A large-cohort study of 2971 cases of epulis: focusing on risk factors associated with recurrence.

BACKGROUND: To analyze the clinicopathological features of different histological subtypes of epulis, and evaluate the risk factors associated with recurrence. MATERIALS AND METHODS: A retrospective study including 2971 patients was performed. The patients' sex, age, location, size, histological subtypes, recurrence information, oral hygiene habits, periodontitis symptoms and smoking history were retrieved from the patient medical records and follow-up information. RESULTS: Among the 2971 cases, focal fibrous hyperplasia (FFH) was the most common lesion (60.92%), followed by peripheral ossifying fibroma (POF) (29.32%), pyogenic granuloma (PG) (8.08%) and peripheral giant cell granuloma (PGCG) (1.68%). The peak incidence of epulis was in the third and fourth decade of life, with a mean age of 45.55 years. Female predominance was found in all types of lesions with a female to male ratio of 1.71:1. PG had the highest recurrence rate (17.18%), followed by POF (12.98%), FFH (9.55%) and PGCG (8.82%). Histological subtypes were significantly correlated with the recurrence of epulis (P = 0.013). Regular supportive periodontal therapy (P = 0.050) had a negative correlation with recurrence, whereas symptoms of periodontitis (P < 0.001) had a positive correlation with the recurrence of epulis. CONCLUSIONS: Controlling the periodontal inflammation and regular supportive periodontal therapy might help reduce the recurrence of epulis.

Oral epithelial dysplasia and aphthous ulceration in a patient with ulcerative colitis: a case report.

BACKGROUND: Ulcerative colitis is a chronic inflammatory disease with apparent extraintestinal manifestations, including in the oral cavity. Oral epithelial dysplasia, an exclusive histopathological diagnosis that is used to predict malignant transformation, has never been reported with ulcerative colitis. Herein, we report a case with ulcerative colitis that was diagnosed via extraintestinal manifestations of oral epithelial dysplasia and aphthous ulceration. CASE PRESENTATION: A 52-year-old male suffering from ulcerative colitis came to our hospital complaining of pain on his tongue with a history of 1 week. Clinical examination revealed multiple painful oval ulcers on the ventral surfaces of the tongue. Histopathological examination indicated ulcerative lesion and mild dysplasia in the adjacent epithelium. Direct immunofluorescence demonstrated negative staining along the junction of the epithelium and lamina propria. Immunohistochemical staining with Ki-67, p16, p53 and podoplanin was used to rule out the reactive cellular atypia to inflammation and ulceration of the mucosa. A diagnosis of aphthous ulceration and oral epithelial dysplasia was made. The patient was treated with mouthwash (composed of lidocaine, gentamicin and dexamethasone) and triamcinolone acetonide oral ointment. Oral ulceration healed after one week of treatment. At the 12-month follow-up, minor scarring was observed on the right ventral surface of the tongue, and the patient felt no discomfort in the oral mucosa. CONCLUSION: Oral epithelial dysplasia might also occur in patients with ulcerative colitis despite the low incidence, which should broaden the understanding of oral manifestations of ulcerative colitis.

Adenoid cystic carcinoma in children and young adults: A clinicopathological study of 12 cases.

OBJECTIVE: To investigate the clinicopathological characteristics, immunoprofile, and molecular alterations of adenoid cystic carcinoma (ACC) in children and young adults. MATERIALS AND METHODS: Twelve cases of ACC were included. MYB, MYBL1, Ki-67, type IV Collagen, Laminin, and LAMB1 expression were detected by immunohistochemistry. MYB and MYBL1 rearrangements were detected by fluorescence in situ hybridization. RESULTS: Among 12 patients, four were female and eight were male. Seven cases (58.3%) located in major salivary glands and eight cases (66.7%) were classified as Grade I. Ten tumors (83.3%) had collagenous and hyalinized stroma. MYB was positive in 83.3% cases, and the average Ki-67 labeling index (LI) was 8.3%. LAMB1, type IV Collagen, and Laminin were positive in 91.7%, 66.7%, and 58.3% cases, respectively. Besides, three out of eight tumors had MYB rearrangement. Cases without MYB rearrangement were negative for MYBL1 expression and MYBL1 rearrangement. The average follow-up time was 91.8 months. Four patients had recurrent diseases. CONCLUSIONS: ACC in children and young adults was seen more frequently in males and major salivary glands. Most cases had ECM and hyaline stroma. Grade III tumors, higher Ki-67 LI, negative expression of type IV Collagen, and Laminin showed a tendency of higher recurrence rate.

[The mechanism of scutellarin inhibiting oral leukoplakia carcinogenesis based on network pharmacology and in vitro cytology].

PURPOSE: To investigate the potential mechanisms of scutellarin on oral leukoplakia (OLK) by network pharmacology and further verify by cytology. METHODS: The potential targets of scutellarin acting on OLK were excavated through network pharmacology. PPI network was constructed, and the possible targets and pathways of scutellarin were predicted by GO and KEGG enrichment analysis. CCK-8 and Transwell assays were used to verify the effects of scutellarin on proliferation, migration and invasion of Leuk-1 and Cal-27 cell lines. The expression of related molecules was detected by Western blot to explore potential molecular mechanisms. Statistical analysis was performed with GraphPad Prism 9 software package. RESULTS: There were 29 potential targets of scutellarin acting on OLK, of which HIF-1α was the key target, and the results of GO and KEGG analysis showed that scutellarin was highly involved in the response of cells and tissues to hypoxia and influenced HIF-1 signaling pathway. Scutellarin can significantly inhibit the proliferation (IC50:2 mmol/L), invasion and migration of Leuk-1 and Cal-27 cells(P＜0.05), and downregulated the expression of HIF-1α in Leuk-1 and Cal-27 cells. CONCLUSIONS: Scutellarin can inhibit carcinogenesis of OLK by suppressing HIF-1 signaling pathway.

A pilot study of neoadjuvant combination of anti-PD-1 camrelizumab and VEGFR2 inhibitor apatinib for locally advanced resectable oral squamous cell carcinoma.

Novel neoadjuvant therapy regimens are warranted for oral squamous cell carcinoma (OSCC). In this phase I trial (NCT04393506), 20 patients with locally advanced resectable OSCC receive three cycles of camrelizumab (200 mg, q2w) and apatinib (250 mg, once daily) before surgery. The primary endpoints are safety and major pathological response (MPR, defined as ≤10% residual viable tumour cells). Secondary endpoints include 2-year survival rate and local recurrence rate (not reported due to inadequate follow-up). Exploratory endpoints are the relationships between PD-L1 combined positive score (CPS, defined as the number of PD-L1-stained cells divided by the total number of viable tumour cells, multiplied by 100) and other immunological and genomic biomarkers and response. Neoadjuvant treatment is well-tolerated, and the MPR rate is 40% (8/20), meeting the primary endpoint. All five patients with CPS ˃10 achieve MPR. Post-hoc analysis show 18-month locoregional recurrence and survival rates of 10.5% (95% CI: 0%-24.3%) and 95% (95% CI: 85.4%-100.0%), respectively. Patients achieving MPR show more CD4+ T-cell infiltration than those without MPR (P = 0.02), and decreased CD31 and ɑ-SMA expression levels are observed after neoadjuvant therapy. In conclusion, neoadjuvant camrelizumab and apatinib is safe and yields a promising MPR rate for OSCC.

Orthokeratinized odontogenic cyst (OOC): Clinicopathological and radiological features of a series of 48 cases.

OBJECTIVES: To demonstrate the clinicopathological and radiological features of orthokeratinized odontogenic cysts (OOCs), and analyze the epithelial cell proliferative activity between OOCs and odontogenic keratocysts (OKCs). MATERIALS AND METHODS: Clinicopathological and radiological analyses were performed in all OOC cases. The expression of cell proliferation markers, Ki-67 and cyclin D1, was detected by immunohistochemistry. RESULTS: A total of 48 OOC patients, 28 males and 20 females, were included. The mean age was 33.50 years, with a range of 13-61 years. The mandible was affected five times as frequently as the maxilla (mandible 40, maxilla 8). All OOCs were unilocular radiolucencies with well-defined margins, and 30 of 36 showed loss of continuity of the buccal or lingual cortices based on computed tomography (CT) images. Three cases exhibited root resorption; tooth displacement occurred in 4 cases. The average volume of the cysts on CT was 7794.25 ± 6952.98 mm3. All cysts were treated by enucleation or enucleation after decompression. The average follow-up time was 32.50 ± 27.58 months (ranging from 6 to 65 months), and the overall recurrence rate was 4.44% (2 of 45). Compared with OKCs, Ki-67 and cyclin D1 expression were significantly lower in OOCs (P < 0.001). CONCLUSIONS: OOCs occur more frequently in mandible with a slight male predilection and have a lower proliferative activity than OKCs. Radiologically, OOCs are more likely associated with buccolingual expansion and destruction of cortical bone. Due to the lower aggressiveness and recurrence rate, enucleation or decompression combined with enucleation is the first treatment choice for OOC.

Epithelioid sarcoma: A clinicopathological study of 12 head and neck cases.

OBJECTIVES: To determine the clinicopathological features of epithelioid sarcoma presenting in head and neck region (HNES) and elucidate diagnostic key points and treatment options for HNES. MATERIALS AND METHODS: A total of 12 HNES cases were collected in our department from 2010 to 2020. Their clinical information and pathological features were documented, and relevant follow-up was performed. Immunohistochemistry was carried to analyze the protein markers of HNES. RESULTS: Of the 12 HNES cases, 10 were primary tumors and 2 were metastasized from foot and shoulder, respectively. The patients with primary tumors were significantly younger than those with metastasized ones (22.7 vs 41.5, p = .0157), and male patients outnumbered female patients (3:1). Of all HNES cases, 9 were classic subtype, and 3 were proximal subtype. HNES patients had a poor prognosis, with 5-year overall survival of 41.5% and 5-year relapse-free survival of 22.5%. A loss of INI1 was identified as the hallmark of HNES with 83.3% (10/12) of HNES cases presenting as EZH2 positive. CONCLUSIONS: HNES is more prevalent at younger ages and in males, has a poor prognosis, and exhibits a greater proportion of classic subtype than proximal subtype. EZH2 inhibitor has therapeutic potential in HNES.

Melanotic Neuroectodermal Tumor of Infancy: A Clinicopathological and BRAF V600E Mutation Study of 11 Cases.

PURPOSE: To investigate the clinicopathological features and BRAF V600E mutation of melanotic neuroectodermal tumor of infancy (MNTI). MATERIALS AND METHODS: Eleven cases of MNTI diagnosed at the Department of Oral Pathology were collected. Clinicopathological characteristics were obtained from the medical records. Immunostaining was performed by immunohistochemistry (IHC). Amplification-Refractory Mutation System-qPCR (ARMS-qPCR) and Sanger Sequencing were used to detect BRAF V600E mutation. RESULTS: Of the 11 cases, 3 cases were female and 8 cases were male. The mean age of the first symptoms was 3.2 months (range: 1 to 6 months). Ten cases (90.9%) located in maxilla but only one (9.1%) in mandible. Most of the cases demonstrated well-defined mass with lytic bone destruction and tooth germ affecting radiologically. Histologically, MNTI was consisted of large polygonal melanin-producing epithelioid cells and small round neuroblast-like cells which arranged in irregular alveolar, tubuloglandular and fissured architecture. The epithelioid cells expressed Vim, Pan-CK, NSE and HMB45, while the smalls cells expressed Syn, NSE and scattered Vim. Most cases showed low Ki-67 index (range: <1% to 50%). None of the MNTI cases showed BRAF V600E mutation. Most cases were treated with enucleation (45.4%) or curettage (36.4%). Among the 11 cases, 6 cases had follow-up information, and 2 cases had recurrence lesions after surgery. CONCLUSION: MNTI, an extremely rare tumor, mainly affects male infants with strong preference for maxilla. Distinct histopathological features and immunohistochemical profile are helpful to distinguish from other melanin-containing tumors and small round cell tumors. No BRAF V600E mutation in MNTI is detected in the present study and needs further investigations. The factors that contribute to the local recurrence of MNTI are controversial, but the close follow-up for the patients is recommended.

[Experimental study on the structure and immune cell phenotypes of the lymphoid tissues in oral lichenoid lesions].

PURPOSE: To evaluate the existence of tertiary lymphoid structures(TLS) in oral lichenoid lesions and its compositional characteristics of immune cells. METHODS: Tissue samples of normal oral mucosa, oral lichen planus (OLP) and oral lichenoid tissue reaction(OLTR) were collected, thirty cases in each group. Hematoxylin-eosin(H-E) staining was performed to identify the TLS-like structures, and immunohistochemistry (IHC) staining was applied to assess the structure and amount of infiltrating CD3+ T cells, CD19+, CD20+ B cells, CD21+ follicular dendritic cells (FDC), Bcl-6+ germinal centers, CD34+ PNAd+ venules and CD34+ Gp36+ micro lymphatic vessels in TLS of OLL. Histopathology and molecular markers were used to evaluate the morphological performance of TLS in OLL. Chi-square test (Fisher exact probability method) was applied to compare the proportion of TLS in each group; integral optical density (IOD) method was used to calculate the expression level of each molecular marker, nonparametric t test (Mann-Whitney U test) was employed to analyze their difference. Statistical analysis was performed with GraphPad Prism 7.0 software. RESULTS: In OLP group and OLTR group, 46.7% (14/30) and 23.4% (7/30) cases had TLS-like structures, respectively. The frequency of TLS-like structures was not correlated with the type of disease(P>0.05). Compared with the control group, the molecular markers in OLP group and OLTR group were highly expressed, and the expression of CD19, CD20, and CD21 in OLP group had morphological and structural characteristics of TLS. The expression of Bcl-6(mean and standard deviation of IOD were 15 498±15 108 vs. 1 841±2 276, P<0.0001), CD20 (13 067±9 049 vs. 7 695±5 159, P<0.05), CD21 (13 968±14 560 vs. 2 552±2 584, P<0.0001), PNAd (10 328±10 383 vs. 1 756±1 570, P<0.0001) and Gp36 (12 778±12 390 vs. 2 313±2 578, P<0.0001) showed significant differences between OLP and OLTR tissues, but it could not be used as the criteria for identifying the type of diseases without morphological characters. CONCLUSIONS: TLS exists in OLL lesions, mainly presented as non-classical forms. The classical forms can be occasionally found. CD20 and CD21 can be used as the biomarkers to identify the TLS in OLL. TLS can not be used as the diagnosing criteria for identifying OLP or OLTR.

Analysis of Immunological Characteristics and Genomic Alterations in HPV-Positive Oropharyngeal Squamous Cell Carcinoma Based on PD-L1 Expression.

Programmed death-ligand 1 (PD-L1) expression has been approved as an immune checkpoint inhibitor (ICI) response predictive biomarker; however, the clinicopathological and molecular features of HPV-positive oropharyngeal squamous cell carcinoma [HPV(+)OPSCC] based on PD-L1 expression are not well studied. We aimed to characterize clinicopathological, tumor immune microenvironmental, and molecular features of HPV(+)OPSCC with different PD-L1 expression scored by combined positive score (CPS). A total of 112 cases were collected from 2008-2021 and received PD-L1 and CD8 immunohistochemistry (IHC) staining. 71 samples received DNA sequencing out of which 32 samples received RNA sequencing for immune-related gene alterations or expression analysis. The 32 samples were also subjected to analysis of CD20, CD4, CD8, CD68, Foxp3 and P16 by multiplex immunofluorescence (mIF) staining, and the immune markers were evaluated in the tumor body (TB), tumor margin (TM) and normal stroma (NS) regions separately. Our results showed that of 112 HPV(+)OPSCC tumors, high(CPS≥20), intermediate(1≤CPS<20), and low(CPS<1) PD-L1 expression was seen in 29.5%, 43.8% and 26.8% cases respectively. Non-smoking patients and patients with tumors occurring at the tonsils or having rich lymphocytes infiltration had significantly higher PD-L1 expression. Patients with CPS≥20 had significantly higher tumor mutation burden (TMB, p=0.0058), and PD-L1 expression correlated significantly with CD8+ T cells infiltration, which were ample in tumor regions than in NS in mIF. CD20+, CD4+, CD68+, Foxp3+CD4+ cells were demonstrated to infiltrate higher in TM while CD20+ and CD68+ cells were also enriched in NS and TB regions respectively. However, none of them showed correlations with PD-L1 expression. ARID1A, STK11 alterations were enriched in the low PD-L1 group significantly, while anti-viral immune associated APOBEC mutation signature and immune-related genes expression such as XCL1 and IL11 were positively associated with PD-L1 expression (p<0.05). This is a comprehensive investigation revealing immune and molecular features of HPV(+)OPSCC based on PD-L1 expression. Our study suggested that 73.2% of HPV(+)OPSCC patients may benefit from immunotherapy, and high PD-L1 expression reflects immune-active status of HPV(+)OPSCC accompanied by higher immune effect factors such as TMB, CD8+ cytotoxic T cells and immune-related genomic alterations. Our study offers valuable information for understanding the immune features of HPV(+)OPSCC.

Modification of the anterior-posterior tongue rotation flap for oral tongue reconstruction.

The aim of this study is to evaluate the efficacy and long-term outcome of a modification of the anterior-posterior tongue rotation flap used for reconstruction of tongue following cancer ablation. From April 2016 to September 2017, 21 consecutive patients with tumor classification (T2), lymph node classification (N0) tongue cancer on the anterior and middle one-third portion were selected for this study. A modification of the anterior-posterior tongue rotation flap was selected for tongue reconstruction after the cancer resection. Patient's scorings of swallowing, chewing, speech and taste were recorded. Patient after tongue flap reconstruction acquired satisfied tongue movement that allows normal functional outcomes for swallowing, chewing, speech and taste. Local tumor recurrence was not observed during the follow-up examination (range, 12-24 months). The modification of the anterior-posterior tongue rotation flap was a safe and reliable treatment for medium-size tongue cancer (T2N0), especially suitable to patients who cannot endure prolonged surgery.

Primary pseudomyogenic hemangioendothelioma of right maxilla: a case with immunohistochemistry and FOSB rearrangement study.

OBJECTIVE: Primary pseudomyogenic hemangioendothelioma (PMH) of bone is an extremely rare vascular neoplasm. We present here a case of primary PMH occurring in the maxilla. STUDY DESIGN: A 34-year-old man was referred to our hospital for treatment because of possible recurrence after surgery and chemotherapy of a right maxillary malignant tumor. Morphologic features, immunophenotypes, and FOSB gene rearrangement status of the surgically sectioned sample were assessed by hematoxylin-eosin staining, immunohistochemistry, and fluorescence in situ hybridization, respectively. RESULTS: Morphologically, the tumor cells were arranged in a loose fascicular and sheet-like manner, with a large number of reactive woven bones forming. The most striking feature was the presence of epithelioid cells with abundant brightly eosinophilic cytoplasm, which resembled the rhabdomyoblast in appearance. The tumor was diffusely positive for AE1/AE3, CD31, erythroblast transformation-specific transcription factor, and Friend leukemia integration 1; negative for CD34, CAM5.2, epithelial membrane antigen, and desmin; and had retained expression of integrase interactor 1. The tumor harbored FOSB rearrangement. No distant metastasis was found during the follow-up period (18 months). CONCLUSIONS: To the best of our knowledge, this case represents the first report of PMH arising in the maxilla. The distinct morphologic features, immunophenotypes, and FOSB rearrangement could help achieve precise diagnosis and prevent misdiagnosis of mimics with overlapping features.

Ameloblastoma with mucous cells: A clinicopathological, BRAF mutation, and MAML2 rearrangement study.

OBJECTIVES: To investigate the clinicopathological features, BRAF V600E mutation, and MAML2 rearrangement of ameloblastoma with mucous cell differentiation. MATERIALS AND METHODS: Five cases of ameloblastoma with mucous cell differentiation were retrospectively studied. Clinicopathological features, BRAF V600E mutation, and MAML2 rearrangement were analyzed. Follow-up information was available for all cases. RESULTS: Of five cases, two cases were male and three were female, aged 18-55 years. Four cases were located in the mandible and one case in the maxilla. Histologically, four of the five cases (80%) presented with cystic features and three of the five cases (60%) with varying degrees of squamous metaplasia. The mucous cells were located in the epithelial islands or the luminal aspect of the cystic cavities. The BRAF V600E mutation was found in three of five cases (60%). All the cases showed no MAML2 rearrangement. Two cases were recurrent lesions, and one case had a local recurrence during the follow-up. CONCLUSIONS: Ameloblastoma with mucous cell differentiation is closely related to the cystic features, squamous metaplasia, and shows a high prevalence of BRAF V600E mutation. The absence of MAML2 rearrangement reveals that ameloblastoma with mucous cell differentiation and central mucoepidermoid carcinoma (MEC) are two distinct tumor entities.

[Construction of clinical practice system of oral pathology for long-term program students in stomatology].

PURPOSE: To construct clinical practice system, making up for the shortcomings in the instructional framework of oral histology and pathology, promoting the integration of theory and clinical practice, and enhancing teaching quality of long-term students of stomatology. METHODS: Laying down clinical practice plans and formulating technical operation criteria for pathological experiments, constructing a complete database and training high-level teachers, and evaluating through the following three aspects: practice assessment, teacher-student symposium and questionnaire survey. RESULTS: After completing the clinical practice, the students got the average score of 89.37. In questionnaire survey, the students generally expressed that they had a better understanding of the specialized characteristics and routine skills of the speciality, and improved their practical ability, thus stimulating their interest in self-directed learning. CONCLUSIONS: The construction and practice of clinical practice system of oral histology and pathology can effectively improve the educational objectives and teaching quality of this discipline, which will play a positive role in scientific research and future medical work of long-term students majoring in stomatology.

The MYB/miR-130a/NDRG2 axis modulates tumor proliferation and metastatic potential in salivary adenoid cystic carcinoma.

Increasing evidence has emerged to suggest that N-myc downstream-regulated gene 2 (NDRG2) dysregulation participates in a number of tumor biological processes. However, the role of NDRG2 and miRNA-mediated NDRG2 regulation in salivary adenoid cystic carcinoma (SACC) progression remain unknown. Here, we determined that SACC tissues exhibited decreased level of NDRG2, which was associated with poorer rates of overall survival and distant metastasis-free survival. Silencing NDRG2 promoted SACC cell proliferation and metastasis both in vitro and in vivo. MiRNAs have been reported as vital regulators of NDRG2 expression. Based on micronome sequencing of three paired samples of SACC and normal salivary gland tissue and on an online database analysis, miR-130a was identified as a candidate miRNA that potentially regulates NDRG2. We demonstrated that the expression level of NDRG2 was dramatically reduced by exogenous miR-130a. Moreover, a luciferase assay further validated that miR-130a could degrade NDRG2 mRNA by targeting sites in the NDRG2 3'UTR. A rescue experiment suggested that NDRG2 expression could reverse the miR-130a-mediated promotion of cell proliferation and invasion. The expression of miR-130a has been reported to be regulated by certain transcription factors. In the preset study, we verified that the transcription factor MYB acted as the critical driver in SACC-upregulated miR-130a expression directly and induced NDRG2 downregulation in SACC tissues. Additionally, MYB/miR-130a activated the STAT3 and AKT pathways by downregulating NDRG2. These observations suggest that the MYB/miR-130a/NDRG2 axis, which modulates proliferation and metastasis in SACC, provides promising targets for the treatment of SACC.

Prognostic nomogram for disease-specific survival of carcinoma ex pleomorphic adenoma of the salivary gland.

BACKGROUND: The purpose of this study was to establish an effective prognostic nomogram for carcinoma ex pleomorphic adenoma (Ca-ex-PA) of the salivary gland. METHODS: The nomogram was based on a retrospective study on 223 patients who underwent surgical operation for Ca-ex-PA from 2001 to 2010. The predictive accuracy and discriminative ability of the nomogram were determined by concordance index (C-index) and calibration curve. The results were validated using bootstrap resampling and a prospective study on 111 patients operated on from 2011 to 2012. RESULTS: On multivariate analysis of the primary cohort, independent factors for disease-specific survival (DSS) were age, tumor diameter, degree of capsule invasion, lymph node metastasis, and the interaction between tumor site and histological subtype, which were all selected into the nomogram. The C-index of the nomogram for predicting DSS was 0.90 and 0.86 in the primary cohort and validation cohort, respectively. CONCLUSION: The proposed nomogram resulted in more accurate prognostic predictions for patients with Ca-ex-PA.

Human Papillomavirus 16 Infection and TP53 Mutation: Two Distinct Pathogeneses for Oropharyngeal Squamous Cell Carcinoma in an Eastern Chinese Population.

OBJECTIVES: To investigate the clinicopathological characteristics, human papillomavirus (HPV) infection, p53 expression, and TP53 mutations in oropharyngeal squamous cell carcinoma (OPSCC) and determine their utility as prognostic predictors in a primarily eastern Chinese population. METHODS: The HPV infection status was tested via p16INK4A immunohistochemistry and validated using PCR, reverse blot hybridization and in situ hybridization (ISH) in 188 OPSCC samples. p53 expression levels and TP53 gene mutations were assessed through immunohistochemistry and sequencing, respectively. Clinicopathological characteristics and follow-up information were collected. Overall survival was estimated using the Log-rank test. RESULTS: Overall, 22 of the 188 OPSCC samples were associated with HPV infection. HPV16 was identified in all 22 samples, whereas no samples were positive for HPV18. All 22 HPV-associated OPSCC samples were p53 negative and lacked TP53 mutations. HPV16 positivity, female patients, non-smokers, and patients with histological grade I and stage N0 diseases showed better overall survival (p = 0.009, 0.003, 0.048, 0.009, and 0.004, respectively). No significant differences in overall survival between smoking and non-smoking patients were observed in the HPV-associated OPSCC group. Patients without mutations in TP53 exons 5-8 had better prognoses (p = 0.031) among the 43 sequenced specimens. Multivariate analysis indicated that HPV16 infection status (p = 0.011), histological grade (p = 0.017), and N stage (p = 0.019) were independent prognostic factors for patients with OPSCC. CONCLUSIONS: Distinct from the situation in Europe and America, for the patients with OPSCC in this study, HPV16 infection was relatively low, although it was still the most important independent prognostic predictor for the disease. In addition to the high smoking and drinking rate in this population, HPV16 infection and TP53 dysfunction appear to be two distinct pathogens for OPSCC patients in the eastern Chinese population.

Prognostic factors of carcinoma ex pleomorphic adenoma of the salivary glands, with emphasis on the widely invasive carcinoma: a clinicopathologic analysis of 361 cases in a Chinese population.

OBJECTIVES: To analyze the clinicopathologic characteristics and prognostic factors of salivary carcinoma ex pleomorphic adenoma (Ca-ex-PA) and widely invasive Ca-ex-PA in a Chinese population. STUDY DESIGN: The clinicopathologic parameters of 361 patients with primary Ca-ex-PA from our 2001-2012 cohort were retrospectively reviewed, and the correlation between the parameters and disease-specific survival was statistically analyzed. RESULTS: Of 361 patients with Ca-ex-PA, 229 were male, 132 were female. 191 tumors were invasive carcinoma, 77 minimally invasive, and 93 noninvasive. For 334 (93%) patients, follow-up information was available, and only one of 160 patients with noninvasive or minimally invasive Ca-ex-PA died as a result of the tumor. Of 174 patients with widely invasive Ca-ex-PA, 54 (31%) died as a result of the tumor. Using the Kaplan-Meier method, age, T stage, N stage, histologic grade, and proportion of carcinoma were found to be significantly associated with disease-specific survival of widely invasive Ca-ex-PA. Cox regression analysis indicated that T stage and N stage were independent prognostic factors of disease-specific survival of widely invasive Ca-ex-PA. CONCLUSIONS: Widely invasive Ca-ex-PA had a much worse clinical outcome compared with noninvasive or minimally invasive Ca-ex-PA. T stage and N stage are the most important independent factors for predicting prognosis in Chinese patients with widely invasive Ca-ex-PA.

Low expression of endothelin receptor B (EDNRB) is related to H3K9me3 binding with the EDNRB promoter region and is associated with the clinical T tumor stage in salivary adenoid cystic carcinoma.

OBJECTIVE: To investigate the endothelin receptor B (EDNRB) expression in salivary adenoid cystic carcinoma (ACC) and the mechanism of the regulation of EDNRB expression. STUDY DESIGN: After screening, EDNRB was selected, and the expression was detected using immunohistochemistry in 33 ACC samples (including 6 clinical tumor stage 1 [T1] patients, 13 T2 patients, 9 T3 patients, and 5 T4 patients) and 20 adjacent glands. Interaction between the EDNRB promoter region and histone H3 lysine 9 trimethylation (H3K9me3) was examined using chromatin immunoprecipitation (ChIP) in combination with ChIP-polymerase chain reaction (ChIP-PCR). EDNRB expression in ACC cells treated with chaetocin was detected using quantitative real-time PCR (qRT-PCR) and Western blot tests. RESULTS: EDNRB expression was lower in ACC than that in adjacent glands (P = .006). The expression of EDNRB in patients with advanced T stage was lower than that in patients with early T stage (P = .024). The low EDNRB gene expression group had more H3K9me3 binding regions in the gene promoter (P = .003). EDNRB gene expression significantly increased in the ACC cell lines after treatment with chaetocin. Chaetocin could reduce the interaction between the EDNRB promoter and H3K9me3. CONCLUSIONS: H3K9me3 binding to the EDNRB promoter region could reduce the EDNRB expression. Low EDNRB expression played a role in the progression of ACC tumors.

RASSF1A promoter hypermethylation is a strong biomarker of poor survival in patients with salivary adenoid cystic carcinoma in a Chinese population.

In addition to the clinicopathological parameters, molecular biomarkers are becoming increasingly important in the prognostic evaluation of cancer patients. This study aimed to determine the molecular alterations in the RAS association domain family protein1A gene (RASSF1A) in salivary adenoid cystic carcinoma (ACC) and to evaluate the potential of such alterations as prognostic markers. One hundred and sixty-seven ACC tumor tissues and 50 samples of matched normal salivary gland tissues from the same patients were analyzed for RASSF1A promoter methylation status by bisulfite sequencing PCR (BSP) and/or methylation-specific PCR (MSP). Fifty ACC tumor tissues and matched normal salivary gland tissues were analyzed for loss of heterozygosity (LOH) by examining two microsatellite markers (D3S1478, D3S1621) at 3p21. RASSF1A gene mutations were detected by direct sequencing of all six exons in 50 tumor and normal tissue specimens. Over-all, RASSF1A promoter hypermethylation was detected in 35.3% (59/167) of ACC tissues and was associated with histologically solid tumor pattern (P = 0.002) and advanced TNM stage (P = 0.014). RASSF1A LOH was observed in 18.0% (9/50) of cases, and no somatic mutation of RASSF1A was detected in any cases. RASSF1A promoter methylation was associated with the poor over-all survival (Log-rank test, P <0.001) and disease-free survival (Log-rank test, P <0.001) and identified as an independent predicator of over-all patient survival (P = 0.009) and disease-free survival (P <0.001). It was concluded that RASSF1A methylation is involved in the development, differentiation and progression of ACC and is a strong independent biomarker of poor survival in ACC patients in a Chinese population.