Small interfering RNA targeting of keratin 17 reduces inflammation in imiquimod-induced psoriasis-like dermatitis / 中华医学杂志(英文版)

BACKGROUND@#Psoriasis is a common chronic inflammatory skin disease with 2% to 3% prevalence worldwide and a heavy social-psychological burden for patients and their families. As the exact pathogenesis of psoriasis is still unknown, the current treatment is far from satisfactory. Thus, there is an urgent need to find a more effective therapy for this disease. Keratin 17 (K17), a type I intermediate filament, is overexpressed in the psoriatic epidermis and plays a critical pathogenic role by stimulating T cells in psoriasis. Therefore, we hypothesized that inhibiting K17 may be a potential therapeutic approach for psoriasis. This study aimed to investigate the therapeutic effect of K17-specific small interfering RNA (siRNA) on mice with imiquimod (IMQ)-induced psoriasis-like dermatitis.@*METHODS@#Eight-week-old female BALB/c mice were administered a 5% IMQ cream on both ears to produce psoriatic dermatitis. On day 3, K17 siRNA was mixed with an emulsion matrix and applied topically to the left ears of the mice after IMQ application every day for 7 days. The right ears of the mice were treated in parallel with negative control (NC) siRNA. Inflammation was evaluated by gross ear thickness, histopathology, the infiltration of inflammatory cells (CD3+ T cells and neutrophils) using immunofluorescence, and the expression of cytokine production using real-time quantitative polymerase chain reaction. The obtained data were statistically evaluated by unpaired t-tests and a one-way analysis of variance.@*RESULTS@#The severity of IMQ-induced dermatitis on K17 siRNA-treated mice ears was significantly lower than that on NC siRNA-treated mice ears, as evidenced by the alleviated ear inflammation phenotype, including decreased ear thickness, infiltration of inflammatory cells (CD3+ T cells and neutrophils), and inflammatory cytokine/chemokine expression levels (interleukin 17 [IL-17], IL-22, IL-23, C-X-C motif chemokine ligand 1, and C-C motif chemokine ligand 20) (P < 0.05 vs. the Blank or NC siRNA groups). Compared to the NC siRNA treatment, the K17 siRNA treatment resulted in increased K1 and K10 expression, which are characteristic of keratinocyte differentiation (vs. NC siRNA, K17 siRNA1 group: K1, t = 4.782, P = 0.0050; K10, t = 3.365, P = 0.0120; K17 siRNA2 group: K1, t = 4.104, P = 0.0093; K10, t = 4.168, P = 0.0042; siRNA Mix group: K1, t = 3.065, P = 0.0221; K10, t = 10.83, P < 0.0001), and decreased K16 expression, which is characteristic of keratinocyte proliferation (vs. NC siRNA, K17 siRNA1 group: t = 4.156, P = 0.0043; K17 siRNA2 group: t = 2.834, P = 0.0253; siRNA Mix group: t = 2.734, P = 0.0250).@*CONCLUSIONS@#Inhibition of K17 expression by its specific siRNA significantly alleviated inflammation in mice with IMQ-induced psoriasis-like dermatitis. Thus, gene therapy targeting K17 may be a potential treatment approach for psoriasis.

Genetic study of a Chinese pedigree affected with pachyonychia congenita / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a Chinese pedigree affected with pachyonychia congenita (PC).@*METHODS@#With informed consent obtained, peripheral blood samples were taken from the pedigree. Genomic DNA was extracted with a phenol/chloroform method. Based on the clinical manifestation of the patients, candidate genes for PC were selected. Potential mutation was screened by PCR and Sanger sequencing. Suspected mutation was verified in other family members by PCR-high resolution melting (HRM) analysis. Haplotype analysis using microsatellite markers was also carried out to determine the founder of the mutation.@*RESULTS@#A heterozygous c.275A>G (Asn92Ser) mutation was discovered in exon 1 of the KRT17 gene in the proband. PCR-HRM analysis showed that all affected members were heterozygous carriers of the mutation. The same mutation was found in none of the unaffected members. Haplotype analysis and sequencing indicated the mother of the proband to be the founder.@*CONCLUSION@#The c.275A>G (Asn92Ser) mutation of the KRT17 gene probably underlies the disease in this pedigree. Above finding has facilitated genetic counseling and prenatal diagnosis for this pedigree.

Steatocystoma multiplex: A case report of a rare entity

Steatocystoma multiplex is an uncommon benign skin disease, which typically manifests as numerous intradermal cysts that can be scattered anywhere on the body. Although usually asymptomatic, it can be significantly disfiguring. One type of steatocystoma multiplex is known to be associated with the autosomal dominant inheritance of a mutation in the gene coding for keratin 17 (KRT17). In such cases, it is often concurrent with other developmental abnormalities of the ectoderm-derived tissues, such as the nails, hair, and teeth. To the best of our knowledge, few cases have been reported of steatocystoma multiplex of the oral and maxillofacial region. This report describes a case of steatocystoma multiplex of both sides of the neck and multiple dental anomalies, with a focus on its clinical, radiological, and histopathological characteristics, as well as the possibility that the patient exhibited the familial type of this condition.

Cytokeratin 13, Cytokeratin 17, and Ki-67 Expression in Human Acquired Cholesteatoma and Their Correlation With Its Destructive Capacity

OBJECTIVES: Cholesteatoma is a nonneoplastic destructive lesion of the temporal bone with debated pathogenesis and bone resorptive mechanism. Both molecular and cellular events chiefly master its activity. Continued research is necessary to clarify factors related to its aggressiveness. We aimed to investigate the expression of Ki-67, cytokeratin 13 (CK13) and cytokeratin 17 (CK17) in acquired nonrecurrent human cholesteatoma and correlate them with its bone destructive capacity. METHODS: A prospective quantitative immunohistochemical study was carried out using fresh acquired cholesteatoma tissues (n=19), collected during cholesteatoma surgery. Deep meatal skin tissues from the same patients were used as control (n=8). Cholesteatoma patients were divided into 2 groups and compared (invasive and noninvasive) according to a grading score for bone resorption based upon clinical, radiologic and intraoperative findings. To our knowledge, the role of CK17 in cholesteatoma aggressiveness was first investigated in this paper. RESULTS: Both Ki-67 and CK17 were significantly overexpressed in cholesteatoma than control tissues (P < 0.001 for both Ki-67 and CK17). In addition, Ki-67 and CK17 were significantly higher in the invasive group than noninvasive group of cholesteatoma (P=0.029, P=0.033, respectively). Furthermore, Ki-67 and CK17 showed a moderate positive correlation with bone erosion scores (r=0.547, P=0.015 and r=0.588, P=0.008, respectively). In terms of CK13, no significant difference was found between cholesteatoma and skin (P=0.766). CONCLUSION: Both Ki-67 and CK17 were overexpressed in cholesteatoma tissue and positively correlated with bone resorption activity. The concept that Ki-67 can be a predictor for aggressiveness of cholesteatoma was supported. In addition, this is the first study demonstrating CK17 as a favoring marker in the aggressiveness of acquired cholesteatoma.

Clinicopathologic features and immunohistochemistry of the basal-like subtype of invasive breast carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathologic features and immunohistochemical of the basal-like subtype of invasive breast carcinoma (BLBC), and to discuss the diagnosis standard.</p><p><b>METHODS</b>Immunohistochemistry was performed in 448 cases of breast carcinoma and these cases were categorized into luminal A, luminal B, null subtypes, HER2-overexpressing and basal-like and their clinicopathologic features were observed under light microscope with stains of HE and immunohistochemical InVitrogen staining.</p><p><b>RESULTS</b>Among the breast cancer patients, the incidence of BLBC was 15.4% (69/448). Morphologic features significantly associated with BLBC constituently included nest structure and showing diffuse growth pattern, large scarring areas without cells in tumor, geographic necrosis, pushing margin of invasion, lymphocytic infiltrate in various degree in tumor stroma, syncytial tumor cell without clear boundaries, tumor cell showing vesicular unclear chromatin and nucleolus, markedly elevated mitotic count, metaplasia (all P < 0.01). Meanwhile, most BLBC showed strong immunoreactivity for CK5/6, CK14, CK17 (all P < 0.01).</p><p><b>CONCLUSION</b>BLBC showed distinct morphologic and immunophenotypic features.</p>

A Novel Missense Mutation of Keratin 17 Gene in a Chinese Family with Steatocystoma Multiplex

No abstract available.

Establishment and characterization of a new carcinoma cell line from uterine cervix of Uyghur women / 中华病理学杂志

<p><b>OBJECTIVE</b>To establish a uterine cervical carcinoma cell line of Uyghur ethnical background and to evaluate the related biological characteristics for future biomedical investigations of diseases in the Uyghur population.</p><p><b>METHODS</b>Poorly-differentiated squamous cell carcinoma specimens of Uyghur patients were obtained and cultured in vitro by enzymatic digestion method, followed by continuous passaging to reach a stable growth determined by cell viability and growth curve. Morphological study, cell cycling and chromosomal analysis were performed. Tumorigenesis study was conducted by inoculation of nude mice. Biomarker (CK17, CD44, Ki-67, CK14 and vimentin) expression was detected by immunofluorescence and immunocytochemical techniques.</p><p><b>RESULTS</b>A cervical carcinoma cell line was successfully established and maintained for 12 months through 70 passages. The cell line had a stable growth with a population doubling time of 51.9 h. Flask method and double agar-agar assay showed that the cell line had colony-forming rates of 32.5% and 15.6%, respectively. Ultrastructural evaluation demonstrated numerous cell surface protrusions or microvilli, a large number of rod-shape structures in cytoplasm, typical desmosomes and nuclear atypia. Chromosomal analysis revealed human karyotype with the number of chromosomes per cell varying from 32 - 97 with a majority of 54 - 86 (60.3%). Xenogeneic tumors formed in nude mice showed histological structures identical to those of the primary tumor. The cells had high expression of CK17, CD44, Ki-67 and vimentin but no CK14 expression.</p><p><b>CONCLUSIONS</b>A cervical carcinoma cell line from a female Uyghur patient is successfully established. The cell line has the characteristics of human cervical squamous cell carcinoma, and it is stable with maintaining the characteristic biological and morphological features in vitro for more than 12 months, therefore, qualified as a stable cell line for further biomedical research.</p>

Expression of cytokeratin 17 in oral squamous cell carcinoma / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression of cytokeratin 17 (CK17) in oral squamous cell carcinoma (OSCC) as well as its clinical significance.</p><p><b>METHODS</b>Detection of the mRNA level and protein expression of CK17 in the in vitro cellular carcinogenesis model of OSCC, some OSCC cell lines and tissue specimens from 30 primary OSCC patients were performed using real-time polymerase chain reaction (PCR), Western blot and immunohistochemistry, respectively.</p><p><b>RESULTS</b>Increased CK17 mRNA level was observed in the HB56 and OSC cell lines compared with the HIOEC using real-time PCR technique. Western blot showed increased CK17 protein expression in all the cell lines compared with the HIOEC. Increased CK17 mRNA and immunoreaction levels were also observed in the cancerous tissue specimens from OSCC patients compared with normal adjacent tissues (P<0.01).</p><p><b>CONCLUSION</b>The significantly increased CK17 gene may be associated with the tumorigenesis and development of OSCC.</p>

Keratin 17 mutation in pachyonychia congenita type 2 in a Chinese Han family / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the keratin 17 gene (KRT17) mutation in a pedigree with pachyonychia congenita type 2 (PC-II).</p><p><b>METHODS</b>DNA was extracted from the blood samples of the patients, unaffected members of the pedigree, and 100 unrelated healthy controls. PCR was performed to amplify the hot spots in KRT17 gene. PCR products were directly sequenced to detect mutation.</p><p><b>RESULTS</b>A heterozygous 296T-->C mutation was found in all the affected members of this family, which resulted in the substitution of leucine by proline in codon 99 (L99P) in the 1A domain of the KRT17, but not in the healthy individuals from the family and the 100 unrelated controls.</p><p><b>CONCLUSION</b>The mutation of KRT17 may play a major role in the pathogenesis of this pedigree with pachyonychia congenita type 2.</p>

Expression of CK5/6 and CK17 and its correlation with prognosis of triple-negative breast cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the correlation of CK5/6 and CK17 expression with clinical outcome in patients with triple-negative [ER(-), PR(-), Her-2(-)] breast cancer.</p><p><b>METHODS</b>112 patients with breast cancer treated by surgery between 2000 and 2002 were included in this study. All cases were immunohistochemically proven to be triple-negative. Samples of formalin-fixed and paraffin-embedded surgical specimens were obtained for immunohistological examination for CK5/6 and CK17 expression. The correlation of the gene expression with clinicopathological features and outcome of the patients was analyzed.</p><p><b>RESULTS</b>Of the 112 triple-negative patients, five-year disease-free survival rate was 73.2% (82/112). The positive rate of both CK5/6 and CK17 was 21.4% (24/112), either CK5/6 or CK17 positive was 46.4% (52/112). It was shown by Kaplan-Meier curve that positive CK5/6, CK17 or CKs (CK5/6 or CK17 positive) was correlated with poor five-year disease-free survival (P = 0.020, P = 0.032, P = 0.003); and positive staining of CK5/6 or CKs was correlated with poor five-year overall survival (P = 0.027, P = 0.015). Of the 91 patients with invasive ductal carcinoma, a correlation of CK5/6 or CK17 positive staining with high grade differentiation was observed (P = 0.030), and with axillary lymph node metastasis was also noticed (P = 0.044). Multivariate analysis by Cox regression showed that differentiation grade, pathological stage and expression of CK5/6 were factors affecting both the disease-free-survival and overall-survival, while menopausal status was an independent factor affecting the disease-free-survival.</p><p><b>CONCLUSION</b>Positive expression of CK5/6 or CK17 in patients with triple-negative breast cancer is correlated with poor prognosis, high grade differentiation and axillary lymph node metastasis.</p>

Expressions of p53, Ki-67, PCNA and cytokeratin 17, cytokeratin 18 in recurred and non-recurred ameloblastoma

Steatocystoma Multiplex in a Family

Steatocystoma multiplex is a rare, inherited disorder that is characterized by multiple, asymptomatic, variably sized dermal cysts. The condition is transmitted in an autosomal dominant fashion; although sporadic cases have been documented. Keratin 17 has been proposed to be an important factor in inherited steatocystoma. In this study, a 29-year old man has a 4-year history of asymptomatic, movable, skin-colored nodules on his face, neck, scalp, anterior chest and back. His father and elder-brother have similar lesions. Histologically, the cysts show a thin stratified squamous epithelium with sebaceous glands arising from its wall and an absence of the granular cell layer. Generally, there are two treatments-medical treatment and surgical treatment. In case of non- inflamed lesions, surgical excision or drainage is regarded as the best treatment. We tried excisional biopsy and until now there has been no recurrence in the operation area over the past 12 months following the operation.

Clinical, Histopathological, and Immunohistochemical Study of Steatocystoma Multiplex / 대한피부과학회지

BACKGROUND: Steatocystoma multiplex is a rare but well-known disorder characterized clinically by the formation of numerous cutaneous cysts and histopathologically by a undulated hyalinized cuticle and sebaceous glands within or close to the cyst walls. However the clinical and histological features can be variable and may mimic other other cystic tumors. OBJECTIVE: The purpose of study was to characterize the clinical and histopathological features of steatocystoma multiplex and to assess the effectiveness of cytokeratin 10 and 17 stain in differentiating steatocystoma multiplex from epidermal cysts. METHODS: A total of 54 cases of SM diagnosed at the Department of Dermatology, Pusan National University Hospital(PNUH) were reviewed clinically and histopathologically. To compare the staining pattern of Cytokeratin 10 and 17 between steatocystoma multiplex and epidermal cyst, we used monoclonal mouse anti-human cytokeratin 10 and cytokeratin 17. RESULTS: 1. Male to female ratio is 1.4:1, and family history is identified in only 4 cases. 2. The mean age at visit and at onset of the lesions are 32 years(range:16 - 58) and 25.8 years(range: 13 - 58). 3. The major lesional sites at visit were the upper extremities(14 cases, 50%) and chest(13 cases, 46%), and those at onset were the upper extremities(7 cases, 25%) and axillae(6 cases, 22%). 4. Atypical histopathologic findings, such as round or oval cystic wall(25 cases, 46%), vellus hair in cystic wall(2 cases, 4%), foreign body granuloma reaction(1 case, 2%), or the cases not showing the diagnostic histopathologic findings(2 cases, 4%) are frequently noted. 5. Cytokeratin 10 and 17 immunohistochemical staining are helpful in discriminating steatocystoma multiplex from epidermal cyst. CONCLUSION: It is interesting that steatocystoma multiplex has more clinical and histopathological findings than the previous reports, and characteristic response to Cytokeratin 10 and 17.