Laryngeal Cancer Cells Research Articles

Study on the effects of microRNA-203 on the invasion and apoptosis of laryngeal cancer cells via targeting LASP1

Objective: To explore the role of microRNA-203 in laryngeal cancer and its underlying mechanism and clarify the relationship between microRNA-203 and LASP1.Method: microRNA-203 expression in laryngeal cancer tissues and paracancerous tissues was detected by quantitative real time-polymerase chain reaction（qRT-PCR）. The regulatory effects of microRNA-203 on invasion and apoptosis of laryngeal cancer cells were detected by Transwell assay and flow cytometry, respectively. Dual-luciferase reporter gene assay was performed to access the binding condition of microRNA-203 and LASP1. Both mRNA and protein levels of LASP1 in laryngeal cancer cells were detected after transfection with microRNA-203 mimic or microRNA-203 inhibitor by qRT-PCR and Western blot. Rescue experiments were finally performed to detect whether microRNA-203 regulates laryngeal cancer development via targeting LASP1. Result: microRNA-203 was lowly expressed in laryngeal cancer tissues and cell lines.Knockdown of microRNA-203 in Hep-2 cells can promote the invasiveness and inhibit apoptosis of laryngeal cancer cells. Subsequently,LASP1 was predicted to be the target gene of microRNA-203，which was further verified by dual-luciferase reporter gene assay.LASP1 expression was negatively regulated by microRNA-203. Furthermore，rescue experiments showed that microRNA-203 regulates invasion and apoptosis of laryngeal cancer cells via targeting LASP1. Conclusion： Low expression of microRNA-203 could promote the invasion and inhibit apoptosis of laryngeal cancer cells viainhibiting LASP1. microRNA-203 and LASP1 both play a very important role in the development of laryngeal cancer..

Aberrant methylation and downregulation of ZNF667-AS1 and ZNF667 promote the malignant progression of laryngeal squamous cell carcinoma

BackgroundDysregulated long noncoding RNAs (lncRNAs) are involved in the development of tumor. Aberrant methylation is one of the most frequent epigenetic alterations that regulate the expression of genes. The aim of this study was to determine the expression and methylation status of ZNF667-AS1 and ZNF667, elucidate their biological function in the development of LSCC, and identify a cis-regulation of ZNF667-AS1 to ZNF667.MethodsThe expression and methylation status of ZNF667-AS1 and ZNF667 in laryngeal cancer cell lines and LSCC samples were tested respectively. The function of two laryngeal cancer cell lines with overexpression of ZNF667-AS1 or ZNF667 was detected. The regulation between ZNF667-AS1 and ZNF667 was determined.ResultsSignificant downregulation of ZNF667-AS1 was detected in laryngeal cancer cell lines and LSCC tumor tissues. The reduced expression of ZNF667-AS1 was associated with moderate/poor pathological differentiation of LSCC tumor tissues. Aberrant hypermethylation of the CpG sites of ZNF667-AS1, closing to the transcriptional start site (TSS), was more critical for gene silencing, and associated with moderate/poor pathological differentiation. In vitro hypermethylation of promoter region closing to TSS of ZNF667-AS1 decreased the luciferase reporter activity. Overexpression of ZNF667-AS1 reduced the proliferation, migration, and invasion ability of AMC-HN-8 and TU177 cells. The sense strand, ZNF667, was positively correlated with ZNF667-AS1 in expression and function. Overexpression of ZNF667-AS1 led to increased expression of ZNF667 in mRNA and protein level. ZNF667-AS1 and ZNF667 may be associated with epithelial-mesenchymal transition (EMT) process.ConclusionsZNF667-AS1 and ZNF667 are both down-regulated by hypermethylation, and they serve as tumor suppressor genes in LSCC. ZNF667-AS1 regulates the expression of ZNF667 in cis.

Indole-3-Carbinol Inhibits Laryngeal Cancer Growth Through Cell Cycle Arrest

The growth of a variety of tumors are inhibited by indole-3-carbinol (I3C) obviously. But, its role in laryngeal cancer is not clear. The goal of this study was to research the probable roles that laryngeal cancer cell apoptosis and proliferation Hep-2 was influenced by I3C. I3C dose-dependently therapy obviously inhibited Hep-2cell proliferation, and, I3C promoted apoptosis and induced cell morphological changes at 100, 200, 300, 400 μM doses. We discovered that I3C shows anticancer effect through various signal pathways after Hep-2 cells I3C therapy. In Hep-2laryngeal cancer cell line, through decreasing cell cycle-related proteins that include cyclin D1, CDK6, CDK4, and pRb, G1 arrest was induced by I3C. Apart from this, BALB/c nude mice constructed tumor-bearing mouse models. BALB/c nude mice were divided into three groups: treated with I3C, untreated control group and pretreated with I3C. After 8 weeks treatment, the untreated control group developed bigger tumors compared to mice treated or pretreated with I3C, and in the tumors such as cyclin D1, CDK6, CDK4 and pRb cell cycle-related proteins were obviously decreased. Further, the study result showed there was no harmful side effect in the heart, liver and kidney of the I3C-treated nude mice. In conclusion, both in vivo and in vitro I3C inhibited proliferation and induced the Hep-2 cells apoptosis, and showed low toxicity to normal cells. By suppressing the expression of cyclin families and CDK, we deduce that I3C can inhibit the Hep-2 cells growth in vitro. On normal organs and tissues, the I3C had no toxic effects and was safe.

Evaluation of anticancer activity of Melaleuka alternifolia (i. e. tea tree oil) on laryngeal cancer cell line (Hep2) - an in-vitro study

Introduction and Aims A naturally available plant extract of Melaleuka alternifolia tea tree oilTTO is used as a very good antibacterial antifungal antiviral antiprotozoal and anti-inflammatory agent. Currently lot of importance is given for its anticancer effect. Our aim was to evaluate anticancer activity of Melaleuka alternifolia against laryngeal cancer cell line Hep2.Methodology Two different cell lines were used to know the cell viability and cell lysis cytotoxicity in laryngeal cancer Hep2 and monkey kidney cell line Vero used as control. Cell culture The cell lines were maintained in 96 wells micro titer plate containing MEM media supplemented with 10 heat inactivated fetal calf serum FCS containing 5 of mixture of Gentamicin 10ug Penicillin 100 Unitsml and Streptomycin 100microgml in presence of 5 CO2 at 37ordmC for 48-72 hours. Cytotoxicity Assay In vitro growth inhibition effect of test compound was assessed by calorimetric or spectrophotometric determination of conversion of MTT intonbsp Formazan blue by living cells.Results The results represent the mean of five readings. The concentration at which the OD of treated cells was reduced by 50 i.e IC50 value is 0.781microgml of TTO with respect to the untreated control. The standard formula was used for the calculation of cell viability and cell lysis. showed Spearmans Correlation Coefficient Correlation with P value lt0.001 indicating significant results when TTO was treated with Hep2 and Vero cell line. Conclusion TTO is having very good anticancer property against laryngeal cancer cell line Hep2. There was no cytotoxicity noted with Monkey Kidney Vero cell line. The current study can be brought to the level of clinical trials which can show TTO with its greater efficacy related to anticancer activity.

AdoMet triggers apoptosis in head and neck squamous cancer by inducing ER stress and potentiates cell sensitivity to cisplatin.

S-Adenosyl-l-methionine (AdoMet) is a naturally and widely occurring sulfonium compound that plays a primary role in cell metabolism and acts as the principal methyl donor in many methylation reactions. AdoMet also exhibits antiproliferative and proapoptotic activities in different cancer cells. However, the molecular mechanisms underlying the effects exerted by AdoMet have only been partially studied. Inthe current study, we evaluated the antiproliferative effect of AdoMet on Cal-33 oral and JHU-SCC-011 laryngeal squamous cancer cells to define the underlying mechanisms. We demonstrated that AdoMet induced apoptosis in Cal-33 and JHU-SCC-011 cells, involving a caspase-dependent mechanism paralleled by an increased Bax/Bcl-2 ratio. Moreover, we showed, for the first time, that AdoMet induced ER-stress in Cal-33 cells and activated the unfolded protein response, which can be responsible for apoptosis induction through the activation of CHOP and JNK. In addition, AdoMet-induced ER-stress was followed by autophagy with a consistent increase in the levels of the autophagic marker LC3B-II, which was indeed potentiated by the autophago-lysosome inhibitor chloroquine. As both escape from apoptosis and decreased activation of JNK are mechanisms of resistance to cisplatin (cDPP), an agent usually used in cancer therapy, we have evaluated the effects of AdoMet in combination with cDPP on Cal-33 cells. Our data showed that the combined treatment resulted in a strong synergism in inhibiting cell proliferation and in enhancing apoptosis via intrinsic mechanism. These results demonstrate that AdoMet has ER-stress-mediated antiproliferative activity and synergizes with cDDP on cell growth inhibition, thus providing the basis for its use in new anticancer strategies.

MicroRNA-384 regulates cell proliferation and apoptosis through directly targeting WISP1 in laryngeal cancer.

Laryngeal cancer (LC) is an increasingly common malignant tumors of head and neck cancer. Aberrant expression of microRNA (miRNA) is closely related with LC development. In the current study, we investigated the biological function and underlying molecular mechanism of miR-384 in LC. The results showed that the miR-384 expression was markedly downregulated in LC tissue and cell lines (TU212 and TU686) as compared with that of adjacent nontumor tissues and a normal human bronchial epithelial cell line. Next, we performed gain-of-function and loss-of-function experiments in the TU212 and TU686 cells by transfecting the cells with miR-384 mimics, miR-384 inhibitor, or miRNA control. Moreover, results showed that miR-384 mimic remarkably inhibited LC cell proliferation, which was notably decreased by miR-384 inhibitor. Furthermore, miR-384 mimics notably increased the amounts of DNA fragmentation from the apoptotic cells (a hallmark of apoptosis) and the caspase-3 activity, whereas miR-384 inhibitor resulted in a decline of DNA fragmentation and the caspase-3 activity compared with its control. In addition, a dual-luciferase reporter assay confirmed that Wnt-induced secreted protein-1 (WISP1) gene was a direct target of miR-384. MiR-384 mimic remarkably inhibited the messenger RNA and protein expression of WISP1, which was upregulated by miR-384 inhibitor as compared to its control. WISP1 knockdown by small interfering RNA inhibited LC cell proliferation and promoted cell apoptosis. WISP1 overexpression partly abrogates the effect of miR-384 overexpression. Taken together, these data indicate that miR-384 regulates LC cell proliferation and apoptosis through targeting WISP1 signaling pathway, providing a novel insight into the LC treatment.

Coordination Compounds of Cobalt(II), Nickel(II), and Copper(II) Halides with 2-Methyl-1,2,4-Triazolo[1,5-a]benzimidazole

New coordination compounds of Co(II), Ni(II), and Cu(II) halides with 2-methyl-1,2,4-triazolo[1,5-a]benzimidazole (L), [Co(L)2Cl2] (Ia), [Ni(L)2(H2O)4]Cl2 · 4H2O (II), and [Сu2(L)4(µ-Br)2Br2] (IIIa), are synthesized. The complexes are studied by X-ray structure analysis (СIF files CCDC nos. 1825006–1825008), electronic (diffuse reflectance) spectroscopy, and IR spectroscopy. In compounds Ia, II, and IIIa, ligand L is coordinated to the metal by the N4 atom of the triazole fragment. The study of the cytotoxic effect of the ligand and synthesized complexes on the Hep-2 line (laryngeal cancer cells) shows a considerably higher activity of the copper(II) complex than that of the ligand.

Anticancer activity of saponin isolated from Albizia lebbeck using various in vitro models

Ethnopharmacological relevanceAlbizia lebbeck (L.) Benth. (Family: Mimosaceae) is commonly known as Sirisha in Sanskrit. The leaves and pods of A. lebbeck were claimed to be used against cancer in traditional medicine. Previous studies using bark, leaves, seeds and pods of A. lebbeck showed cytotoxic activity against hepatic, colon, larynx, cervical and breast cancer cell lines. Aim of the studyTo evaluate the anticancer activity of saponin rich fraction of Albizia lebbeck by using various in vitro models. Materials and methodsAlbiziasaponins (A-E) are oleanene triterpene presents in Albizia lebbeck were used for in silico studies. In silico testing of albiziasaponins for structure based pharmacological activity prediction using PASS Online software and docking with Autodock tool and Autodock Vina revels it's anticancer and apoptogenic potential. Antiproliferative activity of saponin rich fraction of A. lebbeck was performed using MCF-7 human breast cancer cells by MTT assay methods. Anti-angiogenic property of saponin rich fraction of A. lebbeck was evaluated in in vitro shell less chick embryo cultures with different concentrations (0.1 µg/ml, 0.5 µg/ml, and 1 µg/ml) by using ImageJ software. In vitro cultured lymphocytes chromosomal aberration assay was performed to determine the physical integrity of chromosomes in cells and effect of saponin rich fraction of A. lebbeck on cell cycle. Apoptogenic potential was evaluated using Caspases-3 and Caspase-8 ELISA assay in MCF-7 cells. ResultsResult of MTT assay showed IC50 of saponin rich fraction of A. lebbeck at 1 μg/ml in MCF-7 cells. Treatment with saponin rich fraction of A. lebbeck significantly (p < 0.05) reduced angiogenic parameters. Significant chromosomal aberrations (hypodiploid, hyperdiploid, ring, premature separation, Dicentric fragments, Acentric fragment, chromatid break, and chromosomal gap) were observed in saponin rich fraction of A. lebbeck treated groups. Treatment with saponin rich fraction of A. lebbeck increased levels of Caspases-3 (optical density of 0.24 at 450 nm) and Caspase-8 (optical density of 0.31 at 450 nm) as compared to staurosporine (optical density of 2.47 and 2.65 for caspases-3 and -8 respectively at 450 nm). ConclusionIn our study, saponin rich fraction of A. lebbeck showed antiproliferative, antiangiogenic and apoptogenic potential using various in-vitro models. It also found to increase chromosomal aberration and thereby may affect cell cycle.

Anticancer Activity of Mukonal Against Human Laryngeal Cancer Cells Involves Apoptosis, Cell Cycle Arrest, and Inhibition of PI3K/AKT and MEK/ERK Signalling Pathways.

BackgroundLaryngeal cancer is one of the major malignancies of the neck and head and is responsible for considerable mortality across the globe. The treatments for laryngeal cancer mainly involve surgical interventions followed by chemotherapy. However, due to unsatisfactory results, constant relapses and the adverse effects associated with the currently used drugs, there is pressing need to develop effective drug options for treatment of laryngeal cancer. Therefore, this study was undertaken to investigate the anticancer effects of a plant-derived alkaloid, Mukonal, against human AMC-HN-8 laryngeal cancer cells.Material/MethodsThe WST-1 and clonogenic assays were employed to determine the cell viability. Apoptosis was detected by Hoechst and AO/EB staining. Cell migration and cell cycle analysis was performed by Transwell assay and flow cytometry, respectively. Protein expression was examined by Western blotting.ResultsThe results revealed that Mukonal reduced the viability of laryngeal cancer cells dose-dependently. The IC50 of Mukonal was found to be 10 μM. However, the effects of Mukonal on the normal HuLa-PC cells was found to be 140 μM. The decrease in the viability of the AMC-HN-8 laryngeal cancer cells was found to be due to the induction of apoptosis and G2/M cell cycle arrest. Mukonal also suppressed the cell migration and of the AMC-HN-8 laryngeal cancer cells. Mukonal could also inhibit the PI3K/AKT and MEK/ERK signalling pathways in a concentration-dependent manner.ConclusionsTaken together, we conclude that Mukonal could prove a beneficial lead molecule for the treatment of laryngeal cancer.

Hydrogen peroxide-induced mitophagy contributes to laryngeal cancer cells survival via the upregulation of FUNDC1.

The purpose of our study was to investigate an underlying mechanism that hydrogen peroxide-induced mitophagy contributed to laryngeal cancer cells survivals under oxidative stress condition. Tumor tissue and serum samples were collected from patients with laryngeal cancer. The Hep2 cell, a human laryngeal carcinoma cell, was used in in vitro experiments. The levels of lipid peroxidation were analyzed by ELISA. Knockdown of FUNDC1 was performed by RNAi. The changes of target proteins were determined by qRT-PCR and western blot. The cells were analyzed for changes in proliferation using cell counting kit-8 and mitophagy by the mitochondrial membrane potential assay and transmission electron microscopy. FUNDC1 in laryngeal cancer tissues were relative to the levels of lipid peroxidation in laryngeal cancer patients, which suggested that FUNDC1 was associated with the status of oxidative stress in the laryngeal cancer patients. Hydrogen peroxide significantly induced the elevation of FUNDC1, a mitophagic factor, in a time- and dose-dependent manner in laryngeal cancer cells, which was dependent on ERK signal activation. Knockdown of FUNDC1 by the siRNA attenuated the survival of laryngeal cancer cells under hydrogen peroxide stimulation. Moreover, the elevated FUNDC1 was required for the occurrence of mitophagy under hydrogen peroxide stimulation, which was identified by transmission electron microscopy, the alterations of mitochondrial permeability transition and the specific mitochondrial protein, hsp60. Inhibition of mitophagy with cyclosporine A could also effectively attenuate the laryngeal cancer cells survival under hydrogen peroxide stimulation. Hydrogen peroxide upregulated the expression of FUNDC1 through the activation of ERK1/2 signal to trigger a mitophagic response, giving laryngeal cancer cells a befit for survival. These findings suggested that FUNDC1 might be a potential target for the treatment of laryngeal cancer accompanied with high lipid peroxidation status.

MicroRNA-203 inhibits invasion and induces apoptosis of laryngeal cancer cells via targeting LASP1.

To explore the role of microRNA-203 in laryngeal cancer and its underlying mechanism in regulating cell invasion and apoptosis. MicroRNA-203 expression in laryngeal cancer tissues and paracancerous tissues was detected by quantitative real time-polymerase chain reaction (qRT-PCR). The regulatory effects of microRNA-203 on the invasion and apoptosis of laryngeal cancer cells were detected by transwell assay and flow cytometry, respectively. Dual-Luciferase reporter gene assay was performed to access the binding condition of microRNA-203 and LASP1. Both mRNA and protein levels of LASP1 in laryngeal cancer cells were detected after transfection with microRNA-203 mimic or microRNA-203 inhibitor by qRT-PCR and Western blot, respectively. Rescue experiments were finally performed to detect whether microRNA-203 regulates laryngeal cancer development via targeting LASP1. MicroRNA-203 was lowly expressed in laryngeal cancer tissues and cell lines. MicroRNA-203 knockdown in Hep-2 cells can promote the invasion and inhibit the apoptosis of laryngeal cancer cells. Subsequently, LASP1 was predicted to be the target gene of microRNA-203, which was further verified by the Dual-Luciferase reporter gene assay. LASP1 expression was negatively regulated by microRNA-203. Furthermore, rescue experiments showed that the regulatory effects of microRNA-203 on the invasion and apoptosis of laryngeal cancer cells were reversed by LASP1. We showed that lowly expressed microRNA-203 could promote the invasion and inhibit apoptosis of laryngeal cancer cells via inhibiting LASP1.

Hesperidin inhibits angiogenesis, induces apoptosis, and suppresses laryngeal cancer cell metastasis

Introduction: Laryngeal carcinoma is the most common malignant head-and-neck tumor. Due to the low survival rate and the inadequate response to chemotherapy, effective therapy remains a challenge. Objectives: Therefore, the identification of new therapeutic options that preserve the larynx is needed. Hesperidin (Hsp) is a nontoxic plant flavanone that has proven effective against cancer. Materials and methods: Hence, the current in vivo and in vitro study was conducted to determine whether Hsp might suppress metastasis of cancer larynx. Results: In an in vivo mouse metastasis model, Hsp suppressed metastasis of human Hep2 laryngeal cancer cells to the livers and lungs. In vitro assays, Hsp significantly inhibited angiopoietin 1 secretion (an angiogenic promotor) and increased annexin-V (an apoptotic indicator) in Hep2 cell culture at relatively low levels (10 μM). Conclusions: These studies suggest that Hsp deserves further investigation as a possible treatment option for laryngeal cancer.

Tanshinone sensitized the antitumor effects of irradiation on laryngeal cancer via JNK pathway.

Laryngeal cancer is a common cancer occurred in the head and neck. Irradiation sensitivity is a problem affecting the treatment of laryngeal cancer. Tanshinone IIA has been reported to play an important role in treating multiple diseases; yet, whether Tanshinone IIA can be an irradiation sensitizer has not been reported. Clonogenic assay, annexin‐V/propidium iodide double‐staining assay, and Cell Counting Kit‐8 assay were performed to detect cell survival, proliferation, apoptosis, and viability. Mouse laryngeal cancer xenograft model was established and subjected to tumor size analysis. Tanshinone IIA treatment increased the irradiation sensitivity of laryngeal cancer cells by reducing cell survival, viability and proliferation, and increasing cell apoptosis. Tanshinone IIA treatment increased the survival period of mice in the in vivo laryngeal cancer model, evidenced by decreased growth and weight of tumors, which was possibly mediated through the JNK pathway. Tanshinone IIA increases the sensitivity to irradiation in laryngeal cancer cells and in vivo laryngeal cancer model, suggesting that Tanshinone IIA can be a therapeutic antitumor agent for treating laryngeal cancer.

MiR-140-5p inhibits larynx carcinoma invasion and angiogenesis by targeting VEGF-A.

Larynx carcinoma is a common head-neck malignant tumor. Recent investigations showed the involvement of microRNA (miR) in regulation of multiple tumors. miR-140-5p showed decreased expression in various cancers, but without knowledge regarding its expression in larynx carcinoma and effects on cell invasion and angiogenesis. Real-time quantitative PCR was firstly employed to measure miR-140-5p expression in larynx carcinoma and controlled tumor adjacent tissues. In larynx cancer cell line, agomir or antagomir of miR-140-5p was applied to up-regulate or down-regulate miR-140-5p, respectively. Western blot was used to evaluate vascular endothelial growth factor A (VEGF-A) expression, and cell proliferation was modified by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H- tetrazolium bromide (MTT) approach. Transwell approach was used to measure cell invasion, and angiogenesis assay was used to detect the effect on angiogenesis. Luciferase report assay (LRA) measured targeting binding between miR-140-5p and VEGF-A. Comparing to tumor adjacent tissues, larynx carcinoma cells showed significantly decreased miR-140-5p expression. Agomir up-regulated miR-140-5p expression and weakened proliferation and invasion potency, and inhibited angiogenesis. Antagomir down-regulated miR-140-5p and presented the opposite results. Finally, LRA confirmed direct binding between miR-140-5p and VEGF-A. MiR-140-5p can target VEGF-A in larynx carcinoma cell line to inhibit cell invasion and angiogenesis. MiR-140-5p thus may work as the direct molecular target of larynx carcinoma.

Phytochemical and in vitro cytotoxicity analyses of wild bean (Glycine soja) ethanol extract using laryngeal cancer Hep-2 cells

Laryngeal cancer accounts for 0.6% of all cancer-related deaths worldwide. Wild soybean contains natural ingredients that have anti-cancer properties. This study aimed to determine the cytotoxicity of wild bean ethanol extract in inhibiting the growth of epidermal laryngeal cancer (Hep-2) cells for use as adjuvant therapy. Phytochemical analysis and thin-layer chromatography of the wild bean ethanol extract were conducted to identify the compounds contained in the extract. Wild bean ethanol extracts were tested against Hep-2 cancer cells in vitro. Cisplatin was used as a positive control. After incubation for 24 h, the inhibitory activities of the extracts were assessed with the MTT assay, and absorbances were determined with a microplate reader. Based on the phytochemical test results, the wild bean ethanol extract contains alkaloids, flavonoids, tannins, triterpenoids, saponins, and glycosides. The MTT assay showed that the median inhibitory concentration (IC50) of the wild bean ethanol extract was 118.061 μg/mL, and there were significant differences (p < 0.05) in the inhibitory activity at 800 μg/mL in comparison to the other treatment groups and controls. The IC50 value of cisplatin was 78.569 μg/mL. The wild bean ethanol extract had moderate cytotoxicity against Hep-2 laryngeal cancer cells and was less toxic than cisplatin.

Overexpressed miR-128a inhibits the proliferation of laryngeal cancer cells

Background: To identify the expression of miR-128a in laryngeal tumor tissues and to clarify the function of this miR on the proliferation capability of laryngeal cancer cells.

MicroRNA-1469, a p53-responsive microRNA promotes Genistein induced apoptosis by targeting Mcl1 in human laryngeal cancer cells

Genistein, a plant isoflavone, is reported to have therapeutic potentials in multiple cancers, However, the molecular mechanism underlying promoting cell apoptosis in laryngeal cancer remains unclear. In this study, we report that miR-1469 was induced by genistein in laryngeal cancer. Elevated miR-1469 promoted cell apoptosis and inhibited Mcl1 expression. In addition, we also observed that tumor suppressor p53 was increased under genistein treatment. Elevation of p53 promoted miR-1469 expression, leading to miR-1469 increase and Mcl1 decrease. Therefore, our findings suggest that genistein can suppress laryngeal cancer cell survival through p53 –miR-1469-Mcl1pathway.

Effect of miR-363 on the proliferation, invasion and apoptosis of laryngeal cancer by targeting Mcl-1.

To investigate the potential effect of miR-363 on the development of laryngeal cancer and to reveal the relevant mechanism. The expression level of miR-363 was detected in laryngeal cancer tissues and cells (TU-177), respectively. Luciferase assay was performed to evaluate the interaction between miR-363 and myeloid cell leukemia-1 (Mcl-1). The effect of the miR-363/Mcl-1 axis on TU-177 cells was determined by subsequent experiments including cell proliferation, invasion, apoptosis and the expression level of Mcl-1. In the present study, we found that miR-363 was both repressed in laryngeal cancer tissues and cells (TU-177). To find the regulating target of miR-363, we searched three publicly available algorithms, including TargetScan, miRDB, and microRNA. Results showed that Mcl-1 was a direct target of miR-363, and the Luciferase assay confirmed our suggestion. Subsequent experiments indicated that the decreased expression of Mcl-1 resulting from the up-regulation of miR-363 could deaccelerate cell proliferation and invasion, and accelerate cell apoptosis in laryngeal cancer cells. Our research revealed the suppressed function of miR-363 in laryngeal cancer by targeting Mcl-1. Meanwhile, we found that the restoration of miR-363 could serve as a potential therapeutic strategy for the treatment of laryngeal cancer.

Overexpression of Beclin1 inhibits proliferation and promotes apoptosis of human laryngeal squamous carcinoma cell Hep-2.

ObjectiveBeclin1 was previously found to be downregulated in human laryngeal cancer (LC) tissues, and it results in poor prognosis. This study aimed to further confirm the antitumor effects of Beclin1 in LC cell line Hep-2.Materials and methodsBeclin 1 was overexpressed in Hep-2 cells using liposomal transfection and confirmed using reverse transcription polymerase chain reaction and Western blotting. Then, cell proliferation and apoptosis were determined in control (untransfected), empty vector transfected, and Beclin1 overexpressed groups using MTT and flow cytometry procedure, respectively.ResultsThe expression of the Beclin1 gene in Hep-2 cells was significantly increased after vector transfection compared with control (1.173±0.046 vs 0.453±0.016, P<0.01) and empty vector (1.173±0.046 vs 0.440±0.021, P<0.01). Overexpression of Beclin1 inhibited proliferation at 4 days (0.619±0.051 vs 0.891±0.081 and 0.619±0.051 vs 0.878±0.105, P<0.01), 5 days (0.684±0.078 vs 1.127±0.094 and 0.684±0.078 vs 1.162±0.117, P<0.01), and 6 days (0.725±0.069 vs 1.168±0.103 and 0.725±0.069 vs 1.194±0.097, P<0.01) and promoted apoptosis (14.48%±1.42% vs 4.07%±0.66% and 14.48%±1.42% vs 4.39%±0.80%, P<0.01) in Hep-2 cells in comparison with the control and empty vector groups, respectively.ConclusionBeclin1 may be an underlying target for the treatment of LC. This study has provided some experimental basis for the gene therapy of LC.

PAG1 promotes the inherent radioresistance of laryngeal cancer cells via activation of STAT3

Phosphoprotein associated with glycosphingolipid-enriched microdomains 1(PAG1) is a ubiquitous protein that is essential for the development and progression of various malignancies. A previous study in our laboratory confirmed that PAG1 plays an important role in modulating the inherent radioresistance of laryngeal cancer cells, but the underlying mechanisms are still poorly defined. In this study, we found that PAG1 was significantly increased in laryngeal cancer tissues compared to adjacent non-tumor tissues (P < 0.05). The expression of PAG1 was positively correlated with lymph node metastasis (P < 0.05) and TNM stage (P < 0.05). High expression of PAG1 also predicted a poor prognosis in patients with laryngeal cancer. Moreover, gain-of-function and loss-of-function studies showed that PAG1 overexpression was able to promote growth, increase migration and invasion, and enhance inherent radioresistance of laryngeal cancer cells. Mechanistic investigations revealed that the activation of STAT3 was required for PAG1-mediated inherent radioresistance of laryngeal cancer. Inhibition of STAT3 activity with a chemical inhibitor sensitized radioresistant cells to radiation. Importantly, PAG1-integrin β1 complex was involved in the regulation of STAT3 activation. In addition, downregulation of PAG1 could suppress tumor growth and reverse inherent radioresistance in the nude mouse xenograft model. Taken together, these results suggested that PAG1 conferred inherent radioresistance by activating STAT3, which provided a novel therapeutic strategy for laryngeal cancer.