Invasion Of Laryngeal Cancer Cells Research Articles

Fibrinogen like protein-1 knockdown suppresses the proliferation and metastasis of TU-686 cells and sensitizes laryngeal cancer to LAG-3 blockade.

ObjectiveTo detect the expression of fibrinogen like protein-1 (FGL-1) in laryngeal cancer and evaluate its effect on tumor proliferation, metastasis, and antitumor immunity.MethodsELISA and immunohistochemistry were performed to detect FGL-1 expression in laryngeal cancer. The effects of FGL-1 knockdown on the proliferation, cell cycle progression, apoptosis, migration, and invasion of laryngeal cancer cells were evaluated by the CCK-8, colony formation, flow cytometry, Transwell migration, and western blot assays. We detected changes in tumorigenesis and drug response in vivo following FGL-1 knockdown as well as the effects of anti-LAG3 immunotherapy. Immunohistochemistry was performed to determine CD8 and LAG-3 expression in mouse tumor tissues.ResultsFGL-1 was highly expressed in the plasma and tumor tissues of laryngeal cancer patients. FGL-1 knockdown suppressed the proliferation of TU-686 cells and inhibited the migration and invasion of laryngeal cancer by blocking epithelial-to-mesenchymal transition. Moreover, silencing FGL-1 inhibited tumorigenicity in vivo and synergized with anti-LAG3 immunotherapy.ConclusionsWe confirmed the high expression of FGL-1 in laryngeal cancer and identified FGL-1 as a potential marker for immunotherapy in laryngeal cancer.

Effects of MiR-194-5p on the proliferation and invasion of laryngeal cancer cells by regulating the expression of smurf1 and activating the mTOR signaling pathway.

Laryngeal cancer has become the focus of research because of its high incidence rate and mortality rate. However, the research on miR-194-5p in laryngeal cancer is quite rare. The purpose of this study is to explore the effect of miR-194-5p on the proliferation and invasion of laryngeal cancer cells in order to find an effective way to treat laryngeal cancer. The results showed that miR-194-5p could activate the mTOR signaling pathway by regulating the expression of Smurf1, which had an effect on laryngeal cancer cells. The results showed that the absorbance of the miR-194-5p group was 0.38 lower than that of the NC group, which indicated that up-regulation of mir-194-5p could weaken the proliferation of laryngeal cancer cells. In addition, the average number of laryngeal cancer cells in NC and the miR-194-5p groups was 125.2 and 53.8, respectively, which indicated that miR-194-5p could reduce the number of laryngeal cancer cells passing through the basement membrane and their invasion ability.

Circular RNA circ-ABCB10 Promotes Proliferation and Inhibits Apoptosis of Laryngeal Carcinoma by Inhibiting KLF6.

Objective To explore the effect of circular RNA circ-ABCB10 on the proliferation and apoptosis of laryngeal carcinoma via inhibiting KLF6. Methods RT-qPCR assay was adopted to detect the expression of circ-ABCB10 and KFL6 in laryngeal carcinoma tissues and cell lines. Cell counting kit-8 (CCK-8) and clone formation assay were employed to detect laryngeal cancer cell viability and proliferation when circ-ABCB10 was silenced or upregulated. In this study, the apoptosis rate was detected by flow cytometry and the protein expression was detected by Western blotting. Wound healing and cross-hole invasion were used to study the migration and invasion of laryngeal cancer cells when circ-ABCB10 was silenced or upregulated. Results The results of RT-qPCR detection indicated that the expression of circ-ABCB10 in all three laryngeal carcinoma cells was downregulated by 3.2 times compared with that of HaCat cells. There is low expression of circ-ABCB10 in most laryngeal carcinoma tissues, the diagnostic cutoff value of circ-ABCB10 is 0.0008, the area under the curve is 0.718, the sensitivity is 0.981, and the specificity is 0.556. The expression level of KLF6 in laryngeal carcinoma is on the rise, which is significantly higher compared to healthy tissues (P < 0.05); 48 hours after transfection, RT-qPCR analysis confirmed the transfection efficiency, and upregulation of circ-ABCB10 could significantly promote cell proliferation. Compared with the control group, silencing circ-MTCL1 could inhibit cell proliferation, overexpression of circ-ABCB10 promoted cell migration, and downregulation of circ-ABCB10 significantly inhibited cell movement (P < 0.001). Upregulation of circ-ABCB10 significantly enhanced the invasiveness and motility of laryngeal cancer cells, while downregulation of circ-ABCB10 was the opposite. Compared with the KLF6 NC group, KLF6 level increased significantly in the KLF6 group, while cell viability, colony formation, scratch healing rate, invasive cell number, and Bcl-2 expression level decreased significantly in the KLF6 group, while apoptosis rate and Bax expression level increased significantly (P < 0.05). KLF6 level in the si-circ-ABCB10+anti-KLF6 group was significantly lower than that in the si-circ-ABCB10+anti-KLF6-NC group (P < 0.05). Meanwhile, the cell activity, colony formation number, cell scratch healing rate, number of invaded cells, and Bcl-2 all indicated an upward trend, while the cell apoptosis rate and Bax expression indicated a downward trend (P < 0.05). Conclusion The expression of circ-ABCB10 in laryngeal carcinoma was significantly higher compared to that in paracancerous tissues. Silencing circ-ABCB10 could significantly inhibit the growth and proliferation of laryngeal adenocarcinoma cells, while overexpression of circ-ABCB10 could significantly promote the growth of laryngeal adenocarcinoma cells, probably by inhibiting KLF6 to enhance the proliferation of laryngeal carcinoma and inhibit apoptosis.

AQP9 and ZAP70 as immune-related prognostic biomarkers suppress proliferation, migration and invasion of laryngeal cancer cells

BackgroundLaryngeal cancer represents a common malignancy that originates from the larynx, with unfavorable prognosis. Herein, this study systematically analyzed the immune signatures of laryngeal cancer and to evaluate their roles on tumor progression.MethodsDifferentially expressed immune-related genes (IRGs) were screened between laryngeal cancer and normal tissues from TCGA dataset. Then, two prognosis-related IRGs AQP9 and ZAP70 were analyzed by a series of survival analysis. Based on them, molecular subtypes were constructed by unsupervised cluster analysis. Differences in survival outcomes, HLA expression and immune cell infiltrations were assessed between subtypes. Expression of AQP9 and ZAP70 was validated in laryngeal cancer tissues and cells by RT-qPCR and immunohistochemistry. After silencing and overexpressing AQP9 and ZAP70, CCK-8, EdU, wound healing and transwell assays were performed in TU212 and LCC cells.ResultsTotally, 315 IRGs were abnormally expressed in laryngeal cancer. Among them, AQP9 and ZAP70 were distinctly correlated to patients’ prognosis. Two subtypes were developed with distinct survival outcomes, HLA expression and immune microenvironment. Low expression of AQP9 and ZAP70 was confirmed in laryngeal cancer. AQP9 and ZAP70 up-regulation distinctly suppressed proliferation, migration, and invasion of laryngeal cancer cells. The opposite results were investigated when their knockdown.ConclusionsOur findings revealed the roles of AQP9 and ZAP70 in progression of laryngeal cancer, and suggested that AQP9 and ZAP70 could potentially act as candidate immunotherapeutic targets for laryngeal cancer.

MicroRNA-194-5p on Proliferation and Invasion of Laryngeal Cancer Cells by Regulating the Expression of SMURF1 and Activating the mTOR Signalling Pathway

Laryngeal cancer has become the focus of research because of its high incidence rate and mortality rate. However, the research on microRNA-194-5p in laryngeal cancer is quite rare. The purpose of this study is to explore the effect of microRNA-194-5p on the proliferation and invasion of laryngeal cancer cells, in order to find an effective way to treat laryngeal cancer. The results showed that microRNA-194-5p could activate mammalian target of rapamycin signaling pathway by regulating the expression of smad ubiquitin regulatory factor 1, which had an effect on laryngeal cancer cells. The results showed that the absorbance of microRNA-194-5p group was 0.38 lower than that of negative control group, which indicated that up regulation of microRNA-194-5p could weaken the proliferation of laryngeal cancer cells. In addition, the average number of laryngeal cancer cells in negative control group and microRNA-194-5p group was 125.2 and 53.8 respectively, which indicated that microRNA-194-5p could reduce the number of laryngeal cancer cells passing through basement membrane and their invasion ability.

Development and Validation of an Immune-Related Signature for the Prediction of Recurrence Risk of Patients With Laryngeal Cancer.

ObjectiveOur purpose was to develop and verify an immune-related signature for predicting recurrence risk of patients with laryngeal cancer.MethodsRNA-seq data of 51 recurrence and 81 non-recurrence laryngeal cancer samples were downloaded from TCGA database, as the training set. Microarray data of 34 recurrence and 75 non-recurrence cancer samples were obtained from GEO dataset, as the validation set. Single factor cox regression was utilized to screen prognosis-related immune genes. After LASSO regression analysis, an immune-related signature was constructed. Recurrence free survival (RFS) between high- and low- recurrence risk patients was presented, followed by ROC. We also evaluated the correlation between immune infiltration and the signature using the CIBERSORT algorithm. The genes in the signature were validated in laryngeal cancer tissues by western blot or RT-qPCR. After RCN1 knockdown, migration and invasion of laryngeal cancer cells were investigated.ResultsTotally, 43 prognosis-related immune genes were identified for laryngeal cancer. Among them, eight genes were used for constructing a prognostic signature. High risk group exhibited a higher recurrence risk than low risk group. The AUC for 1-year was separately 0.803 and 0.715 in the training and verification sets, suggesting its well efficacy for predicting the recurrence. Furthermore, this signature was closely related to distinct immune cell infiltration. RCN1, DNAJA2, LASP1 and IBSP were up-regulated in laryngeal cancer. RCN1 knockdown restrained migrated and invasive abilities of laryngeal cancer cells.ConclusionOur findings identify a reliable immune-related signature that can predict the recurrence risk of patients with laryngeal cancer.

Long noncoding RNA NEAT1 inhibits the acetylation of PTEN through the miR-524-5p /HDAC1 axis to promote the proliferation and invasion of laryngeal cancer cells.

Long noncoding RNA nuclear paraspeckle assembly transcript 1 (lncRNA NEAT1) is abnormally expressed in numerous tumors and functions as an oncogene, but the role of NEAT1 in laryngocarcinoma is largely unknown. Our study validated that NEAT1 expression was markedly upregulated in laryngocarcinoma tissues and cells. Downregulation of NEAT1 dramatically suppressed cell proliferation and invasion through inhibiting miR-524-5p expression. Additionally, NEAT1 overexpression promoted cell growth and metastasis, while overexpression of miR-524-5p could reverse the effect. NEAT1 increased the expression of histone deacetylase 1 gene (HDAC1) via sponging miR-524-5p. Mechanistically, overexpression of HDAC1 recovered the cancer-inhibiting effects of miR-524-5p mimic or NEAT1 silence by deacetylation of tensin homolog deleted on chromosome ten (PTEN) and inhibiting AKT signal pathway. Moreover, in vivo experiments indicated that silence of NEAT1 signally suppressed tumor growth. Taken together, knockdown of NEAT1 suppressed laryngocarcinoma cell growth and metastasis by miR-524-5p/HDAC1/PTEN/AKT signal pathway, which provided a potential therapeutic target for laryngocarcinoma.

Hsa-circ_0058106 induces EMT and metastasis in laryngeal cancer via sponging miR-153 and inducing Twist1 nuclear translocation.

A new circular RNA (hsa-circ_0058106) has been found to be upregulated in laryngeal cancer, but its function remains unknown. Here, we explored its role in the metastasis of laryngeal cancer. The level of hsa-circ_0058106 in laryngeal cancer was detected by qRT-PCR. The effect of hsa-circ_0058106 silencing on the metastasis of laryngeal cancer was assessed using scratch wound healing and transwell assays, as well as nude mouse lung metastasis models. Fluorescence in situ hybridization was employed to analyze the cellular localization of hsa-circ_0058106. A luciferase activity assay was used to assess binding between hsa-circ_0058106 and miR-153. Interaction between hsa-circ_0058106 and Twist1 was confirmed using RNA pull-down and RNA immunoprecipitation assays. A high level of hsa-circ_0058106 was found to be associated with lymph node metastasis and advanced clinical stages. Stable knockdown of hsa-circ_0058106 inhibited the migration and invasion of laryngeal cancer cells in vitro and in vivo. Moreover, we found that downregulation of hsa-circ_0058106 suppressed epithelial-mesenchymal transition (EMT), which was underscored by the observation that hsa-circ_0058106 silencing led to decreases in the expression of N-cadherin and Vimentin, and an increase in the expression of E-cadherin. Mechanistically, we found that hsa-circ_0058106 can specifically bind to miR-153 and regulate Snail1 expression by acting as a miR-153 sponge. In addition, we found that knockdown of hsa-circ_0058106 blocked the nuclear translocation of Twist1. Our results indicate that hsa-circ_0058106 induces EMT and metastasis in laryngeal cancer by sponging miR-153 and inducing Twist1 nuclear translocation. These observations provide new insights into the regulatory effects of circRNAs in laryngeal cancer metastasis.

Magnesium Isoglycyrrhizinate Induces an Inhibitory Effect on Progression and Epithelial-Mesenchymal Transition of Laryngeal Cancer via the NF-κB/Twist Signaling.

BackgroundMagnesium isoglycyrrhizinate (MI) was extracted from roots of the plant Glycyrrhiza glabra, which displays multiple pharmacological activities such as anti-inflammation, anti-apoptosis, and anti-tumor. Here, we aimed to investigate the effect of MI on the progression and epithelial–mesenchymal transition (EMT) of laryngeal cancer.MethodsForty laryngeal cancer clinical samples were used. The role of MI in the proliferation of laryngeal cancer cells was assessed by MTT assay, Edu assay and colony formation assay. The function of MI in the migration and invasion of laryngeal cancer cells was tested by transwell assays. The effect of MI on apoptosis of laryngeal cancer cells was determined by cell apoptosis assay. The impact of MI on tumor growth in vivo was analyzed by tumorigenicity analysis using Balb/c nude mice. qPCR and Western blot analysis were performed to measure the expression levels of gene and protein, respectively.ResultsWe identified that EMT-related transcription factor Twist was significantly elevated in the laryngeal cancer tissues. The expression of Twist was also enhanced in the human laryngeal carcinoma HEP-2 cells compared with that in the primary laryngeal epithelial cells. The high expression of Twist was remarkably correlated with poor overall survival of patients with laryngeal cancer. Meanwhile, our data revealed that MI reduced cell proliferation, migration and invasion and enhanced apoptosis of laryngeal cancer cells in vitro. Moreover, MI decreased transcriptional activation and the expression levels of NF-κB and Twist, and alleviated EMT in vitro and in vivo. MI remarkably inhibited tumor growth and EMT of laryngeal cancer cells in vivo.ConclusionMI restrains the progression of laryngeal cancer and induces an inhibitory effect on EMT in laryngeal cancer by modulating the NF-κB/Twist signaling. Our finding provides new insights into the mechanism by which MI inhibits laryngeal carcinoma development, enriching the understanding of the anti-tumor function of MI.

Quinolinol-platinum (II) complex suppresses survival and invasion of laryngeal cancer cells via targeting YESassociated protein expression

Purpose: To investigate the anti-proliferative potential of quinolinol-platinum (II) complex {QN-Pt (II)} in laryngeal cancer cells.Methods: The inhibitory potential of QN-Pt (II) on the proliferation of laryngeal cancer cells was determined using 2H-tetrazolium, 2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl bromide (MTT) and colony formation assays. Inhibition of cell migration was determined using wound-healing assay, while changes in LC3-II and p62 expressions were assessed by western blotting.Results: QN-Pt (II) inhibited the viability of TU212 and M4e laryngeal cancer cells in the concentration range of 2 to 10 μM (p &lt; 0.05). Moreover, it suppressed the colony formation potential and migration of TU212 and M4e cells. The QN-Pt (II) treatment increased the proportion of TU212 and M4e cells in G1/G0 phase of the cell cycle, but decreased the cell proportion in S and G2/M phase (p &lt; 0.05). Treatment with 10 μM QN-Pt (II) increased the expression of LC3-II, but downregulated P62 expression in TU212 and M4e cells. The expression of Yes-associated protein was inhibited in TU212 and M4e cells on treatment with QN-Pt (II). However, transfection of YAP-cDNA into TU212 and M4e cells reversed the inhibitory effect of QN-Pt (II) on cell proliferation (p &lt; 0.05). Moreover, YAP-cDNA transfection suppressed LC3II expression, inhibited YAP phosphorylation and promoted P62 expression in QN-Pt (II)-treated TU212 and M4e cells (p &lt; 0.05).Conclusion: QN-Pt (II) suppresses laryngeal cancer cell viability via arrest of cell cycle and activation of apoptosis. Moreover, QN-Pt (II) targets Yes-associated protein in laryngeal cancer cells. Thus, QN-Pt (II) is a potential therapeutic agent for laryngeal cancer.&#x0D; Keywords: Laryngeal cancer, YAP-protein, Apoptosis, Phosphorylation, Therapeutic agent

MicroRNA-107 inhibits the proliferation and invasion of laryngeal squamous cell carcinoma cells by targeting CACNA2D1

Objective:To detect the expression of microRNA-107 （miR-107） and the calcium channel protein gene CACNA2D1 in laryngeal cancer tissues, to investigate the targeting relationship between miR-107 and CACNA2D1, and to analyze the effects of miR-107 on the proliferation, invasion and colony forming ability of laryngeal cancer cells. Method:Laryngeal cancer tissues and normal adjacent tissue samples from 40 patients with laryngeal cancer were collected, and qRT-PCR was used to detect the expression of miR-107 and CACNA2D1; Western Blot assay to detect the expression of α2δ1 in the above two tissues; the dual-luciferase reporter gene was used to detect the regulatory effect of miR-107 on CACNA2D1; after overexpression or knockdown of miR-107 in human laryngeal cancer cells TU212 and TU686, changes in the proliferation, clone formation, and invasion ability of laryngeal cancer cells were detected. Result:The expression of miR-107 in laryngeal cancer tissues was significantly lower than that in adjacent normal tissues, while the expression of CACNA2D1 was just the opposite, the difference was statistically significant （P<0.05）; the expression of α2δ1 in laryngeal cancer tissues is significantly higher than in normal tissues（P<0.05）; dual-luciferase reporter experiments confirmed that miR-107 binds to the 3'-UTR （202-209, 902-908） of the CACNA2D1 mRNA, thereby inhibiting the expression of CACNA2D1 and its biological effects; cell experiments showed that the proliferation, invasion, and clone formation of laryngeal cancer cells were significantly reduced after miR-107 overexpression （P<0.05）, and the cell proliferation, clone formation, and invasion were significantly enhanced after miR-107 was knocked down （P<0.05） . Conclusion:miR-107 inhibits the proliferation, clone formation, and invasion of laryngeal cancer cells by targeting CACNA2D1.

Targeted-knockdown of Yes-associated protein inhibits the Warburg effect and the invasion of laryngeal cancer cells

Objective: To investigate the migration and invasion behaviors of Hep-2 after the targeted knockdown of yes-associated protein (YAP). Methods: Hep-2 cells were knock-downed for YAP by shRNA as YAP-shRNA group, Hep-2 treated with non-specific shRNA as YAP-NC group, and Hep-2 with no treatment as control. Glucose uptake and lactate production in the cells were examined to assess Warburg effect. The migration and invasion behaviors of cells in three groups were observed. The expressions of vimentin and E-cadherin were detected by RT-PCR and Western Blot. The statistical software GraphPad Prism 7.0 was used to analyze significance of data. Two tailed Student' s t-tests was used to determine significance when only two groups were compared. P values of less than 0.05 was considered statistically significant. Results: Downregulation of YAP led to a obvious decrease in glucose uptake [(18.51±1.72)%] and lactate production [103.40±8.32] in Hep-2 cells compared with control [(41.20±1.11)% and 743.69±19.49, t=19.20 and 52.33, respectively, both P<0.01] and YAP-NC group [(39.60±0.78)% and 705.22±17.20, t=19.34 and 54.56, respectively, both P<0.01]. Compared with the control group (78.32±4.04) and YAP-NC group (77.28±3.11), the scratch healing ability of Hep-2 cells was significantly decreased in YAP-shRNA group (44.71±4.68). The P value was less than 0.01 (t=9.42 and 10.04). The number of cells with YAP-shRNA (33.30±4.19) passing through compartments was remarkable fewer than the control group (133.71±6.72) and YAP-NC group (126.32±4.21). The P value was less than 0.01 (t=21.96 and 27.13). The expression of E-cadherin protein in cells of YAP-shRNA group (6.16±0.11) was up-regulated compared with control (0.97±0.10, t=35.70, P<0.01) and YAP-NC group (1.13±0.09, t=36.28, P<0.01), while the expression of vimentin protein in cells of YAP-shRNA group (1.08±0.09) was down-regulated compared with control (5.67±0.12, t=29.91, P<0.01) and YAP-NC group (5.51±0.12, t=29.04, P<0.01). Conclusions: The down-regulation of YAP in Hep-2 inhibits the migration and invasion of cells via suppressing Warburg and EMT program.

Small Nucleolar RNA Host Gene 12 (SNHG12) Promotes Proliferation and Invasion of Laryngeal Cancer Cells via Sponging miR-129-5p and Potentiating WW Domain-Containing E3 Ubiquitin Protein Ligase 1 (WWP1) Expression.

BackgroundThe clinical significance and biological function of long noncoding RNA SNHG12 have not been identified in laryngeal squamous cell carcinoma (LSCC).Material/MethodsExpression levels of SNHG12, miR-129-5p, and WWP1 in LSCC tissues or cells were tested by RT-qPCR. MTT assay, flow cytometry, and Transwell assay were used to identify the progression of LSCC cells in vitro. Luciferase reporter assay was used to assess the associations among SNHG12, WWP1, and miR-129-5p.ResultsSNHG12 was significantly overexpressed in LSCC tissues compared with adjacent normal tissues. The expression level of SNHG12 was significantly associated with T classification, lymph node metastasis, and cancer stage of LSCC. High expression of SNHG12 predicted shorter disease-free survival. Suppressing SNHG12 using siRNA inhibited proliferation and invasion and promoted apoptosis in the AMC-HN-8 LSCC cell line. SNHG12, mainly located in cytoplasm of AMC-HN-8 cells, was validated by dual luciferase reporter test and RT-qPCR to directly interact with miR-129-5p. Inhibition of miR-129-5p significantly increased proliferation and invasion of AMC-HN-8 cells and ameliorated the suppressive effects of si-SNHG12. Luciferase assay showed that miR-129-5p was able to combine with the 3′UTR region of WWP1, which is generally regarded as an E3 ubiquitin protein ligase. RT-qPCR and Western blot showed that WWP1 was positively regulated by SNHG12 and negatively regulated by miR-129-5p at the mRNA level and protein level. Overexpression of WWP1 significantly increased proliferation and invasion of laryngeal cancer cells. Moreover, when SNHG12 was suppressed, rescue of WWP1 restored the proliferation and invasion abilities of AMC-HN-8 cells.ConclusionsOur study demonstrated that SNHG12 promoted LSCC cells progression via sponging miR-129-5p and potentiating WWP1 expression.

Knockdown of hsa_circ_0023028 inhibits cell proliferation, migration, and invasion in laryngeal cancer by sponging miR-194-5p.

Emerging evidences have proposed that circular RNAs (circRNAs) play a major role in carcinogenesis. Hsa_circ_0023028 has been reported to be aberrantly expressed in laryngeal cancer (LCa). However, the role and the mechanism of hsa_circ_0023028 in LCa have not been adequately studied. In the present study, we demonstrated that hsa_circ_0023028 expression was up-regulated in LCa tissues and cell lines. miR-194-5p was down-regulated in LCa cells. Functionally, knockdown of hsa_circ_0023028 inhibited the proliferation, migration, and invasion of LCa cells, as evidenced by the reduced number of 5-Ethynyl-2′-deoxyuridine (EdU)-positive cells and decreased number of migrated and invaded cells. Additionally, hsa_circ_0023028 was identified as an miR-194-5p sink. A negative correlation between miR-194-5p and hsa_circ_0023028 expression was observed in LCa tissues. Besides, down-regulation of miR-194-5p attenuated the inhibitory effects of hsa_circ_0023028 silencing on LCa cell proliferation, migration, and invasion. In summary, hsa_circ_0023028 functions as an miR-194-5p sponge to promote the proliferation, migration, and invasion of LCa cells.

Long non-coding RNA Dleu2 affects proliferation, migration and invasion ability of laryngeal carcinoma cells through triggering miR-16-1 pathway.

Laryngeal cancer is a common malignant tumor in the head and neck, which affects swallowing, breathing, and pronunciation function. In recent years, many long non-coding RNAs (lncRNAs) have been shown to be involved in the progression of cancer with the development of gene sequencing, transcriptomics, and bioinformatics. LncRNA Dleu2 is a cancer-related lncRNA that down-regulates tumor progression in a variety of cancers. However, its possible effects and related signaling pathway in the development of laryngeal cancer are not clear. Real-time PCR was applied to test lncRNA Dleu2 and microRNA-16-1 (miR-16-1) expressions in laryngeal carcinoma tissues. LncRNA Dleu2 and miR-16-1 levels were over-expressed by transfection. Cell proliferation was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Cell migration was evaluated by using wound-healing assay. Cell invasion was determined by using transwell assay. LncRNA Dleu2 and miR-16-1 levels were significantly declined in the laryngeal carcinoma tissue compared to para-carcinoma tissue (p < 0.05). LncRNA Dleu2 and miR-16-1 up-regulation significantly reduced cell proliferation, migration, and invasion compared to the control group (p < 0.05). Ago-miR16-1 transfection significantly enhanced the luciferase activity of wild-type Dleu2 compared to control group (p < 0.05), suggesting their interaction with each other. LncRNA Dleu2 influences the proliferation, migration, and invasion of laryngeal cancer cells through miR-16-1. Therefore, lncRNA Dleu2 and miR-16-1 may serve as potential biomarkers and targets for laryngeal cancer diagnosis and treatment.

NOB1 silencing inhibits the growth and metastasis of laryngeal cancer cells through the regulation of JNK signaling pathway.

Nin one binding protein (NOB1) plays important roles in the synthesis and degradation of proteins, thus having effects on the cellular process. In the present study, the expression level of NOB1 in laryngeal cancer patients was detected by quantitative PCR and western blotting, and the effect of NOB1 on growth and metastasis of laryngeal cancer cells was explored. Silence of NOB1 was found to inhibit the proliferation of laryngeal cancer cells, arrest cell cycle and induce cell apoptosis. NOB1 silence was also found to inhibit the migration and invasion of laryngeal cancer cells and to downregulate the protein levels of matrix metalloproteinases (MMPs)-2 and MMP-9. Further mechanism study revealed that the JNK signaling pathway was involved in the function of NOB1. Our present results suggest that NOB1 plays an oncogenic role in laryngeal cancer cells through the regulation of JNK signaling pathway, and lays a theoretical foundation for further exploration of NOB1.

Influence of chitosan nanoparticle-mediated C-erbB-2 gene silencing on invasion and apoptosis of Hep-2 cells

We aimed to measure the invasion ability of Hep-2 laryngeal cancer cells after treatment with C-erbB-2-small interfering RNA (siRNA)-chitosan nanoparticles, and assess the applied value of chitosan nanoparticle-mediated C-erbB-2 interference in inhibiting laryngeal cancer invasion and metastasis. Nanoparticles of approximately 100 nm, comprising C-erbB-2 siRNA packaged with chitosan, were prepared and used to treat Hep-2 cells. Silencing of C-erbB-2 was detected by western blot and polymerase chain reaction. Cell invasion and apoptosis were estimated by transwell assay and flow cytometry, respectively. C-erbB-2-siRNA-chitosan nanoparticles significantly down-regulated C-erbB-2 expression in Hep-2 cells (P < 0.05), and cell invasion was noticeably decreased. Moreover, they significantly induced apoptosis of the Hep-2 cells (P < 0.05). Chitosan nanoparticle-mediated C-erbB-2 gene interference can inhibit the invasion of laryngeal cancer cells and induce their apoptosis.

SOX2 promotes the migration and invasion of laryngeal cancer cells by induction of MMP-2 via the PI3K/Akt/mTOR pathway

SOX2 is a high mobility group box containing transcription factor that has been reported to be aberrantly overexpressed in various human malignancies, including laryngeal squamous cell carcinoma (LSCC). However, the potential role of SOX2 in LSCC migration and invasion remains to be elucidated. In the present study, we generated stable transformants of human LSCC cells constitutively overexpressing SOX2 and investigated the effects of SOX2 overexpression on migration and invasion in LSCC cells as well as the possible underlying mechanisms. We found that ectopic overexpression of SOX2 in LSCC cells enhanced their migratory and invasive ability invitro, accompanied by increased expression and activity of matrix metalloproteinase (MMP)-2. Meanwhile, SOX2-induced cell migration and invasion were significantly abrogated by a neutralizing anti-MMP-2 antibody or small interfering RNA targeting MMP-2. Furthermore, overexpression of SOX2 induced phosphorylation of Akt and mammalian target of rapamycin (mTOR), which are downstream effectors of the PI3K pathway. Finally, LY294002, an inhibitor of PI3K, also markedly abolished SOX2-induced activation of the Akt/mTOR pathway and increased cell invasion and MMP-2 expression. Taken together, we conclude that SOX2 promotes migration and invasion of laryngeal cancer cells by inducing MMP-2 via the PI3K/Akt/mTOR pathway. Our findings suggest that SOX2 may serve as a potential therapeutic target for LSCC.

Inhibition of Laryngeal Cancer Cell Invasion and Growth with Lentiviral-Vector Delivered Short Hairpin RNA Targeting Human MMP-9 Gene

The purpose of this study was to explore the inhibiting role of MMP-9 gene silence in the invasive ability and growth of laryngeal squamous cell carcinoma (LSCC) by lentivirus mediated RNA interference. MMP-9-RNAi-lentivirus and the control lentivirus (GFP-lentivirus) were transfected into Hep-2 cells. Gelatin zymography showed the proteins expression of MMP-9 were knockdown in the MMP-9 siRNA transfected Hep-2 cells. The invasive activity and viability of MMP-9 siRNA treated Hep-2 cells were decreased than the control cells measured with modified Boyden chamber assay and MTT assay. In animal experiment, 20 nude mice bearing Hep-2 cell tumor were randomly separated into the experimental and the control groups. The former were intratumorally injected with MMP-9-RNAi-lentivirus, and the later were injected with equivalent dose of GFP-lentivirus. Results showed the average weight and volume of tumor in MMP-9-RNAi-lentivirus treated group were significantly lower than those in the control group (P <. 01). The protein expressions of MMP-9 were downregulated in tumors of MMP-9-RNAi-lentivirus treatment. The PCNA index was obviously lower in the tumors of treated group than that in the control group (P <. 01). These results suggest that MMP-9 gene silence by lentivirus mediated RNA interference can inhibit invasion and growth of LSCC.