Dysplasia Specimens Research Articles

Unsedated transnasal endoscopy accurately detects Barrett's metaplasia and dysplasia

Background: Unsedated transnasal upper endoscopy has a diagnostic yield comparable with that of sedated conventional upper endoscopy. The ability of transnasal upper endoscopy to detect Barrett's metaplasia or dysplastic change has not been systematically evaluated. The aim of this study was to assess the feasibility of transnasal upper endoscopy for surveillance of patients with Barrett's esophagus and to evaluate its histopathologic yield for Barrett's metaplasia and dysplasia. Methods: Thirty-two patients with Barrett's esophagus were evaluated with conventional upper endoscopy and transnasal upper endoscopy. An effort was made to recruit patients known to have dysplasia. Quadrantic biopsy specimens were taken with standard (conventional upper endoscopy) and pediatric (transnasal upper endoscopy) biopsy forceps at procedures performed at least 1 week apart. Two blinded pathologists evaluated the specimens. Results: Transnasal upper endoscopy detected Barrett's metaplasia histopathologically in 31 of 32 patients. Level of agreement for presence of dysplasia in biopsy specimens obtained between conventional upper endoscopy and transnasal upper endoscopy was excellent (k = 0.79). Interobserver agreement for dysplasia in specimens obtained by conventional upper endoscopy (k = 0.61) and by transnasal upper endoscopy (k = 0.61) were similar. Intraobserver agreement between conventional upper endoscopy and transnasal upper endoscopy biopsy specimens for dysplasia by pathologist 1 (k = 0.73) and pathologist 2 (k = 0.75) were also similar. No significant adverse effects were noted. Conclusions: Transnasal upper endoscopy is feasible and accurate for identification and histopathologic confirmation of Barrett's metaplasia with a histopathologic yield for dysplasia comparable with conventional upper endoscopy. (Gastrointest Endosc 2002;56:472-8.)

Subepithelial vascular patterns in bronchial dysplasias using a high magnification bronchovideoscope

Background: We have developed a method of high magnification bronchovideoscopy that enables improved observation of subepithelial vascular patterns of the bronchial mucosa. A study was undertaken to investigate the value...

Expression of inducible nitric oxide synthase in human oral premalignant epithelial lesions

Increased expression of inducible nitric oxide synthase (iNOS) has been found at the protein level in human oral epithelial dysplasias, but, a corresponding expression at the mRNA level has not been demonstrated. The purpose here was to assess the expression of iNOS mRNA and its correlation with the expression of the enzyme protein in human buccal premalignant epithelial lesions. Activities for iNOS protein (57/80, 64%) and mRNA (53/80, 53%) were detected in the specimens examined. Cytoplasmic and/or nuclear staining for iNOS protein was detected immunohistochemically in a number of mild oral epithelial dysplasias (16/20, 80%), moderate to severe oral epithelial dysplasias (14/20, 70%), submucous fibrosis (14/20, 70%) and verrucous hyperplasia (13/20, 65%). Upon in situ reverse transcription-polymerase chain reaction (RT-PCR), the cellular location of iNOS mRNA was compatible with the immunohistochemical findings. Furthermore, iNOS mRNA was found in 15 specimens of mild oral epithelial dysplasia (15/20, 75%), 13 specimens of moderate to severe oral epithelial dysplasia (13/20, 65%), 13 specimens of submucous fibrosis (13/20, 65%) and 12 specimens of verrucous hyperplasia (12/20, 60%). No iNOS protein or mRNA was found in samples of normal buccal mucosa, or in negative controls. Further studies on the characteristics of iNOS-positive cells and more long-term clinical follow-up data are needed.

Increased expression of the neuronal glutamate transporter (EAAT3/EAAC1) in hippocampal and neocortical epilepsy.

To define the changes in gene and protein expression of the neuronal glutamate transporter (EAAT3/EAAC1) in a rat model of temporal lobe epilepsy as well as in human hippocampal and neocortical epilepsy. The expression of EAAT3/EAAC1 mRNA was measured by reverse Northern blotting in single dissociated hippocampal dentate granule cells from rats with pilocarpine-induced temporal lobe epilepsy (TLE) and age-matched controls, in dentate granule cells from hippocampal surgical specimens from patients with TLE, and in dysplastic neurons microdissected from human focal cortical dysplasia specimens. Immunolabeling of rat and human hippocampi and cortical dysplasia tissue with EAAT3/EAAC1 antibodies served to corroborate the mRNA expression analysis. The expression of EAAT3/EAAC1 mRNA was increased by nearly threefold in dentate granule cells from rats with spontaneous seizures compared with dentate granule cells from control rats. EAAT3/EAAC1 mRNA levels also were high in human dentate granule cells from patients with TLE and were significantly elevated in dysplastic neurons in cortical dysplasia compared with non-dysplastic neurons from postmortem control tissue. No difference in expression of another glutamate transporter, EAAT2/GLT-1, was observed. Immunolabeling demonstrated that EAAT3/EAAC1 protein expression was enhanced in dentate granule cells from both rats and humans with TLE as well as in dysplastic neurons from human cortical dysplasia tissue. Elevations of EAAT3/EAAC1 mRNA and protein levels are present in neurons from hippocampus and neocortex in both rats and humans with epilepsy. Upregulation of EAAT3/EAAC1 in hippocampal and neocortical epilepsy may be an important modulator of extracellular glutamate concentrations and may occur as a response to recurrent seizures in these cell types.

High expression levels of nuclear factor κB, IκB kinase α and Akt kinase in squamous cell carcinoma of the oral cavity

BACKGROUND It has been reported that the transcription factor nuclear factor κB (NF-κB) is involved in the growth, invasion, and antiapoptotic activity of cultured tumor cells. METHODS The authors used immunohistochemistry to examine the expression of NF-κB and the signaling molecules leading to NF-κB activation in 36 untreated biopsy specimens from patients with squamous cell carcinoma (SCC) and in 15 specimens from patients with epithelial dysplasia of the oral cavity. RESULTS Among the molecules examined, the p65 subunit of NF-κB (p65) and IκB kinase α (IKKα) were expressed highly in almost all SCC specimens examined, whereas the samples of normal squamous epithelia adjacent to tumors as well as epithelial dysplasia specimens were negative in for immunohistochemical staining. The invasiveness and metastasis of SCC seemed to correlate with the degree of staining degree in the molecules. Moreover, phosphorylated Akt kinase, which may be associated with antiapoptosis signaling of NF-κB, was detected in the same areas where IKKα existed in large amounts. CONCLUSIONS The results suggest that high expression levels of p65 and IKKα contribute to malignant behavior and antiapoptotic activity in SCC of the oral squamous epithelium. Cancer 2001;92:3037–44. © 2001 American Cancer Society.

High expression levels of nuclear factor kappaB, IkappaB kinase alpha and Akt kinase in squamous cell carcinoma of the oral cavity.

It has been reported that the transcription factor nuclear factor kappaB (NF-kappaB) is involved in the growth, invasion, and antiapoptotic activity of cultured tumor cells. The authors used immunohistochemistry to examine the expression of NF-kappaB and the signaling molecules leading to NF-kappaB activation in 36 untreated biopsy specimens from patients with squamous cell carcinoma (SCC) and in 15 specimens from patients with epithelial dysplasia of the oral cavity. Among the molecules examined, the p65 subunit of NF-kappaB (p65) and IkappaB kinase alpha (IKKalpha) were expressed highly in almost all SCC specimens examined, whereas the samples of normal squamous epithelia adjacent to tumors as well as epithelial dysplasia specimens were negative in for immunohistochemical staining. The invasiveness and metastasis of SCC seemed to correlate with the degree of staining degree in the molecules. Moreover, phosphorylated Akt kinase, which may be associated with antiapoptosis signaling of NF-kappaB, was detected in the same areas where IKKalpha existed in large amounts. The results suggest that high expression levels of p65 and IKKalpha contribute to malignant behavior and antiapoptotic activity in SCC of the oral squamous epithelium.

Differential expression of glutamate and GABA-A receptor subunit mRNA in cortical dysplasia

Focal cortical dysplasia is characterized by disorganized cortical lamination, dysplastic and heterotopic neurons, and an association with epilepsy. The contribution that dysplastic and heterotopic neurons make to epileptogenesis in focal cortical dysplasia is unknown and the phenotype of these cells may be distinct. The authors hypothesized that the expression of genes encoding glutamatergic (glutamate [GluR] and N-methyl-D-aspartate NMDA receptors [NR]) and gamma-aminobutyric acid A receptor (GABA(A)R) subunits is distinct in dysplastic and heterotopic neurons and that changes in receptor gene expression could be defined in a cell-specific pattern. Single immunohistochemically labeled dysplastic and heterotopic neurons were microdissected from human focal cortical dysplasia specimens obtained during epilepsy surgery. Pyramidal neurons were microdissected from postmortem control cortex and from temporal cortex without dysplasia resected during temporal lobectomy. Poly (A) messenger RNA (mRNA) from single neurons was amplified, radiolabeled, and used to probe complementary DNA (cDNA) arrays containing GluR(1-6), NR(1A,1B), NR(2A-D), and GABA(A)Ralpha(1-6), and -Rbeta(1-3) subunit cDNAS: The relative hybridization intensities of each mRNA-cDNA hybrid were quantified by phosphorimaging. GluR, NR, and GABA(A)R subunit mRNA expression did not differ between control neurons and nondysplastic epilepsy specimens. Expression of GluR(4), NR(2B), and NR(2C) subunit mRNA was increased, and NR(2A) and GABA(A)Rbeta(1) subunit mRNA was decreased in dysplastic compared with pyramidal and heterotopic neurons. In contrast, GABA(A)Ralpha(1), -Ralpha(2), and -Rbeta(2) as well as GluR(1) mRNA levels were reduced in both dysplastic and heterotopic neurons. Differential expression of GluR, NR, and GABA(A)R mRNA in dysplastic and heterotopic neurons demonstrates cell specific gene transcription changes in focal cortical dysplasia. These results suggest that dysplastic and heterotopic neurons may be pharmacologically distinct and make differential contributions epileptogenesis in focal cortical dysplasia.

The role of cyclin-dependent kinase inhibitor p27 in squamous cell carcinoma of the esophagus.

The level of p27 expression decreases during tumor development and progression. Loss of p27 protein provides independent prognostic information in breast, prostate, colon, stomach and lung carcinomas. We generated a new polyclonal antibody against p27 and carried out immunohistochemical analysis of p27 expression in 61 specimens of esophageal squamous cell carcinoma, 10 carcinoma in situ specimens, and 5 squamous epithelial dysplasia specimens of the esophagus. We examined the correlation of p27 expression with various clinicopathologic features and prognosis. In squamous epithelial dysplasia, the p27-positive cells were located in the superficial normal-appearing cells, not in the atypical cells. In carcinoma in situ, the expression of p27 was markedly lower than in normal esophageal epithelium. In advanced squamous cell carcinomas, high p27 correlated significantly with the degree of differentiation (p = 0.0002) and the depth of tumor invasion (p = 0.001) was statistically significant. The high p27 expression group had a better prognosis than did the low or negative p27 expression groups, but these differences were not statistically significant. These observations may imply that a decrease in p27 expression occurs early on in the carcinogenesis of esophageal carcinoma. In addition, p27 expression may correlate with the histologic differentiation of squamous cell carcinoma of the esophagus.

High expression levels of nuclear factor ?B, I?B kinase ? and Akt kinase in squamous cell carcinoma of the oral cavity

It has been reported that the transcription factor nuclear factor kappaB (NF-kappaB) is involved in the growth, invasion, and antiapoptotic activity of cultured tumor cells.The authors used immunohistochemistry to examine the expression of NF-kappaB and the signaling molecules leading to NF-kappaB activation in 36 untreated biopsy specimens from patients with squamous cell carcinoma (SCC) and in 15 specimens from patients with epithelial dysplasia of the oral cavity.Among the molecules examined, the p65 subunit of NF-kappaB (p65) and IkappaB kinase alpha (IKKalpha) were expressed highly in almost all SCC specimens examined, whereas the samples of normal squamous epithelia adjacent to tumors as well as epithelial dysplasia specimens were negative in for immunohistochemical staining. The invasiveness and metastasis of SCC seemed to correlate with the degree of staining degree in the molecules. Moreover, phosphorylated Akt kinase, which may be associated with antiapoptosis signaling of NF-kappaB, was detected in the same areas where IKKalpha existed in large amounts.The results suggest that high expression levels of p65 and IKKalpha contribute to malignant behavior and antiapoptotic activity in SCC of the oral squamous epithelium.

High proliferative activity is associated with dysplasia in ulcerative colitis.

Ulcerative colitis is associated with an increased risk of colorectal neoplasia. Markers of proliferation are reported to be valuable in the diagnosis of dysplasia in ulcerative colitis. However, it is not known whether dysplastic change or proliferative change occurs first. Whether abnormal proliferation is present in normal-seeming mucosa in ulcerative colitis was investigated. Eighteen cancer or high-grade dysplasia specimens and 9 low-grade dysplasia specimens from 5 patients and 51 specimens from 31 patients without neoplasia were studied. Immunostaining with anti-Ki 67 antibody was used to evaluate proliferative activity. Labeling index (in the superficial one-half of crypt) was calculated. Crypts with labeling index more than 0.3 were determined to have abnormal proliferation. The mean +/- standard error of the mean labeling index in specimens negative for dysplasia (0.056+/-0.004) was significantly lower than that in low-grade dysplasia specimens (0.418+/-0.024) and that in high-grade dysplasia specimens (0.503+/-0.027; P < 0.0001). In specimens negative for dysplasia, only 4 (4 cases) of 339 (1.2 percent) crypts had abnormal proliferation, whereas the ratio of crypts with abnormal proliferation was 76 percent (54/71) in low-grade dysplasia and 92.1 percent (35/38) in high-grade dysplasia. The labeling index in background mucosa was 0.139+/-0.009, which was significantly higher than that in specimens negative for dysplasia (P < 0.001). In background mucosa 15.7 percent of crypts showed abnormal proliferation. A follow-up study revealed that two of four cases developed cancer or high-grade dysplasia one and seven years after proliferative abnormality was detected in nondysplastic specimens. Ki-67 immunostaining can be an aid in the diagnosis of dysplasia. High proliferating activity in background mucosa suggests that proliferating activity change precedes dysplasia detected with hematoxylin-and-eosin staining.

Telomerase activity in oral and maxillofacial tumors.

Telomerase activity was measured in biopsy specimens as well as surgically resected tissues of 39 oral squamous cell carcinomas (SCCs), 22 oral leukoplakias, 13 normal oral mucosas, 12 malignant salivary gland tumors, 10 benign salivary tumors and five normal salivary gland tissues adjacent to tumors using a polymerase chain reaction (PCR)-based telomerase assay. Telomerase activity was detectable in 38.5% (5/13) of normal oral mucosa samples, 54.5% (12/22) of leukoplakia samples, 82.1% (32/39) of oral SCC samples, 83.3% (10/12) of malignant salivary gland tumor samples and 0% (0/10, 0/5) of benign salivary gland tumor and normal salivary gland samples. High-level enzyme activities were seen in 20% (3/15) of mild dysplasia specimens, 50% (2/4) of moderate-severe dysplasia specimens, 48.7% (19/39) of oral SCC tissue specimens, and no high activity was seen in the normal mucosa and hyperkeratosis specimens (P for trend, <0.001). We also analyzed the proliferative activity of dysplastic leukoplakia, oral SCC, and salivary gland tumor specimens using Ki-67 immunohistochemistry. The Ki-67 labeling indices (LI) were significantly higher in dysplastic leukoplakia and oral SCC with high telomerase activity than in dysplastic leukoplakias and oral SCC with low and negative telomerase activity (P<0.01 and P<0.05). These results indicate that telomerase activity has some correlation with the progression of multistep oral carcinogenesis with the cellular proliferation, and also indicate that telomerase may be a specific marker used to distinguish malignant salivary gland tumors from their benign counterparts.

K-ras codon 12 mutations in Barrett's oesophagus and adenocarcinomas of the oesophagus and oesophagogastric junction.

Activation of the ras oncogene is commonly found in gastrointestinal tract cancers, but the role of ras in the development and progression of Barrett's oesophagus and associated cancers is uncertain. The frequency of K-ras codon 12 point mutations was assessed in 52 paraffin-embedded tissues from 44 patients with oesophageal pathology. The specimens were classified pathologically as follows: adenocarcinoma of the oesophagus or oesophagogastric junction (n = 23), Barrett's high-grade dysplasia (n = 5), low-grade dysplasia (n = 14), intestinal metaplasia (n = 4), normal oesophagus (n = 5) or normal stomach (n = 1). DNA was extracted from three consecutive sections of each paraffin block and mutations at bases 1 and 2 of K-ras codon 12 were identified using a novel restriction endonuclease-mediated selective polymerase chain reaction method. Mutations were found in 7 of 23 (30.4%) adenocarcinomas and in 2 of 5 (40%) high-grade dysplasia specimens. No mutations were found in specimens of low-grade dysplasia, intestinal metaplasia without dysplasia, or normal oesophagus and stomach. There were no significant associations between the presence of mutations and clinicopathologic features in the patients with cancer. One patient who progressed from low-grade to high-grade dysplasia was found to have developed mutant K-ras in the course of this transformation. These results suggest that K-ras codon 12 mutations may occur frequently in patients with Barrett's oesophagus with high-grade dysplasia or adenocarcinoma of the oesophagus or oesophagogastric junction. K-ras mutation may be a late event in the Barrett's metaplasia-dysplasia-adenocarcinoma sequence.

Reduced expression of cyclin-dependent kinase inhibitor p27Kipl is an indicator of malignant behavior in oral squamous cell carcinoma

Reduced expression of the cyclin-dependent kinase inhibitor P27Kip1 has been reported to correlate with poor survival in cohorts of breast and colorectal carcinoma patients. Post-translational ubiquitin-mediated proteasomal proteolysis is related to P27Kip1 protein levels. However, to the authors’ knowledge, no previous study has examined the expression of P27Kip1 in oral squamous cell carcinoma (OSCC). To examine the expression of P27Kip1 and its clinicopathologic roles in OSCC, the authors studied the expression of P27Kip1 protein by immunohistochemistry in deparaffinized tissue sections of 20 normal oral mucosa specimens, 22 epithelial dysplasia specimens and 70 OSCCs, and analyzed its correlation with clinicopathologic parameters. They also studied the expression of P27Kip1 mRNA and protein in six OSCC cell lines by Northern blot and Western blot analysis. To examine the mechanism of reduced expression of P27Kip1, OSCC cell lines were treated with the proteasome inhibitor LLnV. All the normal oral mucosa specimens and 73% (16 of 22) of the oral epithelial dysplasia specimens expressed P27Kip1 at high levels, whereas 87% of the OSCCs (61 of 70) showed reduced expression of P27Kip1. Furthermore, the levels of expression of this protein were significantly lower in carcinomas with metastasis than in those without metastasis. Although OSCC cell lines expressed P27Kip1 mRNA at various levels, most of them expressed P27Kip1 protein at lower or undetectable levels. LLnV induced the expression of P27Kip1 protein in HSC2 cells, in which P27Kip1 protein was originally undetectable. These findings suggest that 1) reduced expression of P27Kip1 may correlate with the development and progression of OSCC and can be an indicator of malignant behavior of this neoplasm, and 2) increased proteasome-mediated degradation may play an important role in the reduction of P27Kip1 protein expression. H. Tideman

Immunohistochemical study of MUC1 mucin in premalignant oral lesions and oral squamous cell carcinoma: association with disease progression, mode of invasion, and lymph node metastasis.

MUC1 mucin is a transmembrane, mucin-like glycoprotein encoded by the MUC1 gene. Although MUC1 expression has been identified in a variety of neoplastic tissues, to the authors' knowledge, few studies have examined MUC1 expression in premalignant and malignant oral lesions. A total of 36 specimens of oral epithelial dysplasia, 12 carcinoma in situ (CIS) specimens and 77 specimens of oral squamous cell carcinoma (OSCC), were examined by both light and electron microscopy using immunohistochemical staining of MUC1 mucin. Relations between staining patterns and clinicopathologic findings also were examined. Distinct membrane MUC1 mucin staining patterns were identified in epithelial dysplasia (33.0%), CIS (50.0%), and OSCC (59.7%) cases. A predominantly cytoplasmic staining pattern was detected in epithelial dysplasia (5.6%), CIS (41.7%), and OSCC (32.5%) cases. Significant positive correlations were found between MUC1 mucin membranous immunoreactivity and disease progression from epithelial dysplasia to OSCC (P < 0.01), mode of tumor invasion (P < 0.02), and lymph node metastasis (P < 0.01). Furthermore, the malignant transformation of oral epithelium, tumor invasion, and tumor metastasis were associated with higher MUC1 mucin expression in the cytoplasm (P < 0.01). In addition to the usual cell surface expression, cytoplasmic expression of MUC1 mucin was confirmed by colloidal gold labeling with transmission electron microscopy. The results of the current study suggest that determination of MUC1 mucin expression may be a parameter in the diagnosis of premalignant and malignant lesions arising in the oral cavity and that this expression may affect the malignant behavior of OSCC. MUC1 mucin expression may be a useful diagnostic marker for prediction of the invasive/metastatic potential of OSCC.

Histological assessment of apoptotic cell death in cardiomyopathies

Apoptosis in the myocardium is complex and often difficult to recognise. Myocyte apoptosis is scattered across the myocardial wall and is restricted to individual cells. In the present study, we describe the amount of apoptosis in 50 endomyocardial biopsies taken from 50 patients with dilated cardiomyopathy, in 14 hearts with hypertrophic cardiomyopathy and in five hearts with arrhythmogenic dysplasia of the right ventricle. As a control group, 15 endomyocardial biopsies from 15 transplanted hearts (of live patients) were used. Apoptosis was immunohistochemically determined in paraffin sections with the TUNEL method. In each specimen the TUNEL index was calculated as the percentage of TUNEL-positive nuclei among a total number of 200 counted nuclei. Cellular morphology was assessed in conjunction with TUNEL staining. The mean percentage of TUNEL-positive myocardial cells varied from 4% for dilated cardiomyopathy to 17.5% for arrhythmogenic right ventricle dysplasia and 18.5% for hypertrophic cardiomyopathy, whereas no signs of apoptotic myocardial cell death were found in normal subjects. The numbers of apoptotic cells in dilated cardiomyopathy specimens were significantly lower by comparison with both those of hypertrophic cardiomyopathy and those of arrhythmogenic right ventricular dysplasia specimens. It is evident that apoptosis constitutes a major biological phenomenon in the development of at least some heart diseases, but its role in their pathophysiology has yet to be delineated.

Grading dysplasia in colorectal adenomas by means of the quantitative binding pattern determination of Arachis hypogaea, dolichos biflorus, Amaranthus caudatus, Maackia amurensis, and Sambucus nigra agglutinins

The current study deals with the setting up of a new tool that enables the benign versus the malignant nature of colorectal adenomas to be determined accurately. The 2 objectives are to determine (1) whether adenomas should, or should not, be included in a 2- or a 3-tier grading system, and (2) whether severe dysplasias and carcinomas in situ share common or different biological characteristics. The levels of expression of different types of glycoconjugates were characterized in a series of 166 colorectal specimens, including 14 normal, 90 dysplastic, and 62 cancerous cases. The glycoconjugate expressions were demonstrated for 5 lectins, namely, Arachis hypogaea (PNA), Dolichos biflorus (DBA), Amaranthus caudatus (ACA), Maackia amurensis (MAA) and Sambucus nigra (SNA). The glycoconjugates demonstrated by these 5 lectins belong to the family of the Thomsen-Friedenreich antigens. The binding patterns of the 5 lectins were quantitatively determined by means of computer-assisted microscopy. The quantitative data were submitted to discriminant analyses. Our results show that the specific glycochemical staining patterns could be identified unambiguously and without misclassification between benign (normal and low dysplasia) and malignant (ie, either as moderate/severe dysplasia, carcinoma in situ, or cancer) cases. The data also strongly suggested that (1) dysplasias seem to be distinguishable in 2 instead of 3 groups, that is, low versus moderate/severe (high); and (2) moderate/severe dysplasias are biologically distinct from carcinomas in situ. The methodology developed can be applied directly in routine diagnosis to identify moderate/severe dysplasia specimens already exhibiting features common to carcinomas, and which therefore should be treated consistently in view of the fact that our data strongly suggest that most moderate/severe dysplasias are still benign, whereas carcinomas in situ are real carcinomatous lesions.

High-risk types of human papillomavirus associated with the progression of cervical dysplasia to carcinoma.

In the past several years, much evidence has accumulated that strongly implicates human papillomavirus (HPV) as an etiological agent of cervical cancer. However, the natural history of HPV infection is not yet completely understood, and at present there is no good marker to predict progression to invasive cancer in individual patients with dysplasia. Tissue specimens from 45 patients with cervical dysplasia were classified as showing progression to carcinoma (progression group; 26 women), spontaneous regression (regression group; 16 women), and persistence (persistence group; 3 women). The presence of HPV 16, 18, 33, and 52 DNA was examined in 90 formalin-fixed, paraffin-embedded surgical and biopsy specimens of dysplasia using the PCR method, and the relationship between the presence of HPV and cervical carcinogenesis was analyzed. The mean follow-up period was 25.8 months in the progressed group and 34.0 months in both the persistent and regressed groups. The mean ages were 46.4 in the progressed group and 46.8 in the persistent and regressed groups. HPV 16, 18, 33, and 52 DNA were detected in 19 of the 26 patients (73.1%) in the progressed group. All patients with HPV 16, 18, or 52 DNA showed progression to carcinoma. The HPV 33 DNA was detected in 5 of the 16 patients (31.3%) in the regressed group and in 2 of the 3 patients (66.7%) in the persistent group, while HPV 16, 18, and 52 DNA were not detected in the 19 patients that comprised the regressed and persistent groups. This retrospective study suggests that cervical dysplasia in patients with high-risk types of HPV possessed the potential to progress to carcinoma in situ and eventually to invasive carcinoma as well.

The association between epithelial proliferation and disease progression in the oral mucosa

The purpose of this study was to examine the possible association between epithelial proliferation and disease progression in the oral mucosa. Archival specimens of normal oral mucosa (n = 12), dysplasia (n = 17) and squamous cell carcinoma (n = 18) were sectioned and proliferating cells visualised by staining with Ki-67 antibody. The proliferative index of the epithelium (PI) was determined by total cell counts and point counting. Similar results were obtained using either method. Comparison of the three groups of tissues by one-way analysis of variance showed a significant increase in PI with increasing lesion severity (p < 0.001). The PI of both dysplasia and carcinoma groups was significantly higher than that of normal oral mucosa (p < 0.001). However, the difference between dysplasia and carcinoma groups was not significant. PI was not associated with tobacco or alcohol consumption. We therefore conclude that Ki-67 expression is an early marker of disease progression in the oral mucosa but, on its own, is not a good indicator of neoplastic transformation.

DNA ploidy of liver biopsies from patients with liver cirrhosis and hepatocellular carcinoma: a flow cytometric analysis

Flow cytometric DNA analysis was used to assess cellular kinetics of needle liver biopsies from patients with liver cirrhosis and hepatocellular carcinoma (HCC). An abnormal DNA content was shown in 44.5% of liver cirrhosis cases and in 78.6% of tumor sites. The number of proliferating cells (S+G2M%) was significantly increased in cirrhotic liver ( P<0.05). Dysplasia was found in 66% of cirrhotic specimens. All negative dysplasia specimens showed a diploid pattern while 69% of positive dysplastic specimens were aneuploid ( P<0.001). In conclusion, cell proliferation, aneuploidy and liver cell dysplasia are important indicators in liver cirrhosis for the development of HCC.

Reduced expression of cyclin-dependent kinase inhibitor p27Kip1 is an indicator of malignant behavior in oral squamous cell carcinoma.

Reduced expression of the cyclin-dependent kinase inhibitor p27Kip1 has been reported to correlate with poor survival in cohorts of breast and colorectal carcinoma patients. Posttranslational ubiquitin-mediated proteasomal proteolysis is related to p27Kip1 protein levels. However, to the authors' knowledge, no previous study has examined the expression of p27Kip1 in oral squamous cell carcinoma (OSCC). To examine the expression of p27Kip1 and its clinicopathologic roles in OSCC, the authors studied the expression of p27Kip1 protein by immunohistochemistry in deparaffinized tissue sections of 20 normal oral mucosa specimens, 22 epithelial dysplasia specimens, and 70 OSCCs, and analyzed its correlation with clinicopathologic parameters. They also studied the expression of p27Kip1 mRNA and protein in six OSCC cell lines by Northern blot and Western blot analysis. To examine the mechanism of reduced expression of p27Kip1, OSCC cell lines were treated with the proteasome inhibitor LLnV. All the normal oral mucosa specimens and 73% (16 of 22) of the oral epithelial dysplasia specimens expressed p27Kip1 at high levels, whereas 87% of the OSCCs (61 of 70) showed reduced expression of p27Kip1. Furthermore, the levels of expression of this protein were significantly lower in carcinomas with metastasis than those without metastasis. Although OSCC cell lines expressed p27Kip1 mRNA at various levels, most of them expressed p27Kip1 protein at lower or undetectable levels. LLnV induced the expression of p27Kip1 protein in HSC2 cells, in which p27Kip1 protein was originally undetectable. These findings suggest that 1) reduced expression of p27Kip1 may correlate with the development and progression of OSCC and can be an indicator of malignant behavior of this neoplasm, and 2) increased proteasome-mediated degradation may play an important role in the reduction of p27Kip1 protein expression.