involucrin and Carcinoma-in-Situ

Hematopoietic pre-B-cell leukemia transcription factor-interacting protein (HPIP) is a corepressor of pre-B-cell leukemia homeobox (PBX) 1 and is known to play a role in hematopoiesis. Recently, HPIP was demonstrated to promote breast cancer cell proliferation and hepatocellular carcinoma growth. Moreover, it has been revealed that homeobox and PBX proteins, the expression of which is regulated by HPIP, play key roles in cancer of various organs, including oral squamous cell carcinoma (OSCC). Nevertheless, there has not been any study regarding the role of HPIP in OSCC. This study investigated the expression of HPIP in normal oral mucosa, epithelial precursor lesion (OEPL), and OSCC, and the functional roles of HPIP in OSCC cells and normal keratinocytes.. Immunohistochemical analysis of HPIP, Ki-67, and involucrin was performed in OSCC specimens, and the change in involucrin expression following RNA interference treatment against HPIP was examined by quantitative RT-PCR and Western blot analysis in SCC9 and NHEK cells undergoing extracellular calcium-induced differentiation. Matrigel transwell and cell proliferation assays for both cell lines transfected with HPIP siRNA were also conducted.. HPIP expression increased in OEPL and OSCC specimens. In vitro analysis revealed that HPIP suppressed differentiation and proliferation of SCC9 cells and transwell migration of NHEK cells, while HPIP promoted invasion of SCC9 and proliferation of NHEK cells. However, HPIP has no significant effect on NHEK cell differentiation.. HPIP may play a critical role in oral carcinogenesis and is thus a potential target for anticancer therapy, with particular emphasis on its involvement in differentiation and migration/metastasis.

Topics: Adult; Aged; Calcium; Carcinogenesis; Carcinoma in Situ; Carcinoma, Squamous Cell; Cell Culture Techniques; Cell Differentiation; Cell Line, Tumor; Cell Movement; Cell Proliferation; Co-Repressor Proteins; Female; Gene Silencing; Humans; Keratinocytes; Ki-67 Antigen; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Protein Precursors; RNA, Small Interfering; Transcription Factors

Transcription factor c-Jun plays a key role in controlling epithelium cell proliferation, apoptosis and differentiation. However, molecular mechanism and biological functions of c-Jun in squamous differentiation and the progression of esophageal squamous cell carcinoma (ESCC) remain elusive. In this study, we found that c-Jun bound directly to the promoter region, and activated the transcription of differentiation-associated genes including cystatin A, involucrin and SPRR3 in vivo. Ectopic expression of c-Jun enhanced SPRR3 transactivation in KYSE450 cells. Conversely, TAM67, a dominant negative mutant of c-Jun, inhibited SPRR3 transactivation. c-Jun increased expression of SPPR3 mainly via a PKC/JNK pathway in response to TPA in KYSE450 cells. Furthermore, c-Jun was remarkably reduced in esophageal cancer. Interestingly, cystatin A, involucrin and SPRR3 were significantly downregulated as well, and associated with differentiation grade. Expression of c-Jun was correlated with the expression of these genes in normal epithelium and ESCC. Importantly, the expression of these genes was remarkably decreased during the malignant transformation from normal epithelium to low-grade intraepithelial neoplasia (LGIN) or high-grade intraepithelial neoplasia (HGIN). The expression of cystatin A and involucrin was significantly reduced from LGIN to HGIN. These results suggest c-Jun was involved in the regulation of differentiation-associated genes in ESCC. These genes might serve as the potential markers in distinguishing normal epithelium from esophageal squamous intraepithelial neoplasia.

Topics: Aged; Carcinoma in Situ; Carcinoma, Squamous Cell; Cell Differentiation; Cell Line; Cornified Envelope Proline-Rich Proteins; Cystatin A; Disease Progression; Epithelium; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Female; Gene Expression Regulation, Neoplastic; Genes, Dominant; Genes, jun; Humans; Male; Middle Aged; Mutation; Oligonucleotide Array Sequence Analysis; Protein Kinase C; Protein Precursors; Proto-Oncogene Proteins c-jun; Transcriptional Activation

Keratinization of the ocular surface epithelium is associated with various disorders impairing vision. We immunohistochemically determined whether the ocular surface epithelia express involucrin, and whether its expression pattern may differ in benign vs. malignant disorders. Expression of cytokeratins was also examined to provide further information relative to the epithelial differentiation.. We evaluated 17 specimens; 6 specimens of the normal ocular surface epithelia, 3 specimens from cases of conjunctival intraepithelial neoplasia (CIN), 6 of conjunctival squamous cell carcinoma (SCC) and 2 of conjunctivae from cases of superior limbic keratoconjunctivitis (SLK).. Corneal epithelium exhibited intracellular immunoreactivity for involucrin. Four of the 6 specimens of bulbar conjunctival epithelium showed involucrin immunoreactivity in the perimembranous region, whereas the fornical conjunctiva was negative. Cornified envelope in SLK specimens was positive for involucrin. The CIN showed its immunoreactivity in the perimembranous region in all levels of the hyperproliferative epithelium without keratinization, i.e., similar to the bulbar conjunctiva. The neoplastic cells of well-differentiated SCC showed involucrin in the perimembranous region, and those of moderately- to poorly-differentiated SCC have involucrin in their cytoplasm. The expression pattern of cytokeratins was unrelated to grade of malignancy in ocular SCC.. The epithelia of normal subjects and of CIN expresses involucrin without keratinization. In contrary, the keratinized SLK epithelium markedly expresses involucrin in the cornified envelope. The subcellular immunolocalization of involucrin in the ocular SCC may help in evaluating the differentiation, i.e., malignancy, of neoplastic cells.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma in Situ; Carcinoma, Squamous Cell; Conjunctiva; Conjunctival Diseases; Conjunctival Neoplasms; Epithelium; Eye Proteins; Female; Filaggrin Proteins; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratins; Keratoconjunctivitis; Male; Middle Aged; Protein Precursors

The immunoperoxidase method for involucrin detection was applied to the study of the maturation of epithelial lesions of the oral mucosa that included specimens of leukoplakia, lichen planus, verrucous carcinoma, carcinoma in situ and invasive carcinoma. Areas of orthokeratinized, parakeratinized, and non-keratinized normal mucosa were also studied. Normal orthokeratinized epithelia showed intracytoplasmic or pericellular staining in the suprabasal epithelial layers in a pattern similar to that of the normal epidermis. Parakeratinized and non-keratinized epithelia were less stained. Intense staining was observed in leukoplakia, whereas the staining of lichen planus was less intense but exhibited a more homogeneous pericellular staining pattern than leukoplakia. Verrucous carcinoma was markedly and very irregularly stained. Carcinomas in situ and invasive carcinoma exhibited a slightly positive and patchy reaction. The distribution patterns of involucrin in the lesions correlated very well with the degree of epithelial differentiation. In addition, irregular patchy distribution correlated with the degree of atypia, and was especially evident in carcinomas.

Topics: Carcinoma; Carcinoma in Situ; Carcinoma, Papillary; Humans; Immunoenzyme Techniques; Leukoplakia, Oral; Lichen Planus; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms; Protein Precursors

Involucrin is a precursor of the cross-linked envelope protein of the human stratum corneum. Its appearance in the upper layers of the epidermis reflects normal differentiation of keratinocytes. This study uses an immunoperoxidase technique for localization of involucrin in paraffin sections of normal conjunctiva, conjunctival dysplasia, carcinoma in situ, and invasive carcinoma. Clinicoimmunocytochemical correlations are presented. The results demonstrate that the distribution patterns of involucrin differ in precancerous and cancerous conjunctival lesions: normal limbal conjunctiva shows involucrin only in the three superficial cell layers; the fornix conjunctivae contains no involucrin. All 23 conjunctival dysplasias show an involvement also of deeper layers of the epithelium, sparing the basal layers. Three carcinomas in situ and one invasive squamous cell carcinoma, however, demonstrate an involvement of all layers of the epithelium. The involucrin staining pattern helps in histologic differential diagnosis of epithelial lesions of the conjunctiva.

Topics: Biopsy; Carcinoma; Carcinoma in Situ; Carcinoma, Squamous Cell; Conjunctiva; Conjunctival Neoplasms; Epithelium; Humans; Immunoenzyme Techniques; Neoplasm Invasiveness; Precancerous Conditions; Protein Precursors

Ninety-three cervical conization specimens with condyloma or intraepithelial neoplasia were stained by the peroxidase-antiperoxidase technique for involucrin. Diffuse, homogeneous suprabasal staining was observed in the ectocervical squamous mucosa and mature squamous metaplasia. In immature squamous metaplasia, staining was limited to cells with apparent squamous differentiation. Although diffusely reactive in the upper layers of condyloma and cervical intraepithelial neoplasia (CIN) grade I, the stain was uneven in the former and lacking in the parabasal layers of the latter. The staining intensity, distribution, and pattern were more variable in CIN grade II and grade III. With increasing severity, a patchy pattern with a mixture of reactive and nonreactive cells predominated. Although immunoreactivity with involucrin could not distinguish immature squamous metaplasia from neoplasia, the staining patterns in CIN correlated with extent of disease, degree of squamous differentiation, and cellular disorganization.

Topics: Carcinoma in Situ; Carcinoma, Squamous Cell; Condylomata Acuminata; Female; Humans; Immunoenzyme Techniques; Protein Precursors; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Involucrin is a soluble protein precursor of the cross-linked envelope present in the submembranous zone of human stratum corneum, and subsequently demonstrated in stratified squamous epithelia. The immunoperoxidase technique was used to assess the distribution of involucrin in 107 normal and 318 abnormal tissues. With few exceptions, involucrin was restricted to squamous epithelia, urothelium, some skin appendages and thymic Hassall's corpuscles. In normal squamous epithelium and normal urothelium, staining was most intense in the superficial layers where it was concentrated at the cell periphery and gradually decreased toward the basal layer. This orderly staining pattern was maintained in benign squamous and urothelial lesions and in grade I papillary urothelial carcinomas. Higher grade papillary urothelial carcinomas, infiltrating urothelial and squamous carcinomas, and in situ urothelial and squamous carcinomas demonstrated abnormal staining patterns for involucrin that are described. Foci of squamous differentiation in adenocarcinomas and other epithelial malignancies stained intensely for involucrin. Brenner tumours of the ovary and Walthard rests of the fallopian tube, lesions of uncertain histogenesis but possibly urothelial-related, also stained for involucrin. Results of this study suggest that involucrin is a sensitive and specific marker for squamous and urothelial differentiation, staining patterns for involucrin may be helpful in distinguishing benign from malignant urothelial and squamous lesions, and presence of involucrin may be helpful in determining the histogenesis of selected lesions.

Topics: Carcinoma in Situ; Carcinoma, Squamous Cell; Cell Differentiation; Cell Transformation, Neoplastic; Epithelium; Humans; Immunoenzyme Techniques; Neoplasms; Protein Precursors; Urinary Bladder Neoplasms

Forty-two cervical biopsies with cervical intraepithelial neoplasia were compared with respect to the expression of human papillomavirus (HPV) structural proteins and the expression of the cellular structural protein involucrin, a marker of suprabasal squamous differentiation. HPV structural protein and involucrin expression displayed an inverse correlation with the severity of dysplasia. Both of these proteins were detected in 11 of 28 cases (39%) of mild and moderate dysplasia, but in only two of 14 (14%) cases of severe dysplasia. This difference was statistically significant (p less than 0.001). The presence of HPV was also associated with expression of involucrin in the full thickness of the epithelium, including the basal layer, and an altered staining pattern in the more superficial cells, particularly the koilocytotic cells. These findings support the hypothesis that squamous differentiation is required for the expression of viral structural proteins and that HPV infection begins in the basal epithelium. The study also demonstrates the utility of involucrin staining in differentiating virus-induced cytologic atypia from true neoplasia.

Topics: Adult; Animals; Carcinoma in Situ; Cell Differentiation; Female; Humans; Papillomaviridae; Protein Precursors; Tumor Virus Infections; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms; Viral Proteins; Viral Structural Proteins