Current Research Status of MicroRNAs in Squamous Cell Carcinoma of the Tongue

Tongue squamous cell carcinoma (TSCC) is the most invasive type of oral malignant tumor, posing a serious threat to human life and health. Its pathogenesis is complex and has a high degree of malignancy. Recurrence and metastasis often lead to poor prognosis. MicroRNAs are a type of single stranded small molecule RNA with only 18 - 25 nucleotides, which can regulate the expression of various genes and participate in the occurrence and development of tumors. Studies have found that microRNA expression profiling can serve as a reliable and stable biological indicator for early diagnosis and prognosis of tumors. This article provides a review of the research status of MicroRNAs in squamous cell carcinoma of the tongue.

Sternocleidomastoid flap for reconstruction of tongue small cell carcinoma: A case report

BACKGROUND The management of tongue carcinoma is excision and radical neck dissection followed with reconstruction.This is a case report of a patient with tongue squamous cell carcinoma(SCC)who underwent the procedure with sternocleidomastoid(SCM)flap reconstruction.CASE SUMMARY A 52-year-old woman without smoking history complained tongue ulcer since 3 years ago.Based on the histopathological examination,the patient was diagnosed with T2N2M0 right tongue SCC and underwent wide excision of tumor;right mandibular;neck dissection and were reconstructed with SCM flap.CONCLUSION SCC of the tongue requires wide excision and dissection of the neck and mandible if infiltration into the surrounding lymph nodes has been found.The SCM flap reconstruction could be used post-surgery.

Molecular signaling in cancer stem cells of tongue squamous cell carcinoma:Therapeutic implications and challenges

Head and neck squamous cell carcinoma is the seventh most common cancer worldwide with high mortality rates.Amongst oral cavity cancers,tongue carcinoma is a very common and aggressive oral cavity carcinoma.Despite the implementation of a multimodality treatment regime including surgical intervention,chemo-radiation as well as targeted therapy,tongue carcinoma shows a poor overall 5-year survival pattern,which is attributed to therapy resistance and recurrence of the disease.The presence of a rare population,i.e.,cancer stem cells(CSCs)within the tumor,are involved in therapy resistance,recurrence,and distant metastasis that results in poor survival patterns.Therapeutic agents targeting CSCs have been in clinical trials,although they are unable to reach into therapy stage which is due to their failure in trials.A more detailed understanding of the CSCs is essential for identifying efficient targets.Molecular signaling pathways,which are differentially regulated in the CSCs,are one of the promising targets to manipulate the CSCs that would provide an improved outcome.In this review,we summarize the current understanding of molecular signaling associated with the maintenance and regulation of CSCs in tongue squamous cell carcinoma in order to emphasize the need of the hour to get a deeper understanding to unravel novel targets.

Effects of hsa-circ-0001862 on Phenotype of Tongue Squamous Cell Carcinoma Cells

[Objectives]To investigate the molecular mechanism of hsa_circ_0001862 on the proliferation,migration,invasion and apoptosis of tongue squamous cell carcinoma Tca-8113 cells.[Methods]hsa_circ_0001862 plasmid was constructed,and the interaction relationship between hsa_circ_0001862 and miR-23a-3p was verified by dual luciferase reporter gene and qRT-PCR.CCK8 assay,colony formation assay,scratch assay and Transwell assay were used to detect the proliferation,migration and invasion ability of Tca-8113 cells.Western blot was used to detect the expression level of apoptosis-related protein molecules.The effects of hsa_circ_0001862 on the apoptosis of Tca-8113 cells was detected.[Results]hsa_circ_0001862 and miR-23a-3p could interact,and their expression was negatively correlated in tongue squamous cell carcinoma cells.In Tca-8113 cells,hsa_circ_0001862 inhibited cell proliferation,migration,and invasion(P<0.01),and promoted cell apoptosis(P<0.01).[Conclusions]The hsa_circ_0001862 interacts with miR-23a-3p,and hsa_circ_0001862 plays an inhibitory role in the development of tongue squamous cell carcinoma.The hsa_circ_0001862 may be a new biomarker and target for the treatment of tongue squamous cell carcinoma.

The Effect of MMP-9 Inhibitors on the Biological Behavior of Human Oral Squamous Cell Carcinoma SCC15 Cell Line Through PI3K/Akt Signaling Pathway

Objective:To investigate the effect of MMP-9 inhibitor(Mki67)on the biology of human oral squamous cell carcinoma SCC15 cell line and to explore its mechanism of action through PI3K/Akt signaling pathway.Methods:SCC15 cells were extracted,and the supernatant was discarded.The cells were then rinsed twice with PBS,and 0,2.5,5,and 10μL of Mki67(50 mg/mL)were added to the culture respectively.The inhibition rate of cell proliferation was detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide(MTT)method,and the cell migration was measured by Transwell chamber test.The cell apoptosis rate was detected by cytometry,and the p-Akt protein content in the cells of each group was determined by a double-antibody sandwich enzyme-linked immunosorbent assay(ELISA)kit.Results:The cell proliferation rates of the 2.5μL,5μL,and 10μL dose groups were all lower than the 0μL group(P<0.05)before treatment,and the cell proliferation rates in the 2.5μL,5μL,and 10μL dose groups decreased overtime(P<0.05).After 24 h,with the increase of Mki67 concentration,the number of migration and invasion gradually decreased(P<0.05),and the number of apoptosis gradually increased(P<0.05);besides,the relative expression of MMP-9,PI3K,and Akt mRNA decreased gradually(P<0.05),and the expression level of Akt mRNA was not statistically significant(P>0.05).Conclusion:MMP-9 inhibitor(Mki67)can inhibit the proliferation and migration of SCC15 cell line and induce apoptosis,and its mechanism of action may be related to the inhibition of PI3K/Akt signaling pathway.

Expression and Significance of IL-6 and bFGF in Human Tongue Squamous Cell Carcinoma

Objective: To detect the expression and pathological significances of Interleukin-6 (IL-6) and basic fibroblast growth factor (bFGF) in tongue squamous cell carcinoma (TSCC). Methods: A tissue array was analyzed immunohistochemically. The expression levels of IL-6 and bFGF in tissues were recorded semi-quantitatively. Results: The positive expression of IL-6 in normal, benign tumor, and carcinoma groups was 12.50% ± 5.575%, 39.00% ± 1.41%, 77.26% ± 17.07% respectively, and the expression among the three groups showed significant differences (P P P Conclusion: It indicates that combining immunohistochemical examination of IL-6 and bFGF has an important reference value on the pathological diagnosis of tongue squamous cell carcinoma.

miR-23a Promoting Cell Proliferation of Human Tongue Squamous Cell Carcinoma Cell through Regulating PPP2R5E

[Objectives]To investigate the mechanism of miR-23a in the proliferation of human tongue squamous cell carcinoma cells.[Methods]Clinical tissue samples of tongue squamous cell carcinoma were collected and Tca8113 and CAL27 were cultured.Real-time quantitative PCR was performed to detect the expressions of miR-23a and PPP2R5E in clinical tissue samples of tongue squamous cell carcinoma.MTT assay,colony formation assay and growth curve assay were used to detect the effect of miR-23a and PPP2R5E on the proliferation of human tongue squamous carcinoma cell.Luciferase reporter assay verified the regulatory relationship between miR-23a and PPP2R5E.[Results]The expression of miR-23a in tongue squamous cell carcinoma was significantly up-regulated(P<0.01).miR-23a promoted the proliferation of tongue squamous cell carcinoma cells.Bioinformatics prediction and luciferin reporting experiments showed that PPP2R5E was a direct target gene of miR-23a.The expression of PPP2R5E was decreased in tongue squamous cell carcinoma.PPP2R5E inhibited the proliferation of tongue squamous cell carcinoma cells.Overexpression of PPP2R5E can reverse the proliferation promoting effect of miR-23a on tongue squamous cell carcinoma cells.[Conclusions]miR-23a can promote the proliferation of tongue squamous cell carcinoma cells through PPP2R5E and miR-23a plays an oncogene role in the occurrence and development of tongue squamous cell carcinoma.

In vitro effect of iASPP on cell growth of oral tongue squamous cell carcinoma

iASPP is an inhibitory member of the apoptosis-stimulating proteins of P53 （ASPP） family. iASPP is over expressed in several malignant tumors and potentially affects cancer progression. However, the expression and potential role of iASPP in oral tongue squamous cell carcinoma （OTSCC） have not been addressed. In our study, we detected iASPP expression in OTSCC by immunohistochemistry, iASPP expression is up-regulated in OTSCC tissues. Moreover, in clinical pathology specimens, we found that increased iASPP expression correlates with poor differentiation and lymph node metastasis. Using multicellular tumor spheroids （MTS） and flow cytometry, we demonstrated that iASPP down-regulation arrests OTSCC cells at the G0/G1 phase, induces OTSCC cell apoptosis and inhibits OTSCC cell proliferation. These results indicate that iASPP plays a significant role in the progression of OTSCC and may serve as a biomarker or therapeutic target for OTSCC patients.

DAPT Enhances the Apoptosis of Human Tongue Carcinoma Cells

Aim To investigate the effect of DAPT （γ-secretase inhibitor） on the growth of human tongue carcinoma cells and to determine the molecular mechanism to enable the potential application of DAPT to the treatment of tongue carcinoma. Methodology Human tongue carcinoma Tca8113 cells were cultured with DAPT. Cell growth was determined using Indigotic Reduction method. The cell cycle and apoptosis were analyzed by flow cytometry. Real-time PCR and Immuno-Fluorescence （IF） were employed to determine the intracellular expression levels. Results DAPT inhibited the growth of human tongue carcinoma Tca8113 cells by inducing G0-G1 cell cycle arrest and apoptosis, The mRNA levels of Hairy/Enhancer of Split-1 （Hes-1）, a target of Notch activation, were reduced by DAPT in a dose-dependent manner. Coincident with this observation, DAPT induced a dose-dependent promotion of constitutive Caspase-3 in Tca8113 cells. Conclusion DAPT may have a therapeutic value for human tongue carcinoma. Moreover, the effects of DAPT in tumor inhibition may arise partly via the modulation of Notch- 1 and Caspase-3.

Role of human papillomavirus in oropharyngeal squamous cell carcinoma: A review

Human papillomavirus(HPV) has been implicated in the pathogenesis of a subset of oropharyngeal squamous cell carcinoma. As a result, traditional paradigms in relation to the management of head and neck squamous cell carcinoma have been changing. Research into HPVrelated oropharyngeal squamous cell carcinoma is rapidly expanding, however many molecular pathological and clinical aspects of the role of HPV remain uncertain and are the subject of ongoing investigation. A detailed search of the literature pertaining to HPV-related oropharyngeal squamous cell carcinoma was performed and information on the topic was gathered. In this article, we present an extensive review of the current literature on the role of HPV in oropharyngeal squamous cell carcinoma, particularly in relation to epidemiology, risk factors, carcinogenesis, biomarkers and clinicalimplications. HPV has been established as a causative agent in oropharyngeal squamous cell carcinoma and biologically active HPV can act as a prognosticator with better overall survival than HPV-negative tumours. A distinct group of younger patients with limited tobacco and alcohol exposure have emerged as characteristic of this HPV-related subset of squamous cell carcinoma of the head and neck. However, the exact molecular mechanisms of carcinogenesis are not completely understood and further studies are needed to assist development of optimal prevention and treatment modalities.

MiR-25-3p attenuates the proliferation of tongue squamous cell carcinoma cell line Tca8113

Objective:To investigate the effects of miR-25-3p on the occurrence,development and proliferation of tongue squamous cell carcinoma cells.Methods:To establish tongue squamous cell carcinoma cell line Tca8113 that stably and highly express miR-25-3p using recombinant reiroviral vector-mediated gene transfer method.The proliferation of transfected Tca8113 was detected by thiazolyl blue tetrazolium bromide(MTT)and cell colony formation assays.eyclnD1,p21^(cipt)and p27^(kipt)mRNA expressions in the transfected Tca-8113 were detected by quantitative PCR.cyclinD1,p21^(cipt),p27^(kipt),AKT,p-AKT,FOXOt and p-FOX01 expressions in the transfected Tca8113 were detected by western blot analysis.In addition,miR-25-3p expression in the tongue squamous cell carcinoma cell line and tissue specimen was also detected by quantitative PCR.Results:Quantitative PCR showed that mitt-25-3p expression in the tongue squamous cell carcinoma cell lines and tissue specimen was significantly lower than that in the adjacent tissue.MTT and cell colony formation assays showed that after miR-25-3p overexpression,the proliferation of transfected Tca8113 was obviously attenuated.Western blot analysis and quantitative PCR showed that after miR-25-3p overexpression.p21^(cipt)and p27^(kipt)expressions were upregulated,while cyclinD1,AKT,FOXO1 expressions were downregulated,and AKT and FOXO1 phosphorylation was inactivated in the transfected Tca8113 cells.Conclusions:MiR-25-3p inhibited the proliferation of tongue squamous cell carcinoma cells and regulated cell cycle-related protein expression,playing an important role in the occurrence and development of squamous cell carcinoma of the tongue.

Evidence of human papilloma virus infection and its epidemiology in esophageal squamous cell carcinoma

瞄准：在食道的有鳞的房间癌(ESCC ) 寻找病毒(HPV ) 感染人的乳突淋瘤的证据并且在河南省移民在食道的癌的致病调查 HPV 感染的潜在的角色和传染病学。方法：乳突淋瘤病毒(PV ) 和 HPV 被 Ultrasensive S-P 免疫组织化学(IHC ) 和原位杂交(ISH ) 在食道的癌纸巾(82 个案例) 和正常粘膜(40 个案例) 决定。结果：当时， IHC 表明 PV 的积极的率分别地是 75.0% ， 68.18% 和 72.5% 积极的率是的 HPV (16/18-E6 )45.0%,36.36% ， 37.5% ，分别地在从在湖北癌症的河南省移民，本地公民和病人的食道的癌组织标本医院。PV 和 HPV (16/18-E6 ) 在所有正常食道的粘膜标本是否定的。没有关联在食道的有鳞的房间癌纸巾并且在等级 1-3 在 HPV 之间被发现食道的有鳞的房间癌房间。原位杂交证明 HPV (16/18 ) DNA 积极的率是 30.0% ， 31.8% ， 25.0% ，分别地在 3 组样品。没有积极杂交信号在 40 个正常食道的粘膜标本被发现。在食道的癌标本的 HPV (16/18 ) DNA 的积极的率在正常粘膜标本(P【0.05 ) 比那显著地高。积极的率不在 3 组食道的癌组织标本(P】0.05 ) 之中是不同的。结论：HPV 感染在在湖北癌症 Hospital.HPV 的河南省移民，本地居民和病人的食道的癌高仔细与食道的有鳞的房间癌被联系。HPV 感染可以在食道的有鳞的房间癌起一个重要作用。

REVERSION OF MALIGNANT PHENOTYPES OF HUMAN LUNG SQUAMOUS CARCINOMA CELLS BY ORNITHINE DECARBOXYLASE ANTISENSE RNA

Abnormally elevated activity of ornithine decarboxylase (ODC), and subsequent polyamine accumulation are intimately associated with the genesis.development and metastasis of cancer. In the present study, to control the growth of tumor cells, ODC antisense RNA was used to transfect human lung squamous carcinoma cell line LTEP-78. Compared with the parental cells, growth of the antisense transfected LTEP-78 cells arrested in G0/Gl phase and colony formation in soft agarose and tumorigenicity in nude mice were significantly reduced. Nucleic acid hybridization demonstrated that the transfectants expressed a high level of ODC antisense RNA and a significantly reduced level of endogenous ODC mRNA.The results suggest that the reversion of malignant phenotypes of human lung squamous carcinoma cells transfected with ODC antisense RNA is associated with the inhibition of polyamine biosynthesis.

Diagnostic value of serum human epididymis protein 4 in esophageal squamous cell carcinoma

BACKGROUND Numerous studies have demonstrated that human epididymis protein 4(HE4)is overexpressed in various malignant tissues including ovarian,endometrial,lung,breast,pancreatic,and gastric cancers.However,no study has examined the diagnostic impact of HE4 in patient with esophageal squamous cell carcinoma(ESCC)until now.AIM To analyze the value of four serum tumor markers for the diagnosis of ESCC,and examine the associations of serum levels of HE4 with ESCC patients’clinicopathological characteristics.METHODS The case group consisted of 80 ESCC patients,which were compared to a control group of 56 patients with benign esophageal disease.Serum levels of HE4,carcinoma embryonic antigen(CEA),alpha fetal protein,and carbohydrate antigen 19-9(CA19-9)were detected by ELISA.The associations of serum HE4 levels with ESCC patients’clinicopathological characteristics such as gender,tumor location,and pathological stage were also examined after operation.RESULTS The result of ELISA showed that serum HE4 level was significantly higher in the patients with ESCC than in the controls,and the staining intensity was inversely correlated with the pathological T and N stages.Serum HE4 levels had a sensitivity of 66.2%and specificity of 78.6%when the cutoff value was set at 3.9 ng/mL.Moreover,the combined HE4 and CA19-9 increased the sensitivity to 83.33%,and interestingly,the combination of HE4 with CEA led to the most powerful sensitivity of 87.5%.Furthermore,A positive correlation was observed between HE4 serum levels and pathological T and N stages(P=0.0002 and 0.0017,respectively),but there was no correlation between HE4 serum levels and ESCC patient gender(P=0.4395)or tumor location(P=0.6777).CONCLUSION The results of this study suggest that detection of serum HE4 levels may be useful in auxiliary diagnosis and evaluation of the progression of ESCC.

OCCULT CERVICAL METASTASIS OF SQUAMOUS CELL CARCINOMA OF TONGUE AMONG CN0 PATIENTS AND ITS TREATMENT

Objective： To explore the treatment of clinically negative neck （CN0） patients with squamous cell carcinoma of the tongue. Methods： 165 CN0 patients with squamous cell carcinoma of the tongue from 1985 to 2002 were investigated retrospectively. Parts of the patients staged at T1, T2 and T3 underwent resection of primary lesion followed by neck observation, and other patients staged above T2 or at T1 but without follow-up were treated with elective neck dissection （END）. All patients were followed up for more than 3 y or until their death. Results： Lymphatic metastasis was identified histologically after operation in 33 of 120 patients treated with END, and 9 of 45 patients treated with resection of primary lesion alone. The overall rate of occult lymphatic metastasis was 25.45%, which increased with the elevating of clinical T stage. The overall rate of neck uncontrolled death was 20.00% for observation group and 5.00% for END group, and significant difference was found between them （P〈0.05）. For T~ patients in the two groups, the rate of neck uncontrolled death was 7.71% and 4.00% respectively, and no significance was found between them （P〉0.05）. When stage T2 and T3 were considered as middle stage together, significant difference （P〈0.05） could be obtained between observation （70.00%） and END group （0%）. Conclusion： The occult metastasis rate of squamous cell carcinoma of tongue increases with the elevating of clinical stage, and elective neck dissection could be considered for NO patients staged over T2 to improve neck control and survival rate; and regional resection alone of primary lesion could be considered for T1N0 patients to improve quality of life if closely followed up is conducted.

Neoneurogenesis in Squamous Cell Carcinoma of Tongue: A New Mechanism for Its Development

Objectives: To explore the role of the growth of new nerves (neo-neurogenesis) in the tumorogenesis of squamous cell carcinoma of tongue. Materials and Methods: 10 formalin-fixed specimens were gained from patients diagnosed with tongue squamous cell carcinoma. Animal models were made by subcutaneous injection in the dorsal midline with Tca-8113 cell line. Mice were sacrificed 2, 4, 6 weeks after cell injection, and tumor tissues were fixed in 4% paraformaldehyde, embedded in paraffin, sectioned. Detection of neo-neurogenesis was stained by Neurofila-ment-L antibody (NF-L) using immunohistochemistry method (IHC) in biopsy from both human body and animal model. Results: IHC staining of NF-L is positive in all 10 paraffins of tongue squamous cell carcinoma sections which suggest that newly formed nerves are observed in tumor microenvironment. NF-L staining is also positive in the paraffins from animal models indicating that the tongue cancer recruits newly formed nerves in its tumorogenesis. Conclusions: Tumor neo-neurogenesis may play an important role in the pathogenesis and development of tongue cancer. From a therapeutic perspective, further studies on the topic may provide new clinical opportunity.

Role of human papillomavirus in the pathogenesis of oral squamous cell carcinoma

Oral cancer is one of the most common cancers and it constitutes a major health problem particularly in developing countries. Oral squamous cell carcinoma(OSCC) represents the most frequent of all oral neoplasms. Several risk factors have been well characterized to be associated with OSCC with substantial evidences. While tobacco and alcohol are the primary risk factors for OSCC development, many epidemiological studies report a strong association with human papillomavirus(HPV) in a subset of OSCC. This article presents our current knowledge on the relationship between HPV and development of OSCC. HPVs are DNA viruses that specifically target the basal cells of the epithelial mucosa. Most experimental data are consistent with the hypothesis that HPV plays a causal role in oral carcinogenesis. Genotypes, such as HPV1 infect epidermal cells, whereas HPV6, 11, 16 and 18 infect epithelial cells of the oral cavity and other mucosal surfaces. Several studies have shown that there is an increased risk of head and neck cancer in the two major HPV 16 oncogenes E6 and E7-positive patients. The presence of antibodies to HPV E6 and E7 proteins was found to be more associated with tumors of the oro-pharynx than of the oral cavity. However, HPV alone appears to be insufficient as the cause of OSCC but requires other co-factors. Although a viral association within a subset of OSCC has been shown, the molecular and histopathological characteristics of these tumors have yet to be clearly defined.

Multimodality functional imaging using DW-MRI and 18F-FDG-PET/CT during radiation therapy for human papillomavirus negative head and neck squamous cell carcinoma: Meixoeiro Hospital of Vigo Experience

AIM To noninvasively investigate tumor cellularity measured using diffusion-weighted magnetic resonance imaging(DW-MRI) and glucose metabolism measured by 18 Flabeled fluorodeoxyglucose positron emission tomography/computed tomography(18F-FDG-PET/CT) during radiation therapy(RT) for human papillomavirus negative(HPV-) head and neck squamous cell carcinoma(HNSCC).METHODS In this prospective study, 6 HPV- HNSCC patients underwent a total of 34 multimodality imaging examinations(DW-MRI at 1.5 T Philips MRI scanner [(n = 24) pre-, during-(2-3 wk), and post-treatment(Tx), and 18F-FDG PET/CT pre- and post-Tx(n = 10)]. All patients received RT. Monoexponential modeling of the DW-MRI data yielded the imaging metric apparent diffusion coefficient(ADC) and the mean of standardized uptake value(SUV) was measured from 18F-FDG PET uptake. All patients had a clinical follow-up as the standard of care and survival status was documented at 1 year.RESULTS There was a strong negative correlation between the mean of pretreatment ADC(ρ =-0.67, P = 0.01) and the pretreatment 18F-FDG PET SUV. The percentage(%) change in delta(?) ADC for primary tumors and neck nodal metastases between pre- and Wk2-3 Tx were as follows: 75.4% and 61.6%, respectively, for the patient with no evidence of disease, 27.5% and 32.7%, respectively, for those patients who were alive with disease, and 26.9% and 7.31%, respectively, for those who were dead with disease.CONCLUSION These results are preliminary in nature and are indicative, and not definitive, trends rendered by the imaging metrics due to the small sample size of HPV- HNSCC patients in a Meixoeiro Hospital of Vigo Experience.

Tinospora crispa extract inhibits MMP-13 and migration of head and neck squamous cell carcinoma cell lines

Objective: To investigate the effect of Tinospora crispa(T. crispa) extract on matrix metalloproteinase 13(MMP-13) expression and cell migration. Methods: The cytotoxicity of T. crispa extract was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on head and neck squamous cell carcinoma(HNSCC) cell lines. The effect on expression of MMP-13 was analysed by RT-PCR and ELISA. The migration was assessed by wound healing assay. Results: MMP-13 m RNA was highly expressed in the metastatic human HNSCC cell lines, HN22 and HSC-3. T. crispa extract at a concentration of 100.0 μg/m L caused about 50% reduction of cell survival. T. crispa extract at a non-toxic concentration of 12.5, 25.0 and 50.0 μg/m L signii cantly suppressed MMP-13 m RNA expression and secreted MMP-13 in both HN22 and HSC-3. The expression of tissue inhibitors of metalloprotease by HSC-3 cells was attenuated by 25.0 and 50.0 μg/m L of T. crispa extract. Addition of the extract to cells in a wound healing assay showed inhibition of cell migration by HN22 cells. Conclusions: These data suggest that T. crispa could be considered as a potential therapeutic drug to prevent metastasis of HNSCC.