SLC7A9 as a Potential Biomarker for Lymph Node Metastasis of Esophageal Squamous Cell Carcinoma.

BACKGROUND: The expression of solute carrier (SLC) 7 family genes is reportedly associated with several malignancies. Here, we focused on SLC7A9 and investigated its expression, function, and clinical significance in esophageal squamous cell carcinoma (ESCC). METHODS: SLC7A9 transcription levels were evaluated in 13 ESCC cell lines, and polymerase chain reaction (PCR) array analysis was conducted to detect coordinately expressed genes with SLC7A9. SLC7A9 contributions to proliferation, invasion, and migration were evaluated in ESCC cells subjected to siRNA-mediated gene knockdown and pCMV6-entry plasmid-mediated overexpression. SLC7A9 expression was detected in 189 ESCC tissues by quantitative reverse-transcription (qRT)-PCR and correlated with clinicopathological parameters. RESULTS: The expression levels of SLC7A9 varied widely in ESCC cell lines and correlated with FGFBP1 expression. Knockdown of SLC7A9 significantly suppressed the proliferation, invasion, and migration of the ESCC cell lines. Moreover, overexpression of SLC7A9 enhanced cell proliferation and migration. In analyses of clinical specimens, SLC7A9 mRNA was overexpressed in the ESCC tissues compared with the adjacent normal esophageal tissues. High mRNA expression was significantly associated with high levels of squamous cell carcinoma-related antigen and carcinoembryonic antigen, advanced disease stage, and lymph node metastasis. High SLC7A9 expression was also significantly associated with poor disease-specific and disease-free survival, and lymph node recurrence after radical surgery, but not with the other recurrence patterns. On multivariate analysis, high SLC7A9 expression was an independent predictor of lymph node recurrence. CONCLUSIONS: SLC7A9 influences the malignant behavior of ESCC cells. Tumor SLC7A9 expression may serve as a novel biomarker for predicting lymph node metastasis and recurrence in ESCC patients.

OPLAH Protein Expression Stratifies the Prognosis of Patients With Squamous Cell Carcinoma of the Esophagus.

BACKGROUND/AIM: Squamous cell carcinoma is one of the major subtypes of esophageal carcinoma, and the 5-year overall survival rate of esophageal squamous cell carcinoma patients who underwent curative treatment remains below 40%. We aimed to detect and validate the prognosticators of esophageal squamous cell carcinoma in patients who underwent radical esophagectomy. MATERIALS AND METHODS: Comprehensive analysis of transcriptome and clinical data from The Cancer Genome Atlas revealed OPLAH as one of the differentially expressed genes between esophageal squamous cell carcinoma tissues and normal esophageal mucosa. OPLAH expression changes were significantly associated with a patient prognosis. OPLAH protein levels were further evaluated by immunohisto-chemistry in esophageal squamous cell carcinoma tissues (n=177) as well as in serum samples (n=54) using ELISA. RESULTS: OPLAH mRNA was significantly overexpressed in esophageal squamous cell carcinoma tissues compared to normal esophageal mucosa, and patients with high OPLAH mRNA expression have a significantly poorer prognosis, according to The Cancer Genome Atlas data. The high staining intensity of OPLAH protein in esophageal squamous cell carcinoma tissue clearly stratified patient prognosis. According to multivariable analysis, high OPLAH protein expression was an independent prognostic factor for survival after surgery. Pre-neoadjuvant chemotherapy serum OPLAH protein concentrations were significantly associated with clinical tumor depth and node positivity and, consequently, with advanced clinical stage. The serum OPLAH protein concentration was significantly decreased by neoadjuvant chemotherapy. CONCLUSION: OPLAH protein expression in cancerous tissue and serum may have clinical utility towards stratifying prognosis of patients with esophageal squamous cell carcinoma.

Cytotoxic, genotoxic, and toxicogenomic effects of heated tobacco products and cigarette smoke in human primary keratinocytes.

INTRODUCTION: Heated tobacco products (HTPs) appear to be less harmful to health than conventional cigarettes (CCs). However, limited analytical data are available to support this claim. This study aimed to compare the cytotoxic, genotoxic, and toxicogenomic effects of HTPs and CCs in carcinogenesis via multistep gene mutations in the oral mucosal cells. METHODS: Cigarette smoke extract (CSE) was obtained from HTPs and CCs. Primary human oral keratinocytes (HOKs) were treated with 5% and 20% CSE from HTPs and CCs. Cell survival rate assays were performed after 6, 12, and 24 h. After 6 h, DNA double-strand breaks (DSBs) were evaluated using anti-γH2AX antibodies with immunohistochemistry. mRNAs expressions of mediator of DNA damage checkpoint 1 (MDC1) and ataxia telangiectasia and Rad3-related protein (ATR), were analyzed. Expressions of miR-22 and miR-185 were analyzed because miR-22 targets MDC1 and miR-185, ATR. RESULTS: The HOKs had equivalent survival rates after exposure to the same concentrations of CSE from CCs and HTPs. HTPs increased foci formation of γH2AX in HOKs, as did CCs (without CSE vs 20% HTP, p<0.05; without CSE vs 20% CC, p<0.05). Expressions of MDC1 and ATR decreased in cells exposed to CSE from CCs and HTPs (MDC1: without CSE vs 20% HTP, p<0.05; without CSE vs 20% CC, p<0.05; ATR: without CSE vs 20% HTP, p<0.05; without CSE vs 20% CC, p<0.05). Expressions of miR-22 and miR-185 were not significantly increased when exposed to CSE from CCs or HTPs. CONCLUSIONS: HTPs and CCs had similar cytotoxic effects. HTPs are genotoxic, can cause DSBs, and have toxicogenomic damage because they inhibit the MDC1 and ATR-CHK1 DNA repair pathways in the oral mucosa. The miRNA-mRNA axis was not related to these inhibitions.

Feasibility of oral microbiome profiles associated with oral squamous cell carcinoma.

Objective: Recently, the possibility that oral microbiomes is associated with oral squamous cell carcinoma (OSCC) initiation and progression has attracted attention; however, this association is still unclear. Here, we comprehensively analyze the microbiome profiles of saliva samples using next-generation sequencing followed by determining the association between oral microbiome profiles and OSCC. Materials and Methods: Microbiome profiles in saliva samples from patients with OSCC, oral leukoplakia (OLK), and postoperative OSCC (Post) were analyzed. Candidate OSCC-associated bacteria were identified by comparing the bacterial diversity and relative abundance of each group based on these microbiome profiles, and their applicability as OSCC detection tools were evaluated. Results: There were significant differences in genus abundances (Streptococcus, Aggregatibacter, and Alloprevotella) among the groups from saliva samples. In the OSCC group, compared with the OLK and Post groups, abundances of the genus Fusobacterium, phylum Fusobacteria and phylum Bacteroidetes were markedly increased and that of the genus Streptococcus and phylum Firmicutes were decreased. Conclusion: The results suggested a strong association of these bacteria with OSCC. Especially, phylum Fusobacterium was significantly associated with early recurrence of OSCC. Thus, oral microbiome analysis may have a potential of novel OSCC detection and prognostic tool.

Salivary CCL20 Level as a Biomarker for Oral Squamous Cell Carcinoma.

BACKGROUND/AIM: This study investigated the utility of C-C motif chemokine ligand 20 (CCL20) expression in saliva as a biomarker for oral squamous cell carcinoma (OSCC) and also examined the associated microbiome. MATERIALS AND METHODS: The study group included patients with OSCC or oral potentially malignant disorder (OPMD), and healthy volunteers (HVs). microarray and qRT-PCR were used to compare salivary CCL20 expression levels among groups. Data on CCL20 levels in oral cancer tissues and normal tissues were retrieved from a public database and examined. Furthermore, next-generation sequencing was used to investigate the salivary microbiome. RESULTS: A significant increase in the expression level of CCL20 was observed in both OSCC tissues and saliva from patients with oral cancer. Fusobacterium was identified as the predominant bacteria in OSCC and correlated with CCL20 expression level. OSCC screening based on salivary CCL20 expression enabled successful differentiation between patients with OSCC and HVs. CONCLUSION: CCL20 expression may be a useful biomarker for OSCC.

Salivary CPLANE1 Levels as a Biomarker of Oral Squamous Cell Carcinoma.

BACKGROUND/AIM: This study aimed to identify novel biomarkers for oral squamous cell carcinoma (OSCC) screening to improve the survival rate of patients with oral cancer. MATERIALS AND METHODS: We investigated differential salivary gene expression in patients with OSCC, those with oral potentially malignant disorders (OPMDs), and healthy volunteers (HVs). CPLANE1 was selected for further investigation by microarray analysis. We used quantitative reverse transcription PCR (qRT-PCR) to determine CPLANE1 expression levels in the saliva. The expression of CPLANE1 in normal and oral cancer tissues was analyzed using the Gene Expression database of Normal and Tumor tissues. RESULTS: qRT-PCR analysis of saliva samples showed that CPLANE1 expression levels were significantly higher in OSCC patients than in HVs and OPMDs patients. Furthermore, we developed a screening test for OSCC using CPLANE1 and showed that it had good accuracy. CONCLUSION: Salivary CPLANE1 could be a useful biomarker for OSCC screening and early detection.

A rare case of odontogenic keratocyst extending into the sphenoid bone from the maxilla.

BACKGROUND: Odontogenic keratocyst (OKC) is the third most common odontogenic cyst which arises from cell rests of dental lamina, and usually observed in the jaws. Because OKC is noted for its high rate of recurrence, there are various treatment strategies. Here, we present a rare case of OKC which occupied the entire maxillary sinus and pterygoid process of the sphenoid bone extending nearly to the skull base. CASE PRESENTATION: The patient was a 21-year-old male and underwent surgical removal of the cyst using the Caldwell-Luc procedure which in this case extended the surgical approach to the pterygoid process of the sphenoid bone via the pterygomaxillary junction. However, we found a recurrent lesion in the posterior wall of the maxillary sinus 20 months after the surgery and subsequently performed a secondary cystectomy. Surgical specimens showed positive bcl-2 staining of OKC and negative cytokeratin-10 on immunohistochemistry for both primary and recurrent lesions. CONCLUSION: OKC rarely occurs in the maxillary sinus and extends to the deep maxillary structure and the skull base. In order to prevent recurrence, it is necessary to recognize the exact location of the entire lesion. Careful examination of preoperative CT images is needed to make a complete surgical planning and to perform a reliable surgical procedure.

Management of intraoral bleeding in patients with Waldenström macroglobulinemia: A protocol proposal.

AIM: Waldenström macroglobulinemia is a type of non-Hodgkin lymphoma with poor prognosis observed in patients with hyperviscosity syndrome because of its tendency for fatal symptoms. This study investigated the risk of intraoral bleeding in patients with Waldenström macroglobulinemia based on hyperviscosity syndrome stage and oral health status, and described potential strategies for managing intraoral bleeding. METHODS AND RESULTS: Between April 2012 and March 2017, seven patients with Waldenström macroglobulinemia underwent dental procedures or tooth extraction. Patient records were retrospectively reviewed to obtain data of symptoms, clinical and radiographic findings, treatment details, pretreatment hematological findings, hyperviscosity syndrome status, perioperative method for local haemostasis, postoperative bleeding, and prognosis. The mean patient age was 71.2 years, and the male-to-female ratio was 6:1. Treatment modalities for oral management comprised tooth extraction, scaling, and oral cavity cleaning. Three patients were admitted for hyperviscosity syndrome; one of the patients exhibited postoperative bleeding because of poor oral hygiene, whereas the two other patients with good oral hygiene did not experience intraoral bleeding regardless of the presence of hyperviscosity syndrome. CONCLUSION: We recommended that the risk of oral bleeding in patients with Waldenstrom's macroglobulinemia should be assessed for oral health in addition to the stage of hyperviscosity syndrome.

Salivary NUS1 and RCN1 Levels as Biomarkers for Oral Squamous Cell Carcinoma Diagnosis.

BACKGROUND/AIM: Oral cancer may become advanced because of delay in diagnosis. In order to promote oral cancer screening, simple and highly reliable screening methods that can be implemented at general dental clinics are required. Herein we investigated differential salivary gene expression between oral squamous cell carcinoma (OSCC) patients and healthy volunteers (HV) to identify new biomarkers for OSCC detection. MATERIALS AND METHODS: Candidate genes were selected by microarrays, nuclear undecaprenyl pyrophosphate synthase 1 (NUS1) and reticulocalbin 1 (RCN1) were selected for further investigation. We used real-time quantitative reverse transcription PCR (qRT-PCR) to determine NUS1 and RCN1 expression levels in saliva and tissues. RESULTS: qRT-PCR analysis of clinical samples revealed that OSCC patients had significantly higher expression of salivary NUS1 and RCN1 than HV. CONCLUSION: A combination of NUS1 and RCN1 accurately distinguished patients from controls, and this combination can be implemented as a screening test for OSCC.

Chromobox 2 Expression Predicts Prognosis After Curative Resection of Oesophageal Squamous Cell Carcinoma.

BACKGROUND/AIM: To investigate the function of chromobox 2 (CBX2) in oesophageal squamous cell carcinoma (OSCC). MATERIALS AND METHODS: We used real-time quantitative reverse transcription PCR (qRT-PCR) and immunohistochemistry to determine CBX2 expression levels in 13 human OSCC cell lines and clinical specimens of two independent cohorts of patients with OSCC. RESULTS: PCR array analysis revealed that CBX2 was co-ordinately expressed with WNT5B in OSCC cell lines. RT-qPCR analysis of clinical samples revealed a high tumour-specific CBX2 expression compared with normal oesophageal tissues. High CBX2 expression was significantly associated with shorter disease-specific survival, hematogenous recurrence, and overall recurrence. Analysis of tissue microarrays of one cohort revealed that patients with higher CBX2 levels tended to have a shorter disease-specific survival. CONCLUSION: CBX2 overexpression in OSCC tissues may serve as a novel biomarker for predicting survival and hematogenous recurrence.

Changes in oral microbial profiles associated with oral squamous cell carcinoma vs leukoplakia.

AIM: Oral squamous cell carcinoma (OSCC) is the most frequently occurring cancer among head and neck SCC worldwide. The identification of novel effective biomarkers for early detection may greatly improve the survival rate and prognosis of patients with OSCC. This study aimed to identify specific oral microbial profiles associated with OSCC. METHODS: Saliva samples were collected from oral leukoplakia (OLK) and OSCC patients (N = 6 each) and healthy controls (HC; N = 4). Total bacterial genomic DNA was isolated and 16S rRNA gene survey was performed by next-generation sequencing of the V4 region. The relative distribution of abundance for phylogenetic groups was compared among the OSCC and OLK groups. RESULTS: The 448 operational taxonomic units detected from the libraries were classified into 133 genera, 69 families, 41 orders, 26 classes and 12 phyla. The abundance of phyla Bacteroidetes and genus Solobacterium was notably higher in the OSCC group when compared with the OLK group, whereas those of genus Streptococcus was significantly lower in the OSCC group when compared with the OLK. CONCLUSION: These changes in the salivary microbiome may have potential applications as a novel diagnostic tool for the early detection of OSCC.

Adenoid Cystic Carcinoma With Sialolithiasis of the Left Submandibular Gland: A Case Report and Literature Review.

Adenoid cystic carcinoma is one of the most common salivary gland malignancies with poor long-term prognosis, but the coexistence of sialoliths is extraordinarily rare. In this article, we report a case of 30-year-old woman with a history of submandibular area swelling with intermittent pain increasing during mealtimes that had led her attending physician to diagnose a sialolith in the left submandibular gland on a radiograph 10 years before. However, the surgical specimen proved to be an adenoid cystic carcinoma accompanied with a sialolith. Histopathologically, the submandibular gland was displaced with a fibrous granulation tissue containing a small cribriform carcinoma invading the extracapsular region of the gland. We performed fluorescence in situ hybridization examination with an MYB-NFIB fusion probe of the lesion, with positive results. The patient underwent a supraomohyoid neck dissection as additional procedure because of the possibility of the extracapsular cancer nest remaining around the submandibular gland, but she remains well and disease free 11 years after the first operation.