Expression of cytokeratins (CKs) 8, 13 and 18 and their mRNA in epithelial linings of radicular cysts: implication for the same CK profiles as nasal columnar epithelium in squamous epithelial lining.

OBJECTIVE: Our objective was to specify cytokeratin (CK) patterns in lining epithelia of radicular cysts which are sometime lined with ciliated columnar epithelia as seen in the nasal epithelia. MATERIALS AND METHODS: We examined the CK expression in 52 radicular cysts obtained from 32 maxillary and 20 mandibular lesions and investigated CK-mRNA expression using in situ hybridization in 24 maxillary and 13 mandibular cysts and reverse transcription-polymerase chain reaction (RT-PCR) in 24 maxillary cysts. RESULTS: Of the maxillary cysts, 20, 29 and 19 squamous epithelial linings were positive for CK8, CK13 and CK18, respectively; of the mandibular cysts, 10, 20 and 11 linings were positive for these CKs, respectively. The expression patterns of CK18(+)-CK13(-), CK18(+)-CK13(+) and CK18(-)-CK13(+) were observed in 3, 16 and 13 linings of the maxillary cysts and 0, 11 and 9 linings of the mandibular cysts, respectively. In situ hybridization revealed the expression of CK18-mRNA in 9 and 4 linings of 24 maxillary and 13 mandibular cysts examined, respectively. With RT-PCR, we explored that both CK18- and CK13-mRNA were expressed not only in the normal nasal and gingival epithelia but also in the examined maxillary cyst linings although their expression levels differed correlating with the difference in CK staining. CONCLUSION: It is concluded that CK13- and CK18-mRNA are constitutively expressed in columnar and squamous epithelial cells, respectively, and that the variant CK expression patterns with CK18-mRNA expression in maxillary radicular cysts are indicative of the possibility of phenotypic transformation in the cyst linings.

Mn-SOD antisense upregulates in vivo apoptosis of squamous cell carcinoma cells by anticancer drugs and gamma-rays regulating expression of the BCL-2 family proteins, COX-2 and p21.

ROIs and their scavengers are associated with apoptosis induction by anticancer drugs and gamma-rays, but the details have not been clarified. We examined the effect of transfection of Mn-SOD antisense on apoptosis by 5-FU, PLM, CDDP and gamma-rays using nu/nu mice. After inoculation of Mn-SOD antisense-transfected SCC cells into the subcutis of each mouse's back, they slowly multiplied to form tumors sized 1,460 +/- 70 mm(3) at day 60, while control vector-transfected SCC cells rapidly multiplied, with a mean tumor size of 2,330 +/- 220 mm(3). Inversely, mice in the Mn-SOD antisense group survived longer (mean survival duration 94.4 +/- 12.7 days) compared to those in the empty vector group (67.3 +/- 6.8 days). After treatment with 5-FU (5 microg/day), PLM (50 microg/day), CDDP (10 microg/day) and gamma-rays (2 Gy/day), mean survival times were largely prolonged, to 126.3 +/- 22.7, 123.0 +/- 22.1, 136.3 +/- 24.0 and 143.0 +/- 20.8 days, respectively, while mean survival times in the empty vector group were 91.7 +/- 14.8, 85.7 +/- 13.3, 97.5 +/- 16.0 and 100.7 +/- 17.1 days, respectively. Immunohistologically, tumors in the Mn-SOD antisense group revealed additional nick end-labeled cells compared to those in the empty vector group. In comparison, strong expression of Bax, Bak and p21(waf1/cip1) and suppressed expression of Bcl-2, Bcl-X(L) and COX-2 were observed in the Mn-SOD antisense group and the expression pattern of these proteins was the inverse in the empty vector group. The increased expression of these proapoptotic proteins appeared to be p53-independent because p53 protein expression was not increased in the antisense group. These immunohistologic results were supported by Western blotting of each protein. In conclusion, Mn-SOD antisense transfection is advantageous for apoptosis induction of SCC cells by anticancer drugs and gamma-rays through induction of proapoptotic Bcl-2 family proteins and suppression of antiapoptotic protein expression.

Peplomycin, a bleomycin derivative, induces myofibroblasts in pulmonary fibrosis.

To analyse the mechanism by which a bleomycin derivative, peplomycin (PLM) induces pulmonary fibrosis, we investigated differentiation of rat pulmonary fibroblasts to myofibroblasts (MF). In intraperitoneally PLM (5 mg/kg/day)-injected rats, the peripheries of lungs adjacent to the pleura revealed advanced fibrosis with a small number of alpha-smooth muscle actin (alpha-SMA)-positive MF, which ultrastructurally possessed abundant microfilaments and cellular organelles. In the fibrotic tissue, the expression of alpha-SMA-mRNA was detected by in situ reverse transcription-polymerase (RT-PCR). The message was strong just after a 2-week administration of PLM then decreased thereafter, although fibrosis advanced. When pulmonary fibroblasts were separated from saline-injected rats (N-Fib) and cultivated for 7 days in the presence of 5 mg/mL PLM, alpha-SMA protein was weakly expressed, while the majority of pulmonary fibroblasts separated from PLM-injected rats (P-Fib) became positive for alpha-SMA in 7-day cultivation and the expression of alpha-SMA in P-Fib was strongly increased by cultivation in the presence of PLM and transforming growth factor-beta (TGF-beta), but not basic fibroblast growth factor (bFGF) or platelet-derived growth factor (PDGF), although the cell proliferation was most strongly enhanced by bFGF and only slightly by PLM and TGF-beta. The alpha-SMA-positive cells expressed vimentin, but only weakly expressed desmin. Additionally, P-Fib generated larger amounts of TGF-beta and bFGF than were generated by N-Fib. These results indicate that PLM induces pulmonary fibrosis by differentiating fibroblasts to alpha-SMA-positive MF, and that bFGF and TGF-beta play each critical role in the different phases of PLM-induced pulmonary fibrosis by inducing fibroblast proliferation and transformation, respectively.

Diffuse mode of tumor cell invasion and expression of mutant p53 protein but not of p21 protein are correlated with treatment failure in oral carcinomas and their metastatic foci.

Lack of control of metastatic foci is the most prevalent cause of death in patients with oral carcinomas, and it is important for tumor control to identify the factors that predispose patients to death. In the present study, we examined 225 patients with oral squamous cell carcinoma and investigated the immunohistopathological characteristics of 43 tumors that led to death, comparing them with those of the non-lethal tumors. In the 43 patients, lack of control of the primary site, lymph node and distant metastatic tumors were noted in 20, 18 and 16 patients, respectively. The mode of tumor cell invasion was closely correlated with death. The diffuse invasion modes of grades 4C and 4D were observed in 15 (34.9%) of the 43 tumors with a poor outcome and in 35 (19.2%) of the 182 controlled tumors (p < 0.02). The expression of p53 was highly correlated with death. Of the tumors with poor prognosis, p53 protein was expressed in 32 tumors (76.2%). However, p53 protein expression was observed in 52.7% of the tumors with good prognosis (p < 0.02). In contrast, the expression of p21 protein in the well-controlled tumors (30.4%) was almost equal to that of the 43 lethal tumors (26.2%). Compared with the ratios of local recurrence, metastases and their treatment failures in the p53-negative grade 1 and 2 tumors, those in the mutant p53-positive grade 3, 4C and 4D tumors were mostly high. These results indicate that measuring p53 protein expression and evaluating the mode of tumor cell invasion are important for oral carcinoma therapy because the expression of mutant p53 protein and the diffuse modes of tumor cell invasion indicate a predisposition toward a poor prognosis.

Risk factors of metastasis in oral squamous cell carcinomas.

Lymph node and distant metastasis were comparatively studied in 225 oral carcinomas, and factors predisposing toward metastasis were investigated using clinical and immunohistopathological approaches. Neither the sites of tumors nor T-stage was correlated with either type of metastasis. Tumor cell differentiation was weakly correlated with lymph node metastasis, and stromal reaction (the degree of cell infiltration) did not differ greatly between metastasis-positive and negative tumors, although natural killer (NK) activities were correlated with lymph node metastasis. However, the mode of tumor cell invasion was closely associated with both lymph node and distant metastases. In grade 4C and 4D tumors, distant and lymph node metastases were observed in 8 (16%) and 31 (62%) cases, respectively, while of 68 grade 1 and 2 tumors, distant metastasis was not observed in any, and lymph node metastasis occurred in only 15 (22. 1%). In addition, the expression of p53 protein was correlated with lymph node metastasis; of 70 tumors without p53 protein expression, 23 (32.9%) revealed lymph node metastasis, while it occurred in 54 out of 96 tumors positive for p53 protein. However, p53 protein expression was not associated with distant metastasis, and p24 protein, a cyclin-dependent kinase inhibitor, did not show any relationship with either type of metastasis. These results indicate that lymph node metastasis is correlated with multiple factors in the host and tumor cells, but distant metastasis is only correlated with the mode of tumor cell invasion, suggesting that the former can be highly accurately predicted by invasion mode, p53 protein expression and NK activity.

Expression of p53 and p21 proteins in oral squamous cell carcinoma: correlation with lymph node metastasis and response to chemoradiotherapy.

p53 protein, a product of the p53 cancer suppressor gene, and p21 protein, a cyclin-dependent kinase inhibitor, were immunohistochemically investigated in 150 oral squamous cell carcinomas (SCCs) and the relationship between their expression and clinicopathological findings were evaluated. The positivity for p53 and p21 proteins was not correlated with the T-stage, mode of tumor cell invasion or tumor cell differentiation. However, the expression of p53 and p21 proteins was correlated with lymph node metastasis. Of 62 SCCs with regional lymph node metastasis, 45 SCCs (72.6%) were positive for p53 while 45 (52.9%) of 88 SCCs without metastasis expressed p53 protein (p < 0.02). In addition, p21 protein was observed in 25 (38.5%) and 18 (21.2%) SCCs with and without metastasis, respectively (p < 0.05). Furthermore, p53 protein was inversely correlated with the histopathological effect of inductive chemoradiotherapy; the rate of chemoradiotherapy-induced lethal degeneration (56.7%) in p53-negative SCCs was significantly higher than that (28.9%) in p53-positive SCCs (p < 0.005). However, no clear difference in the effect was observed between p21-positive and p21-negative SCCs. Finally, the 5-year-survival rate was highest in p53(-)-p21(+) (80.0%) followed by 76.3% in p53(-)-p21(-), 65.9% in p53(+)-p21(+) and 65.4% in p53(+)-21p(-) SCCs. These results indicate that although the expression of p21 protein is only weakly correlated with the clinico-histopathological findings, p53 protein is a useful prognostic marker and that inductive chemoradiotherapy can be successfully planned by immunohistochemical examination of p53 protein.

Hemostatic treatment after tooth extraction in a patient with factor V deficiency.

BACKGROUND: Factor V deficiency is a very rare hereditary coagulation disorder and tooth extraction in the patient with factor V deficiency has not been reported except in one case. PATIENTS AND TREATMENT: A 38-year-old woman with factor V deficiency was referred for extraction of the impacted lower third molar. After intravenous administration of frozen fresh plasma (FFP) and recognition of an increase of factor V level from 1-31%, upper and lower third molars were extracted. Eighteen and 48 h after the extraction, factor V was intermittently supplemented by injection of 4 and 2 units of FFP, respectively, and factor V was maintained above 12%. To form fast coagula and to protect the wound, the lower extraction socket was filled with a fibrin glue composed of factor XIII and fibrinogen (Beriplast P) and a plastic splint was applied. The wound was healed and epithelized within 2 weeks after the extraction without any bleeding or infectious consequences. CONCLUSION: Extraction in the patient with factor V hereditary deficiency is safely performed by both supplementation of factor V and application of local hemostasis.

Low malignant intraductal carcinoma on the hard palate: a variant of salivary duct carcinoma?

A rare, minor salivary gland tumour of the hard palate in a middle-aged woman was presented. The small (1.0 X 0.5 cm in diameter) hemispherical tumour was well circumscribed with a fine papillomatous surface. Histopathologically, tumour cells with eosinophilic cytoplasm and a large nucleus were single-strand cuboidal and columnar cells, which showed intraductal growth exhibiting a cribriform pattern. The histological features were distinct from adenoid cystic carcinoma and polymorphous low-grade adenocarcinoma because the tumour lacked the neurotropic infiltration, cord-like proliferation and targetoid arrangement. The tumour could not be identified as a typical salivary-duct carcinoma because Roman bridging, papillary projection, and severe cell atypia were not found. Tumour cells were negative for PAS, Alcian blue, mucicarmine, p53, c-erbB-2, CEA, S-100 protein, alpha-smooth muscle actin, lactoferrin or vimentin. About 5% of the tumour cells were positive for proliferating cell nuclear antigen. Taking these factors into account, together with the clinical features, the name low malignant intraductal carcinoma seems appropriate.

Benign and malignant myoepithelioma of salivary-gland - an immunohistochemical evaluation.

Three myoepitheliomas (MEOs) derived from the salivary glands were examined immunohistochemically. Proliferating cell nuclear antigen (PCNA)-positive cells were very rare (less than 2% of all tumor cells) in localized tumors of case 1 (epithelioid) (E-oid) cells) and case 2 (plasmacytoid) (P-toid) cells with a small number of spindle-shaped cells), but the percentage of PCNA-positive cells was high (21.8%) in case 3 (clear cells) exhibiting bone destruction. Strong c-myc expression was detected in all the tumors, but p53 or c-erbB-2 protein was not detected in any of the cases. More than half of the clear cells were positive for epidermal growth factor (EGF), while fewer tumor cells in cases 1 and 2 expressed EGF. A few tumor cells in cases 2 and 3 were positive for EGF-receptor (R). Keratin was most prominent in the E-oid cells, The P-toid cells were most strongly positive for S-100 protein followed by the E-oid and clear cells. More than half of the spindle-shaped cells and one-third of the E-oid cells were positive for alpha-smooth muscle actin (alpha-SMA), but less than 5% of the clear cells and none of the P-toid cells were positive for alpha-SMA. These results suggest that tumor cells in MEO are heterogenous and have different proliferation activities.

Tumorigenicity of cell lines established from oral squamous cell carcinoma and its metastatic lymph nodes.

We investigated the biological and histopathological characteristics of seven human tumour cell lines established from primary tongue squamous cell carcinoma (OSC-1), from metastasised lymph nodes of the gingival carcinoma (OSC-2 and OSC-3) and from tongue carcinoma (remaining four lines). The doubling time ranged from 22 h (OSC-2 and OSC-4) to 55 h (OSC-7), and did not correlate with tumour cell stratification in a collagen gel matrix. An invasive tendency was most prominent in OSC-2 and OSC-4; with the other cell lines, except OSC-6 and OSC-7, only a few sporadic invading cells were found in the tissue culture. In the cell lines established from the metastatised tumours, originally exhibiting grade 3 invasion, the invasion became more sporadic when the tumour cells were xenografted into the tongues of nude mice, while an invasion similar to the original was observed in the cell lines obtained from the original site (OSC-1) and from tumours of Grade 4C invasion. These findings suggest that the biological behaviour of the established tumour cells is markedly different even in tumours of the same tissue origin, and strongly invasive cells may selectively invade, and metastatise to the lymph nodes.

Lectin immunohistochemical evaluation of human bladder carcinomas. A comparison of Carnoy's and formalin fixation.

A lectin immunohistochemical analysis of 51 human bladder carcinomas, including 44 cases of transitional cell carcinoma (TCC) (G1, 15 cases; G2, 17 cases; G3, 12 cases) and 7 cases of squamous cell carcinoma (SCC), was performed. Tissues were obtained by cold punch biopsies, fixed in Carnoy's or 10% formalin solution, stained for binding of 10 different lectins, and evaluated under the light microscope. The lectins used were concanavalin agglutinin (Con A), soybean agglutinin (SBA), Lotus tetragonolobus agglutinin (LTA), Dolichos biflorusa agglutinin (DBA), peanut agglutinin (PNA), Ricinus communis agglutinin I (RCA1), Ulex europaeus agglutinin I, II (UEA-I, II), wheat germ agglutinin (WGA), and Pisum sativum agglutinin (PEA). TCC prepared with Carnoy's fixation tended to show moderately positive Con A, UEA-I, and WGA reactions for G1, and strongly positive reactions for G2 and G3 lesions. UEA-II was mainly negative in G1, but tended to increase to become moderate in G3. DBA tended to show a moderately positive reaction in G1 and G2, but was mainly negative in G3. With formalin fixation, only RCA1 demonstrated grade specific variation, tendency to react moderately in the G1 and G2 cases, and strongly in G3. There were no further differences among the histopathological grades of TCC for other lectins. Thus, Carnoy's fixation appears superior for distinguishing between grades of lesions. SCC tended to react more strongly than TCC with all the various lectins except PEA, independent of fixation.

Immunohistochemical evaluation of mam-3 and mam-6 antigens in squamous-cell carcinoma of the oral cavity.

MAM-3 and MAM-6 antigens of human milk fat globule membrane designated as human polymorphic epithelial mucin (PEM) were detected immunohistochemically in a total of 86 specimens from 39 cases of oral squamous cell carcinoma (SCC) and 6 cases of normal oral mucosa. The two antigens were identified by MoAbs 67D11 and 115G3 (MAM-3 antigen) and by MoAbs 115D8 and 115F5 (MAM-6 antigen) in paraffin sections. MoAb 115G3 staining was found in well keratinized foci; whereas, MoAb 115D8 and 115F5 staining was very low in keratinized tumor cells. The cytochemical distribution of the antigens was classified into 3 types; (i) cell membrane positive type, (ii) whole cell positive type and (iii) antigen negative type. Reduction and disappearance of the two antigens were correlated to the mode of tumor invasion and suggested that glycocalyx complex in cell membrane properties was changed for malignant transformation.

Erythropoietin expressed in granular convoluted tubule cells of mice submandibular glands under hypoxia, anemia, and nephrectomy.

Immunohistochemically detectable erythropoietin-like substance(Epo) in granular convoluted tubule(GCT) cells of submandibular glands (SMG's) was examined in mice in which hemolytic anemia had been induced by phenylhydrazine (ph), and in mice subjected to hypoxia, nephrectomy, or testosterone (TP) injections. Staining for Epo was negative in GCT cells of SMG's in normal mice, while positive staining occurred in GCT cells of the anemic mice and mice subjected to hypoxia or nephrectomy. A positive Epo reaction was also revealed at the luminal borders of distal tubules, and in cells of proximal and distal tubules in the kidney, and in some hepatic and spleen cells, of mice that had received combination regimens producing anemia and hypoxia, or had been nephrectomized. Increased staining of Epo was found in GCT cells of SMG's, and in proximal and distal kidney tubules of mice given the combination treatment plus TP injections. The detection of Epo in GCT cells suggests these extrarenal cells to be sites of accumulation or biosynthesis of the protein under certain specific conditions such as hemolytic anemia and hypoxia.

Mucinous adenocarcinoma of the submandibular gland.

A rare tumor not easily classifiable among published histologic categories for salivary gland tumors is reported. The neoplasm developed within the submandibular gland of a 78-year-old woman with invasion of the mandible and metastasis to regional lymph nodes. Histopathologically, cuboidal cells possessing clear cytoplasm and displaced round nuclei proliferated and exhibited an adenomatous pattern. Many cystic spaces surrounded by tumor cell strands were seen, mucus substance filled in the cystic spaces, and the tumor cells seemed mucus-secreting, but neither epidermoid cells nor papillary appearance could be observed. Electromicroscopically, numerous mucous droplets of low electron density were prominent in the cytoplasm, and the tumor cells had sparse irregular microvilli on the luminal surface. Mucin histochemistry, including paradoxical concanavalin A staining, revealed that the tumor cells contained neutral and acid mucins, and these were identified as class II and III mucosubstances. No other neoplastic lesion, except recurrent metastatic neck nodes, has been detected 6 years after the first examination, and it seems that the tumor is a rare primary mucinous adenocarcinoma of the submandibular gland.

Immunohistochemical staining of infiltrates in oral lichen planus.

Immunohistochemical examination of lymphocyte and accessory cell infiltrates was performed in 24 cases of oral lichen planus using a double staining technique. The major population of the superficial stromal lymphocytes was T cells which were mostly composed of dominant CD 4+Leu8- (helper T cells) and lesser numbers of CD 8+11b- cells (cytotoxic T cells). Contrarily, many more CD 8+11b- cells than CD 4+Leu8- cells had infiltrated the epithelium. Some infiltrated T cells expressed interleukin-2 receptor and about half of cytotoxic T cells expressed class II antigen. HLA-DR-positive monocytes were also observed in both the superficial stroma and the epithelium. A number of HLA-DR-bearing CD 1+ cells (Langerhans/dendritic cells) and CD 11c+ cells (macrophages) were observed in the lower layers of the epithelium which were sometimes degenerative. Having significant correlation with the infiltration intensities of subepithelial macrophages and epithelial CD 8+ cells (cytotoxic/suppressor T cells), epithelial Langerhans cells variably infiltrated. Superficial stromal CD 8+ cells correlated with epidermal CD 8+ and CD 4+ cell (helper/inducer T cells) infiltrates. These findings are consistent with the notion that Langerhans cells and macrophages play an important role in antigen presentation, and suggest that cellular immunity, mediated by cytotoxic T cells with helper T cells, may be related to the pathogenesis of oral lichen planus.

Langerhans cells in odontogenic tumours and cysts as detected by S-100 protein immunohistochemistry.

Immunohistochemical demonstration of S-100 protein in Langerhans cells (LCs) was made in odontogenic epithelial tumours (71 cases), radicular cysts (40 cases), follicular cysts (28 cases), odontogenic keratocysts (11 cases), primordial cysts (7 cases) and fissual cysts (6 cases). With the use of polyclonal antiserum against S-100 protein, positive LCs, dendrical or irregular in shape were found in tumour or cystic epithelia, and sometimes in stromal connective tissue. Incidence of positive S-100 staining LCs was 11 cases out of 61 ameloblastomas, 22 cases out of 40 radicular cysts, 3 cases of 28 follicular cysts, and other lesions in both odontogenic tumours and cystic diseases lacked LCs. The cases with S-100 protein positive LCs were usually accompanied with a high degree of inflammatory infiltration in their lesions; on the contrary, the negative cases also generally lacked inflammatory responses.

Histochemical studies on irradiated and obstructive human submandibular glands. Protein distribution and lectin-binding in the degenerative glands.

Immunohistochemical protein distribution of alpha-amylase (Am), lysozyme (Ly), cytokeratinin (CK), S-100 protein (S-100) and secretory component (SC), and lectin-binding (SBA and UEA-I) profiles were studied in 10 obstructive and 20 irradiated human submandibular glands which were surgically extirpated. Degenerative intensity of the glands was graded as I, II and III based on the order of severity. All proteins generally existed in serous acinic cells of the intact glands. The proteins immunoreactivities became weak even in mildly inflamed glands (grade I), and nearly disappeared from the moderately damaged glands (grade II). Duct cells had clear CK and some cells reacted with the anti-SC antibody, but other proteins were not observed on the ducts. Mucous cells possessed none of the proteins, and their lectin-binding was only traceable in some glands. Compared with immunoreactivities in the proteins, lectin-binding profiles were different. SBA and UEA-I bound somewhat similarly to both acinic and duct cells, and the binding was hardly affected even by severe degeneration (grade III). Between obstructive and irradiated glands, no obvious difference was observed in either protein distribution or lectin-binding. From the above, it seems that some proteins are more affective to the degeneration and that lectin-binding sugar residues are non-affective against the degenerative changes of the tissues.

Immunostaining of involucrin in odontogenic epithelial tumors and cysts.

An immunoperoxidase method was used to detect involucrin in 47 odontogenic tumors and 35 radicular cysts. Of a total of 40 ameloblastomas, 9 cases were positive for involucrin expression and those positive cases exhibited acanthomatous or follicular patterns. Squamous odontogenic tumors were strongly positive for involucrin, whereas adenomatoid odontogenic tumors gave a negative staining reaction. Involucrin expression in odontogenic tumors was divided into three categories: single cell positive, focally positive, and squamous metaplastic cell positive. Radicular cysts showed a very irregular distribution of involucrin; nonstratified epithelium was generally negative or showed only trace staining for involucrin, whereas suprabasilar stratified squamous epithelial cells were strongly positive. Cells positive for involucrin in odontogenic tumors and in cystic epithelium are probably direct signs of epithelial differentiation; such cells were squamoid in appearance.

Immunohistochemical evaluation of nonspecific cross reactive antigen and carcinoembryonic antigen (CEA) in urinary bladder carcinoma.

The immunohistochemical expression of CEA in formalin-fixed paraffin sections in urinary bladder carcinomas was compared to the use of polyclonal anti-CEA antiserum (P-CEA), NCA-absorbed anti-CEA (NCA-aCEA) and monoclonal antibody to CEA (M-CEA). The urinary bladder carcinomas examined consisted of 19 cases of transitional cell carcinoma (TCC) and 7 cases of squamous cell carcinoma (SCC). Both TCC and SCC were positive for CEA with the use of P-CEA and NCA-aCEA, and the degree of staining was markedly dependent on the grade of malignancy in TCC. However, the reaction to M-CEA was generally very weak or negative in TCC and SCC. In SCC, the staining reaction was confined to keratinized foci and not found in all malignant tumour cells when polyclonal CEA antiserum was used. These findings indicate that positive reactions seen with conventional CEA antibodies (P-CEA and NCA-aCEA) are possibly related to NCA and that urinary bladder carcinoma may contain relatively more NCA than true CEA.

Immunohistochemical expression of keratin proteins in urinary bladder carcinoma.

Transitional carcinomas of the urinary bladder were examined immunohistochemically for keratin proteins with the use of polyclonal antiserum (TK, 41-65 kDa) and 3 monoclonal antibodies (KL 1, 55-57 kDa; PKK 1, nos. 19, 18, 8; and K 8.12, nos. 16, 13). Umbrella cells gave particularly strong staining for TK, KL 1 and PKK 1, whereas they were negative for K 8.12. Basal- and intermediate-layer cells in urothelial epithelium were moderately positive for all keratins. Brunn's nests cells showed comparatively slight or moderate keratin staining, and K 8.12 staining of Brunn's nests was higher than in urothelial epithelial cells. Transitional carcinoma (grades I and II) indicated uniform keratin distribution, and staining was strong with TK, while that of KL 1, PKK 1 and K 8.12 varied, and grade III tumors showed the lowest intensity of staining. K 8.12 staining in papillary transitional carcinomas was strongly positive in basal located tumor cells, as compared with apical tumor cells. Squamous cell carcinoma was varying positive to keratin reactions dependent on the degree of keratinization. Heterogenity of keratin distribution in papillary transitional carcinomas was given between basal tumor cells and well differentiated tumor cells including umbrella-like cells.