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Objective@#To investigate the value of serum MIR4435-2HG level in the diagnosis and prognosis of oral squamous cell carcinoma.@*Methods@#This study was a retrospective case-control study. Five hundred and eighteen samples of oral squamous carcinoma of patients with head and neck squamous cell carcinoma in the cancer genome atlas project (TCGA) database, with long noncoding RNA MIR4435-2HG expression. The median was the boundary, and the patients were divided into high expression group and low expression group, and the 5-year disease-free survival rate and overall survival rate of the two groups were compared. Serum samples from 82 patients with oral squamous cell carcinoma who were admitted to the Department of Oral and Maxillofacial Surgery, Affiliated Stomatology Hospital of Huzhou Univerisity from January 2012 to January 2015 were enrolled to verify the prognostic value of MIR4435-2HG. Bioinformatics is used to predict the biological processes involved in MIR4435-2HG. Use the SPSS 23.0 to set the optimal diagnostic and prognostic cutoff for the MIR4435-2HG.@*Results@#A total of 518 oral squamous carcinoma patients in the TCGA database showed that the 5-year overall survival rate of the MIR4435-2HG high expression group [43.2% (112/259)] was significanthy lower than that of the MIR4435-2HG low expression group [51.7% (134/259)] (P<0.05). The disease-free survival rate of the MIR4435-2HG high expression group [56.8% (147/259)] was significantly lower than that of the MIR4435-2HG low expression group [64.1% (166/259)] (P<0.05). The results of the validation of 82 patients with oral squamous cell carcinoma suggested that the 3-year overall survival rate of the MIR4435-2HG high expression group [40.0% (8/20)] was significantly lower than the MIR4435-2HG low expression group [80.6 % (50/62)] (P<0.05). The clinical and pathological data of serum MIR4435-2HG high expression group and serum MIR4435-2HG low expression group were compared. The results showed that there was no significant difference in gender, age, tumor location and TNM staging between the two groups (P>0.05). The lymph node metastasis rate of the MIR4435-2HG high expression group was significantly higher than that of the low expression group [12.9% (8/62)] (P<0.05). The histological grade of the high expression group [80.0 % (16/20)] was significantly lower than that of the low expression group [24.2 % (15/62)] (χ2=20.030, P<0.05). The results of bioinformatics analysis indicated that the biological functions of MIR4435-2HG target gene were mainly enriched in protein metabolism, processing of rRNA in nucleolus and cytoplasm, SEMA4D induced cell migration process, and mitochondrial translation process.@*Conclusions@#Serum MIR4435-2HG can be used as a potential prognostic marker for oral squamous cell carcinoma.
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Objective@#To investigate the effects of circular RNA hsa_circ_0063772 on the proliferation, migration and invasion of oral squamous cell carcinoma (OSCC) cells.@*Methods@#Thirty-three patients with oral squamous cell carcinoma who were admitted to the Department of Oral and Maxillofacial Surgery, Peking University Shenzhen Hospital from February 2017 to December 2018 were enrolled in this study. Real-time quantative polymerase chain reaction was used to detect the expression level of circular RNA hsa_circ_0063772 in OSCC and corresponding adjacent tissues, OSCC cell lines and human keratinocytes. The expression level of hsa_circ_0063772 was overexpressed in SCC15 and CAL27 cells by using lentivirus, and the effects of this gene on proliferation, migration and invasion of OSCC cells were detected by cell counting assay, scratch assay, Transwell assay, Western blotting and nude mice tumor formation assay.@*Results@#The expression of circular RNA hsa_circ_0063772 in OSCC tissues (9.38±0.34) was lower than that in adjacent tissues (11.30±0.31) (t=5.20, P<0.001), and the expression in OSCC cells (SCC15: 0.12±0.01; SCC25: 0.18±0.02; SCC9: 0.21±0.01; CAL27: 0.13±0.01) was significantly lower than that in human keratinocytes (1.02±0.02) (tSCC15=41.91, tSCC25=29.21, tSCC9=35.16, tCAL27=41.86, P<0.001). Overexpression of hsa_circ_0063772 in SCC15 and CAL27 cells can affect tumor cell proliferation, cell counting assay showed that tumor cell proliferation ability in high expression group (SCC15: 0.76±0.01; CAL27: 0.74±0.02) were lower than empty group (SCC15∶1.22±0.04; CAL27: 0.99±0.03; tSCC15=12.58, tCAL27=6.97; P<0.05). Transwell migration experiment showed the number of migrated cell in high expression group (SCC15∶148.00±14.57; CAL27: 243.00±13.00) were less than empty group (SCC15: 580.30±42.91; CAL27: 424.70±18.66, P<0.01); Transwell invasion assay showed the number of invased cell in high expression group (SCC15: 123.70±6.98; CAL27: 326.00±17.01) were less than empty group (SCC15: 517.70±9.96; CAL27: 454.30±8.09, P<0.01). The results of tumor formation in nude mice showed that the tumor volume and mass of the overexpressed group [(306.40±16.51) mm3; (289.40±11.44) mg] were lower than that of the empty group [(582.60±32.51) mm3, t=7.58, P<0.05; (599.60±21.27) mg, t=7.58, P<0.001].@*Conclusions@#Compared with adjacent tissues, hsa_circ_0063772 is lowly expressed in oral squamous cell carcinoma. High expression of hsa_circ_0063772 can inhibit the proliferation, migration and invasion of OSCC cells.
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BACKGROUND: Good oral health is important for systemic body health and quality of life. Spray oral cleansers are increasingly preferred because of their convenience of carrying and the ease of oral hygiene management. In addition, many kinds of oral cleanser products containing various ingredients with antibacterial, washing, and moisturizing effects are being manufactured. However, concerns about the safety and side effects of oral sprays are increasing, and there is very little information regarding the use and care of oral sprays is available to consumers. This study aimed to investigate the effects of oral spray on oral bacteria and tissue to elucidate the factors that need to be considered when using oral sprays. METHODS: The effects of oral spray on the growth of dental plaque bacteria was assessed using disk diffusion assays. Cytotoxicity and morphological changes in oral epithelial cells were observed by microscopy. The effects of oral spray on dental plaque growth were also confirmed on specimens from permanent incisors of bovines by Coomassie staining. RESULTS: The pH of spray products, such as Perioe Dental Cooling, Cool Sense, and Dentrix, were 3.65, 3.61, and 6.15, respectively. All tested spray products showed strong toxicity to dental plaque bacteria and oral epithelial cells. Compared with those on the control, dental plaque bacteria deposits on the enamel surface increased following the use of oral spray. CONCLUSION: Three types of oral spray, namely Perioe Dental Cooling, Cool Sense, and Dentrix, strongly inhibited the growth of dental plaque bacteria and oral epithelial cells. The oral spray ingredient enhanced dental plaque growth on the enamel surface. Users should be informed of precautions when using oral sprays and the need for oral hygiene after its use.
Subject(s)
Bacteria , Dental Enamel , Dental Plaque , Diffusion , Epithelial Cells , Hydrogen-Ion Concentration , Incisor , Microscopy , Oral Health , Oral Hygiene , Oral Sprays , Plague , Quality of LifeABSTRACT
Objective@#To investigate the effect of long non-coding RNA highly upregulated in liver cancer (lncRNA HULC) on the biological behavior of oral squamous cell carcinoma (OSCC) cell lines.@*Methods@#A total of thirty patients with oral squamous cell carcinoma at Peking University Shenzhen Hospital from March 2017 to March 2018 were enrolled in this study. OSCC and adjacent tissues were extracted during tumor extensive resection. Quantitative real-time PCR (qPCR) was used to detect the expression levels of lncRNA HULC in OSCC and paracancerous tissues and OSCC cell lines. SCC15 and SCC25 cells were transfected with siRNA, and the effects of the gene on the biological behavior of OSCC cells were detected by cell counting assay, scratch assay, Transwell assay and Western blotting.@*Results@#The expression of lncRNA HULC in OSCC tissues (10.98±0.31, n=30) was significantly higher than that in paracancerous tissues (8.39±0.31, n=30) (t=5.93, P<0.001), the expression of lncRNA HULC in OSCC cells (SCC15: 28.58±2.74; SCC25: 16.56±0.87; SCC9: 11.18±1.32; CAL27: 13.92±0.99, n=5) was significantly higher than that in human keratinocytes (1.01±0.00, n=5) (tSCC15=10.08, tSCC25=17.96, tSCC9=7.71, tCAL27=13.09, P<0.001). Down-regulation of lncRNA HULC in SCC15 and SCC25 cells can inhibit the proliferation, migration and invasion of tumor cells and promote tumor cell apoptosis.@*Conclusions@#lncRNA HULC is highly expressed in OSCC and can enhance the proliferation, migration and invasion of cancer cells and inhibit tumor cell apoptosis.
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Objective@#To investigate the effects of sex-detemining region Y box9 (SOX9) expression levels on the proliferation, migration and metastasis in oral squamous cell carcinoma (OSCC).@*Methods@#A total of 74 OSCC pathological specimens were collected from Shanghai OSCC Tissue and Biological Informations Bank, and clinicopathological information of these specimens were collected. Immunohistochemistry assay was used to examine the expression levels of SOX9 in OSCC and to analyze their relationship with clinicopathological features. Cell counting kit-8 assay and cloning formation was used to observe the relationship between the expression levels of SOX9 and the proliferation of OSCC. Transwell experiment and scratch test were used to detect the difference of the ability of OSCC in cell lines with different expression levels of SOX9.@*Results@#The risk of lymph node metastasis in patients with high expression of SOX9 was significantly increased (P=0.010). In the Transwell experiment, the number of HN6 cells (671.0±57.4, P=0.000) migrated to the lower chamber more than that of CAL27 cells (172.0±13.9). In the scratch experiment, HN6 cells [0 h: (93.7±2.1) μm; 6 h: (56.7±2.5) μm; 12 h: (29.7±3.1) μm] migrated faster than CAL27 [0 h: (93.7±1.5) μm; 6 h: (78.0±2.0) μm; 12 h: (42.0±3.0) μm](P<0.05). The migration ability of the cell line (HN6) with high-expression of SOX9 was significantly higher than that in cell line (CAL27) with low-expression SOX9 (P<0.05). The expression levels of SOX9 in OSCC were no significant on cell proliferation (P>0.05).@*Conclusions@#High expression of SOX9 can promote the migration and lymph node metastasis of OSCC. SOX9 is a candidate gene target for the diagnosis and intervention of lymph node metastasis in OSCC.
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Objective@#To explore the expression and clinical significance of circular RNA circHIPK3 in oral squamous cell carcinoma (OSCC), analyze the effect of circHIPK3 on the proliferation of OSCC cells.@*Methods@#The expression of circHIPK3 in OSCC tissues, adjacent non-cancerous tissues and OSCC cell lines were detected by quantitative real-time polymerase chain reaction (qPCR). The correlations between the expression of circHIPK3 in OSCC tissues and the clinicopathological features were analyzed as well. circHIPK3-specific siRNA si-circHIPK3 and negative control siRNA si-NC were designed and synthesised and used to transfect CAL27 and SCC15 cells respectively. The proliferation capacity of CAL27 and SCC15 cells after transfection with si-circHIPK3 was detected by CCK-8 assay. The expression of miR-124 in OSCC was detected by qPCR, and the correlation between expression of circHIPK3 and the expression of miR-124 was analyzed. Using qPCR to detect the expression of miR-124 in CAL27 and SCC15 cells after transfection with si-circHIPK3 and si-NC respectively. Furthermore, using CCK-8 assay to detect the proliferation capacity of CAL27 and SCC15 cells after transfection with si-NC, si-circHIPK3, miR-124 mimic, si-circHIPK3+miR-124 inhibitor.@*Results@#The expression of circHIPK3 in OSCC tissues [2.23 (1.86, 3.00)] was significantly higher than that of the adjacent non-cancerous tissues [1.05 (0.85, 1.26)] (U=1 094, P=0.000). The expression of circHIPK3 in CAL27 (3.02±0.51) and SCC15 cells (3.16±0.75) was higher than those of human normal oral keratinocytes (hNOK) (1.26±0.30) (P=0.000). The expression of circHIPK3 was found to be closely associated with TNM stage (P<0.05) and tumor grades (P<0.05). Knockdown of circHIPK3 can inhibit proliferation of CAL27 and SCC15 cells (P<0.05). The expression of miR-124 in OSCC tissues (0.61±0.35) was significantly lower than that in adjacent non-cancerous tissues (1.13+0.39) (t=-5.36, P<0.05). Correlation analysis showed that the expression of circHIPK3 in OSCC was negatively correlated with the expression of miR-124 (r=-0.767, P<0.001). Moreover, down-regulation of miR-124 rescued the phenotype induced by knockdown of circHIPK3.@*Conclusions@#The expression of circHIPK3 in OSCC was increased, and silencing of circHIPK3 expression can inhibit the proliferation of OSCC cells. Our results suggest that circHIPK3 may play a key role in the occurrence and development process of OSCC through the regulation of miR-124 expression.
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Objective@#To examine the expression and potential clinical significance of CCT (cytidine triphosphate: phosphocholine cytidylyltransferase)-α in oral squamous cell carcinoma (OSCC).@*Methods@#Fifty-eight OSCC and paired adjacent non-malignant epithelia samples (between May 2016 and July 2016) were obtained from dental center, Second Xiangya Hospital, Central South University. CCT-α expression was examined by immunohistochemistry. The relationship between CCT-α and clinicopathological features of OSCC patients was analyzed. Quantitative real-time PCR and Western blot were performed to measure the expression of CCT-α mRNA and protein level in several OSCC cell line and two normal oral epithelial cell line.@*Results@#Immunohistochemistry showed that CCT-α positive staining was found in cell nuclear of OSCC cells and adjacent epithelial cells. CCT-α was positively expressed in OSCC, which was significantly higher than that adjacent to carcinoma tissues (P=0.000). The expression of CCT-α in oral squamous cell carcinoma was correlated with smoking, alcohol consumption, tumor size, differentiation degree and lymph node metastasis. The expression level of CCT-α protein was significantly increased in patients with a history of smoking and alcohol consumption (P=0.001, P=0.004). With the increase of tumor diameter, the expression of CCT-α protein was significantly increased (P=0.005). According to histopathological grade, the lower the degree of tumor differentiation, the higher the expression level of CCT-α protein (P=0.000). The expression of CCT-α protein was significantly higher in patients with lymph node metastasis compared with no lymph node metastasis (P=0.000). Quantitative real-time PCR results showed the CCT-α mRNA expression level was significantly higher in OSCC cells than that in normal oral epithelial cells (P=0.016). The protein expression level of CCT-α was significantly higher in OSCC cells than that in normal oral epithelial cells.@*Conclusions@#CCT-α may play a critical role in the carcinogenesis and development of OSCC.
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Objective@#To investigate the effect of triptolide on human oral cancer cell (HB) proliferation and phosphates and tensin homologue deleted on chromosome ten gene (PTEN) mRNA expression in oral cancer.@*Methods@#The cancer cells were cultured in the medium containing triptolide of different concentrations for 24, 48 and 72 h. Methyl thiazolyl tetrazolium (MTT) method was used to test the rate of growth inhibition of cancer cells, flow cytometer to detect the change of cell cycle and reveres transcription-PCR (RT-PCR) to examine the expression of PTEN mRNA. The expression of PTEN protein was examined by Western blotting.@*Results@#The rate of growth inhibition was (26.92 ± 0.14)%, (38.67 ± 0.11)%, (72.62 ± 0.89)% and (90.42 ± 0.28)%, respectively. The corresponding expression of PTEN mRNA was (3.59±0.21)%, (5.27±0.40)%, (7.18±0.44)% and (9.16±0.50)%, respectively and the corresponding A value of PTEN protein was 0.135±0.007, 0.410±0.020, 0.447±0.017 and 0.884±0.066, respectively. The proportion of G1 phase cells increased from (58.78±0.98)% to (84.13±0.47)%, but the proportion of S phase cells decreased from (25.40±0.43)% to (9.41±0.73)%.@*Conclusions@#The triptolide not only had inhibitory effect on the HB proliferation, but also affected the cell cycle.
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Surgical resection with adequate margins is an essential component of the treatment for patients with oral squamous cell carcinoma (OSCC). A distance of 5 mm or more between healthy tissue to the tumor front is generally accepted as a safe margin. It is very important for surgeons to precisely evaluate the resection area of tumor both pre- and intra-operatively and try to achieve a safe margin, which will result in a decreased risk of local recurrence. The relationship of surgical margin status to patients' prognosis, and factors which will affect surgical margin distance demand are discussed in this paper. We recommend that adequate margins evaluation should take consideration of many factors such as anatomical location, depth of tumor invasion, pattern of tumor invasion, mucosal dysplasia grade and so on. With the development of molecular biology, surgical margin study at molecular level can give us a new strategy to evaluate its adequacy.
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BACKGROUND:RACK1 is strongly associated with the occurrence and development of oral squamous cel carcinoma. However, the occurrence and development of tumor do not depend on a gene or protein, but a long-term complex process of a network structure of multiple genes and multiple molecules, multi-step, multi-stage joint action. Synergism between tumor genes promotes the formation and development of tumor cel s. Therefore, we cannot limit on a single gene or protein to discover the action mechanism of oral squamous cel carcinoma, but should pay attention on signaling network path related to differential protein or gene, investigate the alterations in related protein or gene expression in the whole signaling pathway, and analyze the action mechanism of the interaction of these molecules. OBJECTIVE:To screen differential genes related to oral squamous cel carcinoma, construct an interaction network through bioinformatics using STRING database, and provide clues for future tests. METHODS:In accordance with our previous classic proteomics results and microarray results of oral squamous cel carcinoma, genes with consistent expression and big differences were selected as differential genes. The differential genes were inputted into the database of STRING to find the possible relationship among the protein subunits and to construct network structure of their interaction. RESULTS AND CONCLUSION:The 19 differential proteins of oral squamous cel carcinoma construct a complicated net work, and the differential proteins interact through these networks. GNB2L1-encoded RACK1 is a node protein and interacts with other differential proteins via WD40 repeated protein (number COG2319) andβ-G protein subunit (number KOG0279). WD40 repeated protein (number COG2319) interacts with 5 differential proteins directly and constructs 10 interacting pathways.β-G protein subunit (number KOG0279) interacts with 8 differential proteins directly, which has 11 interacting pathways. We make a network structure picture based on the interaction of these 19 differential genes by the analysis of the STRING database. The results show that the two subunits of RACK1 protein have direct interaction with 8 differential proteins and have 18 interaction pathways on the picture. As a result, RACK1 is the core protein of the network, suggesting RACK1 is the key node protein in oral squamous cel carcinoma.