when transplanted into normal hamster pouch submucosa. The carcinoma cell lines equally expressed cellular hst, src, abl, and raf proto-oncogenes that were not expressed in the normal hamster pouch epithelial cells. An equal amount of fos gene expression was noticed in the normal pouch epithelial cells, HBPC-1 and HBPC-3, but the HBPC-2 expressed less fos poly(A+)RNA than the other cell lines. The myc proto-oncogene was also expressed both in the normal pouch epithelial cells and in the cancer cell lines. However, the size and number of expressed myc poly(A+)RNA in the normal cells and cancer cell lines differed. Although the normal cells and HBPC-1 expressed a single myc transcript, 1.7-kilobase (kb) and 2.3-kb, respectively, both HBPC-2 and HBPC-3 expressed two myc poly(A+)RNAs, 1.7-kb and 2.3-kb.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1984

Hemidesmosomal dimensions and frequency in experimental oral carcinogenesis: a stereological investigation.

Connective tissue invasion by carcinomas results in a variety of morphological changes at the epithelial-connective tissue junction. In this report, hemidesmosomal size (-delta) and frequency (Ns) were quantified using established stereological methods. Following treatment of hamster cheek pouches with the chemical carcinogen DMBA, lesions defined as hyperplasias, dysplasias and carcinomas were evaluated using electron microscopical techniques. Untreated pouches served as controls. Stereological intersection counting procedures were used to quantify hemidesmosomes at the epithelial-connective tissue junction. During carcinogenesis, hemidesmosomal diameter decreased progressively, although the reductions were not statistically significant. Hemidesmosomal frequency also decreased progressively between control and carcinoma groups. Normal basal plasma membranes possessed approximately 8 hemidesmosomes/micron 2 whereas values were reduced to approximately 3/micron 2 in the carcinoma group. The reductions in hemidesmosomal frequency during carcinogenesis may reflect an increased motility of basal epithelial cells and a decreased adherence of basal cells to the adjacent lamina propria. The specificity of this alteration in the pathogenesis of malignancy can be readily determined by the application of stereological methods to various neoplastic and non-neoplastic lesions.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1984

Changes occurring in plasma membranes and intercellular junctions during the process of carcinogenesis and in squamous cell carcinoma.

A 0.5% mineral-oil solution of 9.10-dimethyl-1.2-benzanthracene (DMBA) was applied to artificial cecal pouches in the lower lips of rats. Ultrastructural studies were made of plasma membranes and intercellular junctions during the process of malignant transformation in the oral mucosal epithelium and after squamous cell carcinoma had been induced by the carcinogen. After the administration of DMBA, the inner leaflet of the membranes where the microfilaments are attached showed high electron density and intramembranous particles on the P-face of basal cells decreased to about half that of controls. However, on the E-face the number of intramembranous particles increased by approximately 10% compared with controls. Though the normal size range for intramembranous particles was 9-12 nm, the administration of DMBA caused aggregations of from three to six particles on the P-face. In squamous cell carcinomas, only the outer leaflet of the membranes showed high electron density; the number of intramembranous particles was 30% higher on the P-face and approximately three times higher on the E-face compared with controls and the morphology of the intramembranous particles, which formed irregular aggregates of from five to 20 particles, was specific. In animals treated with DMBA, the number of gap junctions decreased by between 50% and 70%, although no structural changes occurred. In squamous cell carcinomas, the area of gap junctions was about 50% lower and the number of gap junctions about 40% lower than in controls. Changes in the number and area of desmosomes were similar to those of gap junctions both in the DMBA-treated animals and in squamous cell carcinomas.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1997

DNA analysis of squamous cell carcinoma in the oral cavity by image cytometry and its rating in the assessment of tumor prognosis.

Image cytometric measurements on the DNA content of squamous cell carcinoma of the oral cavity were carried out to evaluate its relationship with clinicopathological features. In the prospective study 52 patients with operable oral cavity carcinoma in stage T1-4 and N0-3 were studied. The tumors were classified according to the TNM classification (UICC 1987) and graded histopathologically. A positive correlation between tumor size and ploidy status was observed. There was a significant correlation between ploidy status and histologically confirmed cervical lymph node metastases (P < 0.03). A relationship between histopathological grading and ploidy status was not found. The cumulative 5-year survival rate was significantly higher (P < 0.03) among patients with diploid DNA findings than among those with nondiploid findings. When assessing the prognosis of squamous cell carcinoma, DNA cytometry analysis should be used as an additional method. Particularly the primarily existing, significantly higher N stage, a higher frequency of metastases, and a significantly lower survival rate with nondiploid tumors underline the importance of this examination method.

Adult↗
PubMed1997

[Expression of proliferation markers PCNA and MIB 1 in verrucous squamous epithelial carcinoma of the oral cavity].

Expression of proliferation markers PCNA and MIB 1 was evaluated in 17 verrucous carcinomas and in five cases of normal squamous epithelium of the oral cavity. Immunohistochemistry revealed no significant differences in the proliferative activities. Highest expression was seen in the basal zones. Proliferative activity in more than 26% of cells is no longer compatible with verrucous carcinoma, but favours the diagnosis of squamous cell carcinoma.

Aged↗
PubMed2001

Micro-array chip analysis of carbonyl-metabolising enzymes in normal, immortalised and malignant human oral keratinocytes.

Enzymes involved in various protective and metabolic processes of carbonyl compounds were analysed utilising a micro-array method in a three-stage in vitro model for oral carcinogenesis involving cultured normal, immortalised and malignant human oral keratinocytes. A complete transcript profiling of identified carbonyl-metabolising enzymes belonging to the ADH, ALDH, SDR and AKR families is presented. Expression of 17 transcripts was detected in normal, 14 in immortalized and 19 in malignant keratinocytes of a total of 12,500 genes spotted on the micro-array chip. For the detected transcripts, about half were changed by cell transformation, and for the various enzyme families, differences in expression patterns were observed. The detected AKR transcripts displayed a conserved pattern of expression, indicating a requirement for the keratinocyte phenotype, while most of the detected SDRs displayed changed expression at the various stages of malignancy. The importance of multiple experiments in using a microarray technique for reliable results is underlined and, finally, the strength of the method in detecting co-expressed enzymes in metabolic pathways is exemplified by the detection of the formaldehyde-scavenging pathway enzymes and the polyol pathway enzymes.

Alcohol Dehydrogenase↗
PubMed2001

Myofascial flap without skin for intra-oral reconstruction. 1: Animal studies.

BACKGROUND: Although myofascial flaps are already used clinically in intra-oral reconstruction, the source and process of epithelialization remain unclear. METHODS: The process of epithelialization of grafted myofascia in the oral cavity was investigated using the latissimus dorsi muscle of Wistar strain rats. RESULTS: The use of myofascial tissue to fill in defects may maintain the space by preventing shrinkage and contracture of the wound, and this myofascial tissue may induce regeneration of the mucosal epithelium. Myofascia itself did not have the potential to epithelialize. Epithelialization progressed gradually along the surface of the granulated myofascia from the surrounding incised mucosal margin of the recipient site to the central portion. Immunohistochemical staining showed that keratinocyte growth factor-expressing cells were found mainly in the granulated myofascia. Within the study period, regenerated epithelium with orthokeratosis consisting of stratified squamous cells was thin and had scanty rete pegs, but it was similar to normal epithelium. CONCLUSION: Grafted myofascia in the oral cavity may play a space-making role, and induce regeneration of the mucosal epithelium, associated with the production of keratinocyte growth factor.

Animals↗
PubMed2003

Phylogeny, processing and expression of the rat cathelicidin rCRAMP: a model for innate antimicrobial peptides.

A database search identified a rat cDNA clone which phylogenetic analysis revealed to encode a cathelicidin most similar to mouse cathelicidin CRAMP. The analysis also showed that the evolutionary pattern of the cathelicidin family is lineage specific. The rat cathelicidin is called rCRAMP. Its peptide was isolated from granulocytes, and determined to be 43 amino acids long by mass spectrometry and N-terminal sequencing. Synthetic rCRAMP had antimicrobial activity. The expression of rCRAMP was investigated by reverse-transcriptase polymerase chain reaction followed by Southern hybridization and by Western blot analysis. rCRAMP was identified in granulocytes, thymus, testis, lung, mouth mucosa, tongue, oesophagus, colon, caecum and small intestine, a distribution similar to cathelicidins of mouse and human. The rat is a small laboratory animal with additional disease models available compared to the mouse. Our results open up the possibility to use the rat as a model system to study responses connected to cathelicidin expression in health and disease.

Amino Acid Sequence↗