Odontogenic Keratocysts Don't Harbor BRAF Mutation: A Genetic and Immunohistochemical Analysis.

Odontogenic tumors (OTs) are important lesions of the gnathic bones due to their clinicopathological heterogeneity and variable biological behavior; therefore, epidemiological studies are needed to outline the incidence and behavior of these tumors. To evaluate the incidence and epidemiological profile of ameloblastoma (AMB) and keratocystic odontogenic tumor (KCOT) from an oral pathology service, and correlate morphological findings of these tumors with the immunoexpression of a cellular proliferation marker (Ki-67), a retrospective study (2002-2012) was conducted to characterize demographic, clinical, radiological, and morphological data of AMBs and KCOTs. Then, a representative sample composed of 49 cases of each tumor was selected to perform immunohistochemical (IHC) analysis of Ki-67 through the streptavidin biotin peroxidase technique. For statistical analysis, we used Fisher's exact test (p<0.05). A total of 279 OTs were found in the service, in which 91 (32.6%) were AMB and 98 (35%) were KCOT. Most cases occurred in white women, and the average age of patients with AMB and KCOT was 32 and 33 years, respectively. The maxilla-mandible ratio was 1:6 and 1:3.6 for AMB and KCOT, respectively. Regarding IHC analysis, AMB and KCOT had similar levels of cellular proliferation. However, KCOTs with intense inflammation showed higher Ki-67 expression (p<0.001). Recurrent cases had similar Ki-67 immunoexpression. The demographic profile of the studied tumors corroborates with data reported in the literature, and the levels of cellular proliferation were similar in both tumors, although the inflammation seems to induce a differential proliferative behavior in KCOT.

A keratocystic odontogenic tumor (KCOT) is a benign destructive recurrent odontogenic cystic neoplasm. The microRNAs (miRNAs) miR-15a and miR-16-1 function as negative regulators of the anti-apoptotic gene BCL2 at the post-transcriptional level. Notably, high Bcl-2 immunoexpression is found in the epithelial lining of KCOTs, while the loss of Bcl-2 immunopositive cells is observed in marsupialized cysts. The purpose of this study was to investigate whether the transcription of miR-15a and miR-16-1 is altered in KCOTs and whether it is associated with BCL2 gene expression in such lesions. Using qRT-PCR and immunohistochemical analyses, we examined miR-15a/16-1 and BCL2 gene expression in KCOTs. The impact of miR-15a/16-1 expression on BCL2 gene translation was investigated by in vitro studies using primary KCOT culture cells. Using qRT-PCR, we observed miR-15a and/or miR-16-1 downregulation in the majority of the KCOT samples (24 of 28). We also observed higher BCL2 mRNA expression in 19 of 20 KCOT frozen samples and moderate to high Bcl-2 immunopositivity in the basal layer cells of 16 of 18 paraffin embedded KCOTs (median: 42.6 %). In vitro over-expression of miR-15a/16-1 in human KCOT-1 primary cell cultures resulted in a decrease in Bcl-2 protein expression. Furthermore, all five paired KCOTs collected before and after marsupialization treatment exhibited an increase in miR-15a after the procedure. Our results suggest that KCOT neoplastic cells exhibit an anti-apoptotic profile that may be related to lower miR-15a/16-1 expression. Additionally, we demonstrated that miRNA expression increases after marsupialization, implicating an etiological and therapeutic role of miRNAs in KCOT.

Increased expression of autophagy-related proteins in keratocystic odontogenic tumours: its possible association with growth potential

Evaluation and comparison of odontogenic keratocysts and detigerous cysts immunoexpression and immunostaining intensities of Ki-67 antigen by assessing the whole extent of the epithelium (all epithelium layers in combination) and each layer individually. Ki-67 immunoexpression was evaluated in 15 odontogenic keratocysts and 6 dentigerous cysts using automated methods and the Aperio Technologies Inc. computer system. No statistically significant differences were observed in immunoexpression nor in immunostaining intensities between both lesions. Also, no statistically significant differences were found between odontogenic keratocysts from maxilla versus mandible nor primary versus recurrent. However, odontogenic keratocyst showed a significantly higher cellular proliferation index in the suprabasal layers compared to the basal layer. Assessment of the cellular proliferation index through a computerized system enabled the evaluation of all epithelial tissue without field selection. The increased Ki-67 immunoexpression in suprabasal layers of odontogenic keratocyst suggests a different biological behavior and more aggressive proliferation potential when compared to dentigerous cyst. The same result was found in recurrent odontogenic keratocysts when compared with primary ones. The odontogenic keratocysts of the maxilla and mandible have similar Ki-67 immunoexpression. The evaluation of cellular proliferation only by immunohistochemical analysis with Ki-67 antigen does not provide enough data to elucidate the biological behavior of odontogenic keratocyst.

Odontogenic keratocyst (OKC) first described by Philipsen in 1956 constitutes approximately 11% of all cysts of the jaws. Adenomatoid odontogenic tumour (AOT) is an uncommon, benign epithelial lesion of odontogenic origin. The aim of this study was to analyse the expression of Bcl-2 in OKC and its comparison with other selected benign odontogenic tumours (OTs). Ten formalin fixed paraffin embedded blocks of OKCs, five each of AOT and unicystic ameloblastoma Bcl-2 protein is characterized by its ability to inhibit apoptosis. OKC were characterized by higher expression of Bcl-2 in basal cell epithelium. AOT and unicystic ameloblastoma differed from OKC in a wide spectrum of apoptosis and/or cell cycle-related protein expressions, higher proliferation in the basal cell layer, and vice versa, lower proliferation in the suprabasal cell layer. The solitary OKC seems to be less biologically aggressive and should be classified as a cyst rather than a tumour, means that at least few of OKCs manifests as ordinary cysts. Some of the present study findings could support the theory that OKCs are with high proliferative, probably that these lesions are developmental cysts with some neoplastic properties because of the high intrinsic growth potential. WHO recommends the term KCOT as it better reflects the neoplastic nature of the lesion; however, this reclassification has not yet been universally accepted.

The odontogenic keratocyst (OKC) is a controversial lesion that was reclassified as a tumor with the name “keratocystic odontogenic tumor” in 2005. The reclassification was revoked recently in 2017, with a conclusion on the need for further studies on the subject. In this study, the expressions of an important regulatory protein (maspin), an important integral membrane proteoglycan (syndecan-1), and a universal proliferation marker (Ki-67) in the epithelium of the OKC were investigated in comparison with the dentigerous cyst (DC) and ameloblastoma (AB). Twenty-six OKCs, eleven DCs, and ten conventional ABs were immunohistochemically stained for maspin, syndecan-1, and Ki-67. ImageJ was used to analyze the positivity of maspin and syndecan-1. The Ki-67 score was calculated as the percentage of positive nuclei in 5 high power fields. Analysis of variance (ANOVA) test and Student t-test were used as appropriate. Lower expressions of maspin were noted in OKC and DC compared to those in AB, and lower expressions of syndecan-1 were noted in OKC and AB compared to those in DC. The differences, however, did not reach statistical significance (ANOVA and t-test: P > 0.05). The Ki-67 score was significantly higher in OKC than in DC (t-test: P < 0.05), and not significantly different from AB (t-test: P > 0.05). In conclusion, expressions of maspin and syndecan-1 are not strongly representative of differences in behavior between OKC, AB, and DC. However, the expression of Ki-67 indicates comparable proliferative activities of OKC and AB, which are higher than that of DC. Further investigation on the biologic behavior of OKC is still recommended to arrive at more specific conclusions regarding its classification.

Background: Odontogenic cysts are characterized by their sluggish growth and ability to enlarge, primarily affecting the oral and maxillofacial tissues. Timely diagnosis and treatment are crucial to prevent potentially serious consequences. The present study aimed to evaluate and compare the immunohistochemical expression of cytokeratin 19 in the epithelium of odontogenic keratocyst, dentigerous, and radicular cysts. Methods: This study analyzed forty-five formalin-fixed, paraffin-embedded tissue blocks containing odontogenic cysts. The sample consisted of fifteen odontogenic keratocysts, fifteen dentigerous cysts, and fifteen radicular cysts. Immunohistochemical analysis was performed to assess the expression of the cytokeratin 19 epithelial marker in these samples. Statistical analyses were conducted using Statistical Package for Social Sciences version 26, employing the Chi-square test for comparative analysis of cytokeratin 19 expression among the odontogenic keratocyst, dentigerous cyst, and radicular cyst. Results: This study showed that 80% of basal layer tissue samples in the odontogenic keratocyst group had negative cytokeratin 19 biomarker scores. In contrast, 60% of dentigerous cyst tissue blocks and 53.3% of radicular cyst tissue blocks were +1. This difference was statistically significant (P = 0.034). In comparison, between groups, there was no significant difference (P = 0.103) in CK19 expression in the surface and spinous layers. Conclusion: The level of epithelial differentiation is correlated with cytokeratin 19 expression. The cysts with well-differentiated epithelium (radicular cyst and dentigerous cyst) express cytokeratin 19, whereas those with less well-differentiated epithelium (odontogenic keratocyst) exhibit low positives. Thus, it serves as a diagnostic tool for distinguishing these three lesions.

Frontiers Events is a rapidly growing calendar management system dedicated to the scheduling of academic events. This includes announcements and invitations, participant listings and search functionality, abstract handling and publication, related events and post-event exchanges. Whether an organizer or participant, make your event a Frontiers Event!

The aim of this study was to evaluate the biological profile of odontogenic epithelium by immunolabeling of epidermal growth factor receptor (EGFR), Ki-67 and survivin in keratocystic odontogenic tumors (KOT), dentigerous cysts (DC), and pericoronal follicles (PF). Immunohistochemical analysis was performed in 13 KOTs, 14 DCs and 9 PFs. Immunolabeling was analyzed in the basal and suprabasal layers of KOTs and DCs, and in the islands of odontogenic epithelium and/or reduced enamel epithelium of PFs. KOTs showed the highest proliferation rate among the three groups, mainly in suprabasal layers. EGFR immunolabeling was observed mainly in the cytoplasm in basal and suprabasal layers of KOTs and in the suprabasal layer of DCs. Immunolabeling in both membrane and cytoplasm was greater in PFs. In PFs, membrane-only staining was observed. Survivin immunolabeling showed a greater percentage of positive cells (scoring +++) in the suprabasal layer of KOTs. In DCs, both layers showed similar percentages of cells scoring +++; PFs showed the highest percentage of these cells. In KOTs, epithelial cells showed stimulus-independent neoplastic proliferative characteristics, suggesting the presence of a suprabasal proliferative compartment, maintained by inhibition of apoptosis. In DCs, the basal layer seemed to proliferate in response to stimulus. Although PFs showed low proliferative activity, the expression of EGFR indicates that some cells have a high capacity to respond to stimuli, which could probably explain the origin of odontogenic lesions.

The aim of the current study was to investigate the presence of human papillomavirus (HPV) and evaluate its association with Ki-67 and cyclooxygenase-2 (COX-2) expressions in keratocystic odontogenic tumor (KCOT). Nineteen cases were included in the present study. Conventional PCR method and immunohistochemical analysis were performed for the detection of HPV-DNA and HPV-L1 capsid protein. Moreover, the expressions of Ki-67 and COX-2 proteins were analyzed immunohistochemically. HPV-DNA was detected in 36.8% (7/19) of tumor samples, whilst HPV-L1 protein was identified in 68.4% (13/19) of them. The Kappa coefficient statistical test showed a moderate agreement (κ 0.424) between PCR and IHC assays for HPV detection. Expression of HPV-DNA was positively correlated with Ki-67 and COX-2 expressions (p < 0.05), whereas HPV-L1 positive staining was positively correlated with COX-2 (p < 0.05) and highly associated with those of Ki-67 (p < 0.01). There was no significant correlation between the presence of HPV and the recurrence of the studied lesions. The results of the current study showed that active HPV infection was present in the odontogenic epithelium of KCOT, and it was associated with increased proliferation rate and COX-2 expression. These findings suggest that HPV may have a role in the pathogenesis and aggressiveness of KCOT. Based on these conclusions, we recommend further investigations of HPV vaccine or antiviral therapy and COX-2 inhibitors as nonsurgical options in the prevention and management of KCOT.

We examined the immunohistochemical expressions of cell-cycle- and apoptosis-related factors to investigate the possible role of these factors in odontogenic keratocyst (OKC). Expression of cyclin D1 and p16 protein was detected in the basal and parabasal cells in lining epithelium of OKCs and was found more frequently in basal cell nevus syndrome (BCNS)-associated OKCs than in primary or recurrent OKCs. Positivity for p21 protein was detected in basal to superficial cells, whereas that for p27 protein was detected in parabasal to superficial cells in lining epithelium of OKCs. DNA topoisomerase IIalpha reacted with nuclei in basal and parabasal cells of the lining epithelium of OKCs, and positive cells were observed in BCNS-associated OKCs significantly more frequently than in primary or recurrent OKCs. Expression of Fas in suprabasal to superficial cells and expression of Fas-L in basal and parabasal cells were detected in lining epithelium of OKCs. Immunoreactivity for caspase-3 was detected in basal to suprabasal or superficial cells in lining epithelium of OKCs. Single stranded (ss)DNA-positive nuclei were detected in superficial cells in lining epithelium of OKCs. Fas was more broadly distributed in BCNS-associated OKCs than in primary OKCs, and ssDNA-positive cells were observed in BCNS-associated OKCs significantly more frequently than in primary or recurrent OKCs. These results suggest that BCNS-associated OKCs might be a distinguishable entity from solitary OKCs.

Odontogenic keratocysts constitute 10%-20% of odontogenic cysts and exhibit a distinctive corrugated parakeratinized lining epithelium. Considering that cornified envelope formation is an important phenomenon during keratinocyte differentiation, this study aimed to clarify the characteristics of cornified envelope formation in odontogenic keratocysts. We investigated the cellular distribution of cornified envelope-related proteins (transglutaminases and their substrates), as well as the upstream regulatory protein c-Fos, by immunohistochemical analysis of the lining epithelium of 20 odontogenic keratocysts. We examined the corresponding mRNA levels by quantitative polymerase chain reaction. Ten dentigerous cysts served as control non-keratinized cysts. The distributions of transglutaminase and their substrates except loricrin and small protein-rich protein 1a significantly differed between odontogenic keratocysts and dentigerous cysts. There was no significant difference in c-Fos expression between odontogenic keratocysts and dentigerous cysts. The mRNA levels of transglutaminases and their substrates were significantly higher in odontogenic keratocysts than in dentigerous cysts. However, c-Fos mRNA levels did not significantly differ between groups. Surprisingly, the overall appearance of cornified envelope-related proteins of odontogenic keratocysts was consistent with the characteristics of non-keratinized oral mucosa identified in previous studies. These findings indicate that the contribution of cornified envelope-related molecules in odontogenic keratocysts is similar to that in non-keratinized oral epithelium, rather than keratinized oral epithelium, suggesting that odontogenic keratocysts are not genuine keratinized cysts. The upregulation of cornified envelope-related genes in odontogenic epithelium could be an important pathognomonic event during odontogenic keratocyst development.

Background: Odontogenic keratocyst (OKC) is one of the common odontogenic cysts with aggressive clinical behavior and a high recurrence rate. Epithelial–mesenchymal transition (EMT) is a process, in which the epithelial cell loses its epithelial characteristics and acquires mesenchymal features. Since the evidence for the involvement of EMT in the development of OKC is still limited, the present study aimed to investigate the immunohistochemical expression of EMT-related proteins (E-cadherin and N-cadherin) in OKC and compare them to radicular cyst (RC) and dentigerous cyst (DC). Materials and Methods: In this descriptive analytical study, 75 paraffin blocks, including 25 DCs, 25 OKC, and 25 RCs, were selected. Immunohistochemical staining was performed to determine the expression and staining intensity of E-cadherin and N-cadherin proteins. The specimens were examined under an optical microscope, and the data were analyzed using the Kruskal–Wallis test in SPSS statistical software (version 23) with a significance level of 5%. Results: The expression of N-cadherin in OKC was higher than that in other cysts; nonetheless, there was no statistically significant difference (P = 0.331). The staining intensity of N-cadherin was weak in most cases, and this difference was not statistically significant (P = 0.252). E-cadherin expression in OKC was significantly lower than that in radicular and DCs (P = 0.003). In addition, the staining intensity of E-cadherin in OKC was weak and moderate (P = 0.003). Conclusion: In this study, we observed an increase in the expression of N-cadherin in OKC. In addition, the protein expression levels of E-cadherin in OKC were significantly lower compared to DC and RC. Therefore, it appears that the EMT process likely occurs in OKC and may contribute to its local aggressive behavior.

Dermoid cysts (DMCs) and epidermoid cysts (EDMCs) usually arise in soft tissues, whereas orthokeratinized odontogenic cysts (OOCs) and keratocystic odontogenic tumors (KCOTs) develop in the jaw. In this study, we performed immunohistochemical analysis of cytokeratins (CKs) to examine differences in the lining epithelium of DMCs, EDMCs, OOCs, and KCOTs. In addition, we carried out immunohistochemical examination of langerin to clarify the biological characteristics of the orthokeratinized lining epithelium of DMCs, EDMCs, and OOCs. Seven DMCs, 30 EDMCs, 11 OOCs, and 28 KCOTs were examined immunohistochemically using antibodies against CK10, 13, 14, 16, 17, 19, and langerin. Immunoreactivities for CKs and langerin in oral DMCs and EDMCs were similar to those of lesions affecting the skin. Positive reactivity for CK13 and 17 was evident in OOCs, but not in DMCs/EDMCs. CK10 was significantly positive in all layers except for the basal layer in OOCs, but was negative in KCOTs. CK17 was positive in all layers in KCOTs, and in all layers except for the basal layer in both OOCs and dentigerous cysts. CK19 was negative in OOCs. Langerhans cells were found mainly in OOCs, but were hardly evident in KCOTs. These results suggest that DMCs/EDMCs, OOCs and KCOTs are independent diseases.

Keratocystic odontogenic tumor (KCOT) is a benign tumor that arises sporadically or associated with nevoid basal cell carcinoma syndrome (NBCCS). Its locally aggressive behavior contrasts with its cystic histological appearance. To better understand the interaction between tumor cells and the stroma, the present study aimed to evaluate and compare the immunohistochemical expression of matrix metalloproteinases (MMP-1, -2, and -9), the cellular proliferation index (Ki-67), and the presence of myofibroblasts (MFs) in KCOTs. Eleven cases of isolated KCOT (G1) and 12 cases of KCOT associated with NBCCS (G2) were selected for an immunohistochemical investigation of the proteins MMP-1, MMP-2, MMP-9, Ki-67, and α-smooth muscle actin (α-SMA) in MFs. A group of 6 pericoronal follicles (G3) was included as a normal odontogenic tissue control. Significant differences between the G3 and G1/G2 groups regarding the expression of MMP-1, MMP-9 (in connective tissue), and Ki-67 were observed. In KCOT, there was a positive correlation between the Ki-67 antigen and MMP-1 and between MFs and MMP-1 in the parenchyma. No statistical differences were found between G1 and G2 groups. MMP-1, MMP-9, and proliferative activity appear to play important roles in KCOT pathogenesis. The increased proliferative activity with KCOT was associated with elevated MMP-1 production in the parenchyma, which influenced the growth of the lesion in association with an increased number of MFs.