Human Tongue Cancer Cell Line Research Articles

Global analysis of DNA methylation changes during experimented lingual carcinogenesis.

This study aims to assess the role of DNA methylation changes in tongue cancer through a comprehensive analysis of global DNA methylation alterations during experimental lingual carcinogenesis. C57BL/6J mice were subjected to 16-week oral administration of 4-nitroquinoline-1-oxide (4NQO, 50 mg/L). Lingual mucosa samples, being representative of normal tissue (week 0) and early (week 12) and advanced (week 28) tumorigenesis, were harvested for microarray and methylated DNA immunoprecipitation sequencing (MeDIP-Seq). The mRNA and promoter methylation of transforming growth factor-beta-signaling protein 1 (SMAD1) were evaluated with real-time quantitative reverse transcription polymerase chain reaction and Massarray in human lingual mucosa and tongue cancer cell lines. The cytosine guanine island (CGI) methylation level observed at 28 weeks surpassed that of both 12 weeks and 0 weeks. The promoter methylation level at 12 weeks exceeded that at 0 weeks. Notably, 208 differentially expressed genes were negatively correlated to differential methylation in promoters among 0, 12, and 28 weeks. The mRNA of SMAD1 was upregulated, concurrent with a decrease in promoter methylation levels in cell lines compared to normal mucosa. DNA methylation changed during lingual carcinogenesis. Overexpression of SMAD1 was correlated to promoter hypomethylation in tongue cancer cell lines.

Prognostic significance of the FSTL1-DIP2A axis in early-stage tongue cancer.

In tongue cancer, many patients already have metastasis at the time of diagnosis, and such cases are usually unresponsive to treatment, resulting in a poor prognosis. Therefore, there is an urgent need to develop more effective diagnostic and therapeutic methods to cure tongue cancer at the earliest possible stage in clinical practice. Follistatin-like 1 (FSTL1) is known as a negative effector molecule that induces and enhances the refractoriness of cancer cells directly and indirectly via suppressing anti-tumor immunity in various types of cancer. However, the molecular expression, functions, and clinical significance of FSTL1 and its receptor DIP2A in tongue cancer remains to be elucidated. In this study, we revealed that FSTL1, which is highly expressed in tongue cancer cells, plays a key role in its malignancy and is a significant risk factor for recurrence of early-stage tongue cancer. Basic study shows that FSTL1 is abundantly produced from human tongue cancer cell lines, and blocking FSTL1 with specific siRNAs or mAb significantly suppresses cellular functions. Clinical study shows that both FSTL1 and its receptor DIP2A are highly and correlatively expressed in tumor tissues of tongue cancer patients, and high expression levels of both in stage I tumors are significantly associated with shorter relapse-free survival. These suggest that targeting the FSTL1-DIP2A axis may be useful as a biomarker for early prediction of prognosis in tongue cancer patients, and as a therapeutic target for developing new drugs to treat tongue cancer more effectively. This strategy will contribute to improving clinical outcomes in tongue cancer.

Studying the Functional Potential of Ground Ivy (Glechoma hederacea L.) Extract Using an In Vitro Methodology

Glechoma hederacea L., known as ground ivy, has a long history of use in folk medicine. The main bioactive compounds in ground ivy are polyphenolic compounds known for their potent antioxidant and antimicrobial activities and thus have high potential as functional ingredients against bacterial infections and the occurrence of chronic diseases associated with oxidative stress in the human body. The aim of the present study was to determine the biological activity of ground ivy extract on selected human cell lines, including hepatic (HepG2), tongue (CAL 27), gastric (AGS) and colon (Caco-2) cancer cell lines by evaluating cytotoxicity, formation of reactive oxygen species and genotoxicity. The antioxidant capacity of the extract was additionally evaluated using cellular model macromolecules of protein and DNA, bovine serum album and plasmid phiX174 RF1 DNA. The effect of ground ivy extract on representatives of human microflora, including L. plantarum, E. coli and S. aureus, was also studied. The cytotoxicity of the extract depended on the type of cells treated, and the pro-oxidant effect generally decreased with increasing exposure time. The most pronounced genoprotective effect against hydroxyl radical damage was monitored in model plasmid DNA and occurred at the highest tested concentration (0.25 mg mL−1), with 95.89% preservation of the supercoiled form of the plasmid. This concentration also had the most significant antioxidant activity on the model protein—14.01% more than the positive control prepared using Trolox. The ground ivy extract showed high antimicrobial potential against the pathogenic bacteria E. coli and S. aureus.

Effect of Ablative Dose Irradiation on Redistribution and Radioresponse in a Mouse Xenograft Model.

We investigated the effects of ablative dose irradiation on redistribution and radioresponse after the second irradiation in a mouse xenograft model, assuming stereotactic body radiotherapy (SBRT). A human tongue cancer cell line, SAS-Fucci, expressing the fluorescent ubiquitination-based cell cycle indicator (Fucci) that visualizes the cell cycle, was employed in this study. Tumor xenografts formed subcutaneously in nude mice (approximately 6 mm in diameter), with essentially no hypoxic regions, were irradiated at 10 Gy and G2 arrest kinetics were determined using histology sections and a real-time detection method. The second irradiation (10 Gy) was given at intervals of 0 h, 3 h, 1 day, and 4 days after the first irradiation, and tumor regrowth curves were obtained. It was revealed that the ratio of G2-arrested cells showed a much higher peak at 1 day postirradiation compared to 2 Gy, assuming conventional radiotherapy, and gradually decreased thereafter up to 4 days. Tumors irradiated at intervals of 0 h and 1 day demonstrated significantly higher radioresponses than other timings. We conclude that redistribution could contribute to the efficacy of SBRT.

Oral Cancer Cells Release Vesicles that Cause Pain.

Oral cancer pain is attributed to the release from cancers of mediators that sensitize and activate sensory neurons. Intraplantar injection of conditioned media (CM) from human tongue cancer cell line HSC-3 or OSC-20 evokes nociceptive behavior. By contrast, CM from noncancer cell lines, DOK, and HaCaT are non-nociceptive. Pain mediators are carried by extracellular vesicles (EVs) released from cancer cells. Depletion of EVs from cancer cell line CM reverses mechanical allodynia and thermal hyperalgesia. CM from non-nociceptive cell lines become nociceptive when reconstituted with HSC-3 EVs. Two miRNAs (hsa-miR-21-5p and hsa-miR-221-3p) are identified that are present in increased abundance in EVs from HSC-3 and OSC-20 CM compared to HaCaT CM. The miRNA target genes suggest potential involvement in oral cancer pain of the toll like receptor 7 (TLR7) and 8 (TLR8) pathways, as well as signaling through interleukin 6 cytokine family signal transducer receptor (gp130, encoded by IL6ST) and colony stimulating factor receptor (G-CSFR, encoded by CSF3R), Janus kinase and signal transducer and activator of transcription 3 (JAK/STAT3). These studies confirm the recent discovery of the role of cancer EVs in pain and add to the repertoire of algesic and analgesic cancer pain mediators and pathways that contribute to oral cancer pain.

APOPTOSIS MEDIATED ANTI-PROLIFERATIVE ACTIVITY OF KAEMPFEROL AND QUERCETINE ISOLATED FROM THE LEAVES OF ALTINGIA EXCELSA AGAINST HUMAN TONGUE SP-C1 CELL LINES

Introduction: The leaves of Altingia excelsa were found to strongly inhibit SP-C1 human tongue cancer cell lines. This study was focused on identifying antiproliferative compounds found in A. excelsa leaves and assesment their action mechanism. Methods: Extracts of Methanol A. excelsa were fractionated based on their solvent polarity using n-hexane, ethyl acetate, and water. The anti-proliferative testing were tested in vitro against SP-C1 human tongue cancer cell lines using the MTT assay. Isolated the active compound used column chromatography and identified by the spectroscopic method. The isolated compound was also tested for its anti-proliferative testing and ELIZA method of apoptotic induction. Results: This work resulted in the isolation of a flavonoid, which was identified to be kaempferol and quercetin. The compound inhibited SP-C1 proliferation of cell in a time- and dosedependent manner with IC50 values of 0.72 µg/mL and 0. 70 µg/mL for the 24 hours treatments, respectively. Furthermore, promoting apoptosis via the intrinsic pathway, thereby increasing the activity of caspase-8 and caspase-9. Conclusions: These results suggest that kaempferol and quercetin the anticancer compound found in A, excelsa provided a basic for its potential use in cancer disease treatment management

A Study of the Lipidomic Profiles of the CAL-27 and HOK Cell Lines Using EMS Spectra.

ObjectiveThe aim of this study was to explore the lipidomic profiles of the CAL-27 human tongue cancer cell line and the human oral keratinocyte (HOK) cell line.MethodsThe lipidomic differences between the CAL-27 and the HOK cell lines were investigated using non-targeted high-performance liquid chromatography–mass spectrometry lipidomic analysis. The resulting data were then further mined via bioinformatics analysis technology and metabolic pathway analysis was conducted in order to map the most affected metabolites and pathways in the two cell lines.ResultsA total of 711 lipids were identified, including 403 glycerophospholipids (GPs), 147 glycerolipids, and 161 sphingolipids. Comparison of the enhanced MS (EMS) spectra of the two cell lines in positive and negative ionization modes showed the lipid compositions of HOK and CAL-27 cells to be similar. The expressions of most GP species in CAL-27 cells showed an increasing trend as compared with HOK, whereas a significant increase in phosphatidylcholine was observed (p < 0.05). Significant differences in the lipid composition between CAL-27 and HOK cells were shown as a heatmap. Through principal component analysis and orthogonal partial least squares discriminant analysis, noticeably clear separation trends and satisfactory clustering trends between groups of HOK and CAL-27 cells were identified. The numbers of specific lipid metabolites that could distinguish CAL-27 from HOK in positive and negative modes were 100 and 248, respectively. GP metabolism was the most significantly altered lipid metabolic pathway, with 4 metabolites differentially expressed in 39 hit products.ConclusionThis study demonstrated the potential of using untargeted mass spectra and bioinformatics analysis to describe the lipid profiles of HOK and CAL-27 cells.

Kaempferol and Quercetin Isolated from The Leaves of Atingia Excelsa to Arrest Cell Cycle in G0/G1 Phase Human Tongue Cancer SP-C1 Cell Lines

The leaves of Altingia excelsa were found to strongly inhibit SP-C1 human tongue cancer cell lines. This study was focused on identifying the antiproliferative compound found in A. excelsa leaves and assessing its mechanism of action. The active compound was isolated using column chromatography and identified by the spectroscopic method and was also tested for its anti-proliferative properties and the cell cycle analysis in SP-C1 cells by flowcytometry analysis. This work resulted in the isolation of a flavonoid, which was identified to be kaempferol and quercetin. The compounds inhibited SP-C1 cell proliferation in a time- and dose-dependent manner with IC50 values of 0.72 µg/mL and 0. 70 µg/mL for the 24 hours treatments, respectively. Furthermore, the flowcytometry analysis suggested that the compounds exerted its anticancer activities by inhibiting cell cycle. These results suggested that compounds found in A. excelsa providies a basis for its potential use in cancer disease management.&#x0D;

Individual and Combined Treatments with Methylated Resveratrol Analogue DMU-214 and Gefitinib Inhibit Tongue Cancer Cells Growth via Apoptosis Induction and EGFR Inhibition.

The methylated resveratrol analogue 3′-hydroxy-3,4,5,4′-tetramethoxystilbene (DMU-214) has been revealed to exert the anti-cancer activity by a block of the cell cycle at the G2/M phase, apoptosis induction, and metastasis inhibition. These biological events may be involved in crosstalk with the epidermal growth factor receptor (EGFR), which belongs to the ErbB family of receptor tyrosine kinases. Several cancer therapeutic approaches employ small molecules capable of inhibiting tyrosine kinases (e.g., gefitinib). According to more recent reports, combining gefitinib with chemotherapeutics, such as cisplatin, seems to be more effective than monotherapy. The present study aimed to assess the molecular mechanism of the potential anti-proliferative activity of individual and combined treatments with DMU-214 and gefitinib in SCC-25 and CAL-27 human tongue cancer cell lines. We showed for the first time the anti-cancer effects of DMU-214, gefitinib, and their combination in tongue cancer cells triggered via cell cycle arrest, apoptosis induction, and inhibition of the EGFR signaling pathway. The anti-proliferative effects of DMU-214 and gefitinib are also suggested to be related to the EGFR and EGFRP (phosphorylated epidermal growth factor receptor) expression status since we found significantly weaker cytotoxic activity of the compounds tested in SCC-25 cells, which overexpressed EGFR and EGFRP proteins.

Valorization of Banana and Red Beetroot Peels: Determination of Basic Macrocomponent Composition, Application of Novel Extraction Methodology and Assessment of Biological Activity In Vitro

The nutritional and bioactive content of banana and red beetroot peels was investigated. The basic macrocomponent composition was determined using standard AOAC (Association of Official Analytical Chemists) methods, while the recovery efficiency of bioactive compounds was investigated using conventional and innovative extraction techniques (subcritical water extraction, ultrasound- and microwave-assisted extraction). Extracts were analyzed for biological effects in vitro on human hepatic, tongue and colon cancer cell lines. A macrocomponent analysis revealed a notable amount of dietary fiber in banana and beetroot peels (39.0 and 33.6% dmb) and a relatively high content of protein in beetroot peel (18.3% dmb). Regarding the micronutrients-minerals, banana and beetroot peels were shown to be a very good source of potassium (75.06 and 41.86 mg g−1 dmb). Both extracts of banana and beetroot peels obtained by conventional extraction - decoction (100 °C, 20 min) exhibited the highest total phenolic content and antioxidant capacity. Additionally, in banana peel, these extracts were the richest in dopamine content (12.63 mg g−1 dmb). Extraction by infusion (80 °C, 30 min) yielded a beetroot peel extract with the highest total betacyanin content (9.80 mg g−1 dmb). Biological effects in vitro were dose- and time-dependent, as well as influenced by the presence of polysaccharides.

IL-6 Promotes Cell Growth and Is Associated With Poor Prognosis In Patients With Oral Cancer

Abstract Interleukin-6 (IL-6) is a pleiotropic inflammatory cytokine that regulates growth and differentiation of various types of malignant tumors, including oral cancers. Compared to healthy controls, the levels of IL-6 are elevated in sera of patients with oral cancer. In this study, the role of IL-6 in the growth regulation of oral cancer cells was evaluated. The effects of IL-6 on cell growth were evaluated by expressing IL-6 cDNA into the human tongue cancer cell line (SAS), resulting in high-producer clones. In vivo, the effects of tumor-derived IL-6 on growth were studied by using an experimental vaccine model. Two stable clones, consisting of low-producer (IL-6LE), and high-producer (IL-6OE), were subcutaneously inoculated on the flank of each severe combined immune-deficient (SCID) mice. A total of 111 surgically treated HNSCC patients were studied. IL-6 and STAT3 levels were measured by immunohistochemical staining and correlated with clinicopathological variables and patient survival. Treatment of recombinant IL-6 and overexpression of IL-6 by intorducing IL-6 into SAS cells significantly increased clonogenic ability and cell proliferation in vitro compared to the parental SAS cells and the vector controls. In animal model, a statistically significant difference of tumor weight was obtained between the IL-6LE and IL-6OE (*P=0.024) groups in experiment. Clinical data revealed significant relationships between IL-6 and stage*, lymph node metastasis*, recurrence**, disease-free survival* and overall survival* (P &amp;lt; 0.01; *P &amp;lt; 0.05). These findings strongly support the hypothesis that IL-6 acts as a significant autocrine growth factor in vivo and in vivo oral cancers.

Differential properties of mitosis-associated events following CHK1 and WEE1 inhibitor treatments in human tongue carcinoma cells

CHK1 and WEE1 play pivotal roles in G2/M checkpoint following exogenous DNA damage and regulation of DNA replication under normal cellular conditions. Here, we monitored and compared the cell cycle kinetics of mitosis-associated events after CHK1 and WEE1 inhibitor treatments in a human tongue cancer cell line (SAS). A fluorescent ubiquitination-based cell cycle indicator (Fucci) that reflects SCFSKP2 and APCCDH1 E3 ligase activities was used to monitor cell cycle progression. Numerous γH2AX-positive cells were observed within the S phase population of cells following CHK1 inhibitor treatment, and polyploid cells exhibiting DNA damage emerged via abortive mitosis (endomitosis) at 24 h post treatment. While WEE1 inhibitor-treated cells exhibited similar polyploidy via endomitosis at later time points, they possessed fewer γH2AX foci during S phase, and polyploid cells exhibiting DNA damage were scarce. Instead, mitosis duration greatly extended and was accompanied by an abnormal emission of Fucci red fluorescence. Kinetic analysis of Fucci fluorescence revealed that abnormal emission occurred at early M phase in a manner independent of green fluorescence degradation as a marker of APCCDH1 activation. When an inhibitor of the essential spindle checkpoint factor MPS1 was co-treated with a WEE1 inhibitor, the elongated mitosis duration and abnormal red fluorescence were abrogated, and WEE1-induced reduction of clonogenic survival was offset. We demonstrate novel differential effects on mitosis-associated events following CHK1 and WEE1 inhibitor treatments.

Suppression of c-MET overcomes erlotinib resistance in tongue cancer cells.

BackgroundErlotinib is a commonly used molecular-targeted drug for the treatment of tongue cancer. However, the development of acquired resistance to erlotinib hampers its therapeutic use.Materials and methodsTo analyze the erlotinib resistance, long-term and short term survival assay were used to compare the resistance between parental and resistant tongue cancer cells. Flow cytometry, Hochest staining and western blot were used to analyze the apoptosis among the cells. Moreover, Transwell and wound healing assay were used to compare the invasion ability of the cells. To deeply explore the drug resistance in vivo, orthotopic tumor studies were applied. Finally, to explain the mechanism of c-met in erlotinib resistance, shRNA against c-met was used to down-regulate the expression of c-met. And SU11274 also used in orthotopic model.ResultsWe established erlotinib-resistant human tongue cancer cell line by chronic exposure of TCA-8113 cells to increasing concentrations of erlotinib and determined the role of c-MET and EGFR in the development of acquired resistance. We found a significant increase in the phosphorylation of c-MET and an obvious decrease of the phosphorylation of EGFR in erlotinib-resistant cells. Our results also revealed that inhibition of c-MET alone with SU11274 exerted an inhibitory effect on the proliferation of erlotinib-resistant cells in the short term; however, it failed to sustain the inhibitory effect in the long term. Simultaneous inhibition of c-MET and EGFR significantly inhibited the proliferation of erlotinib-resistant cells in both a short and long period. Furthermore, we explored the underlying mechanism and found that treatment of erlotinib-resistant cells with SU11274 or shRNA against c-MET induced the phosphorylation of EGFR. Moreover, our results demonstrated that simultaneous inhibition of c-MET and EGFR significantly inhibited the migration and invasion of erlotinib-resistant cells.ConclusionTaken together, our results suggested that c-MET is involved in acquired drug resistance to erlotinib and that cotargeting of EGFR and c-MET could overcome acquired resistance to erlotinib and inhibit the invasion and metastasis of erlotinib-resistant cells.

Differences between epithelial and mesenchymal human tongue cancer cell lines in experimental metastasis.

Distant metastasis represents the outcome with the worst prognosis for various types of malignant tumors, but little is known regarding the impact of interacting epithelial and mesenchymal phenotypic cancer cells within its etiopathogenesis. In a novel animal model, 48 male athymic Balb/c nude mice underwent subcutaneous and intravenous injection of human tongue cancer cell lines of green fluorescent mesenchymal and red fluorescent epithelial phenotypes, in order to visualize and monitor eventual phenotypic interaction in lung metastasis as well as experimental metastasis in in vivo, ex vivo and histopathological analyses. While the epithelial, but not the mesenchymal, phenotypic human tongue cancer cell line led to direct metastasis in the lungs when injected intravenously, neither of them, even when injected in combination, were able to establish distant metastasis. The results of the present study provide evidence regarding the role of epithelial phenotypic cancer cells in the release of experimental metastasis following tail vein injection in male athymic Balb/c nude mice, in addition to proving fluorescent human tongue cancer cells may be reliably detected under a fluorescence microscope even 8 weeks after the two injection types.

Expression of Plectin-1 and Trichohyalin in Human Tongue Cancer Cells

In basal squamous cells, plectin-1 interacts with intermediate filaments, whereas trichohyalin, which is distributed primarily in the medulla and inner root sheath cells of human hair follicles, plays a role in strengthening cells during keratinization. Although both cytoskeletal proteins occur in trace amounts in human tongue epithelial cells, there are minimal data on their expression in human tongue primary cancer cells. We therefore investigated the expression of plectin-1 and trichohyalin in human tongue epithelial cell line (DOK) and tongue cancer cell line (BICR31) using western blotting and FITC-labeled immunocytochemistry techniques. DOK and BICR31 cells were cultivated to subconfluence in Dulbecco’s Modified Eagle’s Medium containing 0.4 μg/ml of hydrocortisone and 10% fetal bovine serum, and the levels of trichohyalin and plectin-1 were determined by western blot analysis and immunocytochemical staining. Trichohyalin expression was clearly observed, with no differences between DOK and BICR31 cells. Although DOK cells expressed trace levels of plectin-1, obvious plectin-1 bands were detected in western blot analyses of BICR31 cells. Immunocytochemical staining revealed that trichohyalin and plectin-1 localize in the cytoplasm. Trichohyalin was diffusely distributed in both cell lines, and colocalization of trichohyalin and cytokeratin 1/10 was observed in almost all BICR31 cells. There were no correlations between western blot and immunocytochemical data for trichohyalin. Conversely, correlations in immunochemical reactions for plectin-1 were observed. Most DOK cells showed no localization of plectin-1, but strong reactions were detected in the cytoplasm of BICR31 cells. These results indicate that trichohyalin is expressed by cancerous tongue epithelial cells during various stages of malignancy and that plectin-1 provides an index of malignancy.

Long noncoding RNA Lnc‑EGFR promotes cell proliferation and inhibits cell apoptosis via regulating the expression of EGFR in human tongue cancer.

Tongue cancer remains a difficult disease to overcome. Long noncoding RNAs (LncRNAs) have been shown to serve significant roles in the diagnosis and treatment of tongue cancer. Herein, the present study aimed to investigate the role of a newly‑discovered Lnc, Lnc‑EGFR in tongue cancer. The results showed that the transcript level of Lnc‑EGFR was upregulated in patients with tongue cancer and in cultured tongue cancer cell lines. Consistently, expression of EGFR was also elevated selectively in cancerous tissues and malignant cell lines. Knockdown of Lnc‑EGFR inhibited the clonogenic ability and cell viability of human tongue cancer cell lines UM1 and CAL‑27, as evidenced by colony formation assays, and cell proliferation assays. Furthermore, depletion of Lnc‑EGFR in UM1 and CAL‑27 cells increased cell apoptosis by upregulating the activities of caspase‑3, and caspase‑9, but not caspase‑8. Lnc‑EGFR knockdown‑mediated inhibition of clonogenic ability and cell viability was rescued by overexpression of EGFR by adding EGFR recombinant protein into both cell lines. Likewise, Lnc‑EGFR knockdown‑induced cell apoptosis was reversed by co‑treatment with recombinant EGFR protein in UM1 and CAL‑27 cells. All of these results suggested the oncogenic potential of Lnc‑EGFR, which was achieved by positive regulation of EGFR in human tongue cancer.

Folate‑targeted polymeric micelles loaded with superparamagnetic iron oxide as a contrast agent for magnetic resonance imaging of a human tongue cancer cell line.

The aim of the present study was to investigate the feasibility of using folate‑targeted superparamagnetic iron oxide nanoparticles (SPIO) as a magnetic resonance (MR) contrast agent that targets human tongue cancer cells. Folate‑targeted and folate‑free bilayer micelles composed of a diblock copolymer of polyethylene glycol (PEG) and poly‑caprolactone (PCL) that encapsulated SPIO in their hydrophobic core (SPIO‑PEG‑PCL micelles) were synthesized. The cytotoxicity of each set of micelles towards Tca‑8113 cells was examined using methyl thiazolyl tetrazolium (MTT) assays. Tca‑8113 cells were incubated with folate‑free SPIO‑PEG‑PCL micelles and folate‑targeted SPIO‑PEG‑PCL micelles at an Fe concentration of 80µg/ml for 0.5, 1 and 2h. MR imaging was subsequently performed and relative T2 relaxation time was recorded. Endocytosis of each micelle type was observed using Prussian blue staining. The MTT assays demonstrated that varying concentrations of folate‑targeted SPIO‑PEG‑PCL micelles did not result in statistically significant differences in Tca‑8113 cell viability when compared with folate‑free SPIO‑PEG‑PCL micelles (F=0.698; P=0.676). In the MR images obtained, decreased T2‑weighted signal intensity was observed for the folate‑targeted SPIO‑PEG‑PCL and folate‑free SPIO‑PEG‑PCL micelle treatments, particularly after the 2‑h incubation period. However, the folate‑targeted micelles exhibited a significantly greater decrease in signal intensity and a higher relative T2 relaxation time at each time point (P=0.002). In addition, blue intracellular particles were observed in the cells that were incubated with each type of micelle and stained with Prussian blue. However, a greater number of blue particles underwent endocytosis in the folate‑targeted group. Thus, folate‑targeted SPIO‑PEG‑PCL micelles exhibited preferential targeting of Tca‑8113 cells when compared with folate‑free SPIO‑PEG‑PCL micelles, and these results support the potential for these micelles to be used for the early diagnosis of tongue cancer.

Evidence of phenotypic stability after transduction of fluorescent proteins in two human tongue cancer cell lines

ObjectivesThis study investigated the phenotypic stability and biological properties of two human tongue cancer cell lines after transduction of fluorescent proteins. DesignThe human tongue cancer cell lines UM1 and UM2 were cultured with GFP and RFP lentiviral particles stock for 72h. Cells with successful transduction of fluorescent proteins were selected in a medium containing G418 antibiotics for two weeks. The proliferation rates of parental and transduced cell lines were evaluated by their population doubling time (PDT). Transduction efficiency was assessed by fluorescence microscope and flow cytometry. The transduced cells in passage 1, 2, 10, 20 and 30 were collected to check the stability of fluorescent protein expression. Phenotypic stability of the transduced cells was detected by means of cell morphology, cell surface markers and cell function evaluating essay. ResultsThe proliferation rates of the transduced cell lines showed no significant difference compared to their parental cells. Successful transduction with high efficiency (99% up) was demonstrated. High fluorescence expression on both transduced cells was detected until the thirtieth generation. UM1 and UM1-GFP displayed mesenchymal cell characteristics, while UM2 and UM2-RFP cell lines showed properties characteristic of epithelial. ConclusionsTwo human tongue cancer cell lines of epithelial and mesenchymal phenotype respectively, have been successfully labelled with green and red fluorescent proteins. The fluorescence maintained a high expression rate over thirty generations without influencing the original morphological phenotype and cadherin expression.

(±)-Gossypol induces apoptosis and autophagy in head and neck carcinoma cell lines and inhibits the growth of transplanted salivary gland cancer cells in BALB/c mice

Racemic Gossypol [(±)-GOS], composed of both (−)-GOS and (+)-GOS, is a small BH3-mimetic polyphenol derived from cotton seeds. (±)-GOS has been employed and well tolerated by cancer patients. Head and neck carcinoma (HNC) represents one of the most fatal cancers worldwide, and a significant proportion of HNC expresses high levels of antiapoptotic Bcl-2 proteins. In this study, we demonstrate that (±)-GOS inhibits cell proliferation and induces apoptosis and autophagy of human pharynx, tongue, and salivary gland cancer cell lines and of mouse salivary gland cancer cells (SALTO). (±)-GOS was able to: (a) decrease the ErbB2 protein expression; (b) inhibit the phosphorylation of ERK1/2 and AKT; (c) stimulate p38 and JNK1/2 protein phosphorylation. (±)-GOS administration was safe in BALB/c mice and it reduced the growth of transplanted SALTO cells in vivo and prolonged mice median survival. Our results suggest the potential role of (±)-GOS as an antitumor agent in HNC patients.

Radiosensitization of oral tongue squamous cell carcinoma by nanosecond pulsed electric fields (nsPEFs)

Nanosecond pulsed electric fields (nsPEFs) are a non-thermal and non-toxic technology that induce a myriad of biological effects. They have been proven to be effective in tumor shrinkage, but few studies focus on its radiosensitization in oral tongue squamous cell carcinoma. The purpose of this research was to study the radiosensitization effect of nsPEFs on a human oral tongue cancer cell line Tca8113 and to investigate the potential antitumor mechanism. A Tca8113 cell line was tested respectively by MTT assay, clonogenic assay, flow cytometry assay, annexin V-FITC/PI assay, mitochondrial potential assay and total nitric oxide assay. Our results showed that nsPEFs had a time and field strength dependent inhibition effect on Tca8113 cells. The sensitization enhancement ratio (SER) of nsPEFs was 1.453±0.038. Furthermore, radiation induced G2/M arrest was augmented by treatment with nsPEFs. We observed many more Tca8113 cells showing early apoptosis after nsPEFs combined with radiotherapy. Additionally, the NO concentration was significantly increased after nsPEFs treatment. These findings indicate that combination of nsPEFs with radiotherapy can enhance the radiosensitivity of Tca8113 cells and nsPEFs could be a potential radiosensitizer for oral tongue squamous cell carcinoma.