Prognosis Of Laryngeal Squamous Cell Carcinoma Patients Research Articles

Characterization of Cancer Stem Cells in Laryngeal Squamous Cell Carcinoma by Single-cell RNA Sequencing.

Cancer stem cells (CSCs) constitute a pivotal element within the tumor microenvironment (TME), driving the initiation and progression of cancer. However, the identification of CSCs and their underlying molecular mechanisms in laryngeal squamous cell carcinoma (LSCC) remains a formidable challenge. Here, we employed single-cell RNA sequencing of matched primary tumor tissues, paracancerous tissues, and local lymph nodes from three LSCC patients to comprehensively characterize the CSCs in LSCC. Two distinct clusters of stem cells originating from epithelial populations were delineated and verified as CSCs and normal stem cells (NSCs), respectively. CSCs were abundant in the paracancerous tissues compared to those in the tumor tissues. CSCs showed high expression of stem cell marker genes such as PROM1, ALDH1A1, and SOX4, and increased the activity of tumor-related hypoxia, Wnt/β-catenin, and Notch signaling pathways. We then explored the intricate crosstalk between CSCs and the TME cells and identified targets within the TME that related with CSCs. We also found eight marker genes of CSCs that were correlated significantly with the prognosis of LSCC patients. Furthermore, bioinformatics analyses showed that drugs such as erlotinib, OSI-027, and ibrutinib selectively targeted the CSC-specifically expressed genes. In conclusion, our results represent the first comprehensive characterization of CSC properties in LSCC at the single-cell level.

M6A/HOXA10-AS/ITGA6 axis aggravates oxidative resistance and malignant progression of laryngeal squamous cell carcinoma through regulating Notch and Keap1/Nrf2 pathways

As the second most prevalent malignant tumor of head and neck, laryngeal squamous cell carcinoma (LSCC) imposes a substantial health burden on patients worldwide. Within recent years, resistance to oxidative stress and N6-methyladenosine (m6A) of RNA have been proved to be significantly involved in tumorigenesis. In current study, we investigated the oncogenic role of m6A modified long non coding RNAs (lncRNAs), specifically HOXA10-AS, and its downstream signaling pathway in the regulation of oxidative resistance in LSCC. Bioinformatics analysis revealed that heightened expression of HOXA10-AS was associated with the poor prognosis in LSCC patients, and N (6)-Methyladenosine (m6A) methyltransferase-like 3 (METTL3) was identified as a factor in promoting m6A modification of HOXA10-AS and further intensify its RNA stability. Mechanistically, HOXA10-AS was found to play as a competitive endogenous RNA (ceRNA) by sequestering miR-29 b-3p and preventing its downregulation of Integrin subunit alpha 6 (ITGA6), ultimately enhancing the oxidative resistance of tumor cells and promoting the malignant progression of LSCC. Furthermore, our research elucidated the mechanism by which ITGA6 accelerates Keap1 proteasomal degradation via enhancing TRIM25 expression, leading to increased Nrf2 stability and exacerbating its aberrant activation. Additionally, we demonstrated that ITGA6 enhances γ-secretase-mediated Notch signaling activation, ultimately promoting RBPJ-induced TRIM25 transcription. The current study provides the evidence supporting the effect of m6A modified HOXA10-AS and its downstream miR-29 b-3p/ITGA6 axis on regulating oxidative resistance and malignant progression in LSCC through the Notch and Keap1/Nrf2 pathways, and proposed that targeting this axis holds promise as a potential therapeutic approach for treating LSCC.

Development and validation of a model to predict the risk of recurrence in patients with laryngeal squamous cell carcinoma after total laryngectomy.

Recurrence is still the main obstacle to the survival of laryngeal squamous cell carcinoma (LSCC) patients who have undergone a total laryngectomy. Previous models for recurrence prediction in patients with LSCC were based on pathological information, while the role of easily accessible inflammatory markers in the prognosis of LSCC patients has rarely been reported. This study sought to develop and validate a model to predict the risk of recurrence in LSCC patients who underwent total laryngectomy. A total of 204 LSCC patients who underwent a total laryngectomy were included in this retrospective cohort study. Demographics, pathology, and inflammatory markers of patients were collected. All the patients were randomly divided into a training set and a test set at a ratio of 4:1. Patients were followed up for 3 years after surgery or until death occurred during this period. The random-forest method was used to develop a predictive model. The performance of the model was evaluated by calculating the area under the receiver operating characteristic (ROC) curve (AUC) with the 95% confidence interval (CI), sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Of the 204 LSCC patients, 56 (27.45%) patients had a recurrence. The random-forest prediction model was an all-factor model, and the most important predictors of the model were the albumin/globulin ratio (AGR), neutrophil/lymphocyte ratio (NLR), and platelet/lymphocyte ratio (PLR), with proportions of 0.121, 0.100, and 0.092, respectively. The AUCs of the model in predicting the recurrence of LSCC in the training set and the test set were 0.960 (95% CI, 0.931-0.989) and 0.721 (95% CI, 0.716-0.726), respectively. The sensitivity, specificity, accuracy, PPV, and NPV of the model in the test set were 0.750 (95% CI, 0.505-0.995), 0.690 (95% CI, 0.521-0.858), 0.707 (95% CI, 0.568-0.847), 0.500 (95% CI, 0.269-0.921), and 0.870 (95% CI, 0.732-1.000), respectively. A model to predict the risk of recurrence in LSCC patients who have undergone a total laryngectomy was established, and inflammatory markers AGR, NLR, and PLR play an important role in the predictive model.

Overexpression of B7-H3 Is Associated With Poor Prognosis in Laryngeal Cancer.

The immune checkpoint molecule, B7-H3, which belongs to the B7 family, has been shown to be overexpressed in various cancers. Its role in tumors is not well defined, and many studies suggest that it is associated with poor clinical outcomes. The effect of B7-H3 on laryngeal cancer has not been reported. This study investigated the expression of B7-H3 in laryngeal squamous cell carcinoma (LSCC), and its relationship with clinicopathological factors and prognosis of LSCC patients. The gene expression quantification data and clinical data of LSCC retrieved from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) database were analyzed to determine the diagnostic and prognostic roles of B7-H3. Quantitative real-time polymerase chain reaction (qRT-PCR) was then performed to determine the gene expression level of B7-H3 between LSCC tissues and paired normal adjacent tissues. In addition, TCGA RNA-seq data was analyzed to evaluate the expression level of B7 family genes. Next, the protein expression of B7-H3 and CD8 in LSCC was determined using immunohistochemistry and immunofluorescence. qRT-PCR results showed that the expression level of B7-H3 mRNA was significantly higher in LSCC tissues than in adjacent normal tissues. Similar results were obtained from the TCGA analysis. The expression of B7-H3 was significantly associated with T stage, lymph node metastasis, and pathological tumor node metastasis (TNM) stage, and it was also an independent factor influencing the overall survival time (OS) of patients with LSCC. In addition, B7-H3 was negatively correlated with CD8+T cells. These results show that B7-H3 is upregulated in LSCC. Therefore, B7-H3 may serve as a biomarker of poor prognosis and a promising therapeutic target in LSCC.

Circular RNA 103862 Promotes Proliferation and Invasion of Laryngeal Squamous Cell Carcinoma Cells Through the miR-493-5p/GOLM1 Axis.

Accumulating evidence suggests that circular RNAs (circRNAs) may be a key contributor to oncogenesis. Yet, the function of circRNAs in laryngeal squamous cell carcinoma (LSCC) is still not clear. In this study, we examined the function of circRNA_103862 in LSCC progression by analyzing the tissue specimens collected from a patient with LSCC by using different LSCC cell models in vitro and an LSCC xenograft model in nude mice. We found that circRNA_103862 was frequently upregulated in the tissues of LSCC and was correlated with metastasis and prognosis of LSCC patients. Furthermore, circRNA_103862 downregulation could reduce proliferation, migration, and invasion ability of LSCC cells. In terms of mechanism exploration, miR-493-5p was sponged by circRNA_103862. Rescue experiments also showed that circRNA_103862 could achieve a carcinogenic effect by regulating miR-493-5p. Moreover, a luciferase reporter analysis showed that Golgi membrane protein 1 (GOLM1) is a downstream effector of miR-493-5p. In conclusion, our data suggested that circRNA_103862 promotes the proliferation of LSCC through targeting the miR-493-5p/GOLM1 axis, and it might serve as a potential prognosis marker and therapy target for LSCC.

STYK1 promotes the malignant progression of laryngeal squamous cell carcinoma through targeting TGF-β1.

The aim of this study was to uncover the expression characteristic and biological function of STYK1 in the progression of laryngeal squamous cell carcinoma (LSCC), and to explore the underlying mechanism. Expression level of STYK1 in 44 paired LSCC and adjacent normal tissues was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The relationship between STYK1 level and clinical parameters of LSCC patients was analyzed. Subsequently, the regulatory effect of STYK1 on the proliferative ability of AMC-HN-8 and Hep-2 cells was evaluated by cell counting kit-8 (CCK-8) assay and colony formation assay. Dual-Luciferase reporter gene assay and rescue experiments were conducted to uncover the role of STYK1/TGF-β1 axis in regulating the progression of LSCC. STYK1 was significantly up-regulated in LSCC tissues than that of adjacent normal tissues (p<0.05). LSCC patients with high expression level of STYK1 exhibited significantly higher clinical stage and lower survival rate (p<0.05). Knockdown of STYK1 remarkably attenuated viability and clonality in Hep-2 cells, while overexpression of STYK1 achieved the opposite trends in AMC-HN-8 cells (p<0.05). TGF-β1 was confirmed to be the direct target binding STYK1, whose expression level was negatively regulated by STYK1. TGF-β1 was significantly down-regulated in LSCC tissues (p<0.05). Meanwhile, its low expression predicted significantly poor prognosis of LSCC patients. In addition, TGF-β1 was responsible for STYK1-regulated malignant progression of LSCC. STYK1 is upregulated in LSCC and is closely associated with T stage and poor prognosis. Furthermore, STYK1 promotes the proliferative ability of LSCC cells through targeting TGF-β1, thus aggravating the malignant progression of LSCC.

Serum levels of microRNA-21 and microRNA-10a can predict long-term prognosis in laryngeal cancer patients: a multicenter study

BackgroundLaryngeal squamous cell carcinoma (LSCC) ranks as the second most common subtype of squamous cell sarcoma (SCC) of the head and neck. In clinical practice, more tools are required to assist doctors in identifying patients who are at risk of poor prognosis at an early stage. Many studies have shown that microRNA-21 plays an important role in LSCC. MicroRNA-10a has also been found to be differentially expressed in squamous cell carcinomas of the head and neck. This study aimed to test whether circulating microRNA-21 and microRNA-10a could serve as prognostic predictors for patients with LSCC.MethodsBetween May 2009 and Jan 2015, laryngeal cancer patients who were treated at three medical centers were enrolled based on inclusion and exclusion criteria. Baseline blood samples were obtained before surgery, chemotherapy, or radiotherapy. MicroRNA array was used to detect the microRNAs that were differently expressed between the LSCC patients and healthy controls. The serum levels of selected microRNAs were tested in each patient using qRT-PCR. Clinical information was acquired from the patients’ clinic or in-hospital records and all of the patients were followed-up.ResultsA total of 236 patients (average age, 59.4±10.7 years; male, 223; 33.5% with supraglottic LSCC, and 66.5% with glottis LSCC) were enrolled into the final analysis. MicroRNA array showed that the serum level of microRNA-21 was higher and that of microRNA-10a was lower in the LSCC patients compared with the healthy controls. Patients at T I/II stage, N0 stage, M0 stage had lower serum level of microRNA-21 and higher serum level of microRNA-10a. Serum level of microRNA-21 and microRNA-10a both differed significantly between patients with LSCC of well, moderate and poor differentiation. Cox proportional hazards model revealed that T stage, N stage, M stage, age, and ratio of serum level of microRNA-21 and microRNA-10a were risk factors associated with 5-year survival rate in patients with LSCC. Kaplan Meier survival analysis revealed that lower baseline ratio of serum level of microRNA-21 and microRNA-10a predict better 5 years survival. The receiver-operating characteristic curve demonstrated the ratio had a good ability to discriminate patients survive at the end of five years follow-up, with an area under the curve (AUC) of 0.8965 (95% CI: 0.7849–0.9811).ConclusionsThe serum levels of microRNA-21 and microRNA-10a are associated with long-term prognosis in laryngeal cancer patients. Lower serum level of microRNA-21 and higher serum level of microRNA-10a predict better prognosis in LSCC patients.

Lack of miR-1246 in small extracellular vesicle blunts tumorigenesis of laryngeal carcinoma cells by regulating Cyclin G2.

Small extracellular vesicle (sEV) has precise impacts on tumor microenvironment and play vital functions in intercellular interaction. However, the functional role of sEV miRNA on laryngeal squamous cell carcinoma (LSCC) is largely unresolved. Here, the expression of miR-1246 in LSCC tissues and plasma sEV was examined. The internalization ability of sEV was determined by uptake assay. Then, the source and purity of sEV were checked through RNase and/or pharmacological inhibitors application. The invasion, migration, proliferation, and cell cycle assays were used to determine the altered abilities of miR-1246 in sEV in LSCC. Finally, target gene of miR-1246, Cyclin G2 (CCNG2), was stained immunohistochemically. In addition, the relationship between CCNG2 and clinicopathological features of patients was analyzed. We found that miR-1246 was higher in LSCC tissues and plasma sEV. MiR-1246 was enriched in sEV rather than soluble form. SEV could be internalized into adjacent cells. Lack of miR-1246 in sEV abrogated the tumorigenesis of LSCC. Furthermore, CCNG2 knockdown arrested the cell cycle and correlated to clinicopathological features and prognosis of LSCC patients. Taken together, we found that the function of sEV miR-1246 by regulating CCNG2 is responsible for LSCC advancement with emphasis on the main source of miR-1246 mainly root in sEV rather than in soluble form.

PLOD2 contributes to drug resistance in laryngeal cancer by promoting cancer stem cell-like characteristics

BackgroundAdvanced stage laryngeal squamous cell carcinoma (LSCC) presents a poor prognosis; thus, there is a great need to identify novel prognostic molecular markers. Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 2 (PLOD2) is thought to be a novel prognostic factor in several cancers, but its role in LSCC remains unknown. Cancer stem cells (CSCs) are responsible for most instances of tumor recurrence and the development of drug resistance and have been proven to be present in head and neck cancers. Our preliminary study indicated that PLOD2 was elevated in LSCC tissues; therefore, we hypothesized that PLOD2 is related to the prognosis of LSCC patients and aimed to explore the role and underlying mechanism of PLOD2 in LSCC.MethodsWe validated the prognostic role of PLOD2 in 114 LSCC patients by immunohistochemistry. Stable PLOD2-overexpressing Hep-2 and FaDu cells were established and assessed by molecular biology and biochemistry methods both in vitro and in vivo.ResultsWe confirmed that PLOD2 overexpression was correlated with poor prognosis in LSCC patients. PLOD2 overexpression strengthened the CSC-like properties of Hep-2 and FaDu cells, activated the Wnt signaling pathway and conferred drug resistance in LSCC in vitro and in vivo.ConclusionsWe found that PLOD2 could serve as a prognostic marker in patients with LSCC and confer drug resistance in LSCC by increasing CSC-like traits; in addition, a Wnt-responsive CSC pathway was identified.

Abstract 1261: Targeting Wee1 in laryngeal squamous cell carcinoma

Abstract WEE1 is a tyrosine kinase that regulates G2/M cell cycle checkpoint and frequently overexpressed in various tumors. However, the expression and clinical significance of WEE1 in human laryngeal squamous cell carcinoma (LSCC) are still unknown. In this study, we found that WEE1 was highly expressed in LSCC tissues compared with adjacent normal tissues. Importantly, overexpression of WEE1 was correlated with T stages, lymph node metastasis, clinical stages and poor prognosis of LSCC patients. Furthermore, inhibition of WEE1 by MK-1775 induced cell growth inhibition, G2/M cell cycle arrest and apoptosis with the increased intracellular reactive oxygen species (ROS) levels in LSCC cells. Pretreatment with ROS scavenger N-acetyl-L-cysteine could reverse MK-1775-induced ROS accumulation and cell apoptosis in LSCC cells. MK-1775 also inhibited the growth of LSCC xenografts in nude mice. Altogether, these findings suggest that WEE1 is a potential therapeutic target in LSCC, and inhibition of WEE1 is the prospective strategy for LSCC therapy. Note: This abstract was not presented at the meeting. Citation Format: Meng-Ling Yuan, Qi-Wei Jiang, Yang Yang, Jia-Rong Huang, Zhi Shi. Targeting Wee1 in laryngeal squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1261.

MiR-154 inhibits the growth of laryngeal squamous cell carcinoma by targeting GALNT7.

MicroRNAs are critical regulators of the development and progression of laryngeal squamous cell carcinoma (LSCC). However, the role of microRNA-154 (miR-154) in the development and progression of LSCC has not been clarified. We found that down-regulated miR-154 expression in LSCC tissues was associated with poorer prognosis in LSCC patients. MiR-154 over-expression inhibited the proliferation, clonogenicity, and migration of LSCC cells and induced cell cycle arrest, which were reversed by miR-154 inhibition. MiR-154 targeted GALNT7 expression by reducing GALNT7-regulated luciferase activity in LSCC cells while up-regulating GALNT7 mRNA transcription in LSCC tissues and cells. GALNT7 silencing significantly attenuated the proliferation, clonogenicity, and migration of LSCC cells and induced cell cycle arrest. Finally, intravenous treatment with lentivirus for miR-154, but not scrambled control miRNA, significantly restrained the growth of implanted LSCC Hep-2 tumors and decreased the tumor mass by reducing GALNT7 expression in mice. Therefore, miR-154 may serve as a novel prognostic marker and therapeutic target for LSCC.

The Clinical Signification of Claudin-11 Promoter Hypermethylation for Laryngeal Squamous Cell Carcinoma.

BackgroundClaudin-11 (CLDN11) is frequently silenced by its promoter hypermethylation. Previous studies have shown that CLDN11 promoter hypermethylation is a potential biomarker for diagnosing various cancers. The aim of this study was to investigate CLDN11 promoter methylation and its potential relevance to clinicopathologic features and prognosis of patients with laryngeal squamous cell carcinoma (LSCC).Material/MethodsUsing the quantitative methylation-specific polymerase chain reaction (qMSP), CLDN11 promoter methylation was measured in 91 tumor tissues and their paired adjacent normal tissues, and the relationship between CLDN11 methylation and clinicopathologic features was evaluated. A receiver operating characteristic (ROC) curve was created to assess diagnostic values, and the Kaplan-Meier survival analysis was used to evaluate the association between CLDN11 methylation and prognosis of patients with LSCC.ResultsOur results showed significantly elevated promoter methylation of CLDN11 in tumor tissues compared to their adjacent tissues (p=1.227E-16). CLDN11 promoter methylation also increased in patients with lymph node metastasis (p=0.009), advanced clinical stage (p=9.26E-06) and higher T classification (p=0.003). The area under the ROC curve (AUC) of CLDN11 was 0.884 (95% CI=0.835–0.932, p<0.01). The Kaplan-Meier analysis indicated that high CLDN11 promoter methylation levels were associated with poor overall survival of LSCC patients (log-rank test, p=0.007).ConclusionsWe demonstrated that CLDN11 promoter hypermethylation is a frequent event in LSCC, and contributes to metastasis and progression of LSCC. Thus, CLDN11 could be a potential biomarker for diagnosis and prognosis of LSCC patients.

HSP47 is associated with the prognosis of laryngeal squamous cell carcinoma by inhibiting cell viability and invasion and promoting apoptosis.

Heat shock protein47 (HSP47) is a 47kDa collagen binding protein that has a close relationship with the development and progression of tumours. However, little is known concerning the expression profile of HSP47 in laryngeal squamous cell carcinoma (LSCC) patients and there is still insufficient data concerning the underlying mechanisms. The aim of the present study was to explore the expression of HSP47 in LSCC and provide an overview of its association with tumourigenicity and clinical prognosis. The expression of HSP47 in LSCC and adjacent non-cancerous laryngeal tissues was assessed via western blotting and immunohistochemical studies. The prognostic significance of HSP47 expression was analysed using a Kaplan-Meier survival curve. To investigate the influence of HSP47 on the viability, invasion and apoptosis of a LSCC cell line, we performed an invitro analysis with plasmid vectors and small interfering RNA (siRNA). Our results showed that HSP47 protein expression in the LSCC tissues was markedly decreased compared to that noted in the adjacent non-cancerous tissues, and low expression of HSP47 was correlated with poor prognosis in LSCC patients. Upregulation of HSP47 via plasmid vectors inhibited the proliferation, reduced the invasive ability, increased the sensitivity to cisplatin chemotherapy, promoted apoptosis, and induced the G1phase arrest of LSCC cells invitro. The expression of apoptosis-regulating proteins was also altered when HSP47 was upregulated, involving increased expression of cleaved caspase-7/-8/-9, PARP, and Bax and decreased expression of Bcl-2. Our present data suggest that HSP47 is an important prognostic factor and an attractive therapeutic target in LSCC due to its influence on the biological behaviour of LSCC cells.

HOXB9 Expression Correlates with Histological Grade and Prognosis in LSCC.

The purpose of this study was to investigate the HOX gene expression profile in laryngeal squamous cell carcinoma (LSCC) and assess whether some genes are associated with the clinicopathological features and prognosis in LSCC patients. The HOX gene levels were tested by microarray and validated by qRT-PCR in paired cancerous and adjacent noncancerous LSCC tissue samples. The microarray testing data of 39 HOX genes revealed 15 HOX genes that were at least 2-fold upregulated and 2 that were downregulated. After qRT-PCR evaluation, the three most upregulated genes (HOXB9, HOXB13, and HOXD13) were selected for tissue microarray (TMA) analysis. The correlations between the HOXB9, HOXB13, and HOXD13 expression levels and both clinicopathological features and prognosis were analyzed. Three HOX gene expression levels were markedly increased in LSCC tissues compared with adjacent noncancerous tissues (P < 0.001). HOXB9 was found to correlate with histological grade (P < 0.01) and prognosis (P < 0.01) in LSCC. In conclusion, this study revealed that HOXB9, HOXB13, and HOXD13 were upregulated and may play important roles in LSCC. Moreover, HOXB9 may serve as a novel marker of poor prognosis and a potential therapeutic target in LSCC patients.

LAMP1 expression is associated with malignant behaviours and predicts unfavourable prognosis in laryngeal squamous cell carcinoma

Lysosome-associated membrane protein-1 (LAMP1) has been suggested to play complicated roles in cancer development and metastasis. The aim of this study was to explore the expression of LAMP1 in laryngeal squamous cell carcinoma (LSCC) and investigate the relationship between LAMP1 expression and clinicopathological characteristics in LSCC patients. One-step quantitative reverse transcription PCR (qPCR) tests (20 fresh LSCC and non-cancerous tissue samples) and immunohistochemistry (IHC) analyses (137 paraffin-embedded LSCC and non-cancerous tissue samples) were performed to evaluate the LAMP1 expression in both mRNA and protein levels. Results showed that the expression of LAMP1 in LSCC tissues was significantly higher than that in non-cancerous tissues. Furthermore, the expression level of LAMP1 protein was statistically associated with lymph node metastasis and TNM stage. Results of Kaplan-Meier survival and Cox regression analyses revealed that LSCC patients with high LAMP1 cytoplasmic expression (p=0.044), high cytoplasmic+low mesenchymal expression of LAMP1 (H+L) (p=0.015) and histopathological grade (p=0.014) encountered poor overall survival. The data implied that high LAMP1 expression is associated with unfavourable prognosis in LSCC patients, and LAMP1 may be identified as a novel prognostic biomarker.

Clinical value of serum HMGB1 in diagnosis and prognosis of laryngeal squamous cell carcinoma.

Laryngeal squamous cell carcinoma (LSCC) is one of the most common malignancies in the head and neck region. Recently, aberrantly expressed high-mobility group box 1 (HMGB1) has received a great deal of attention as a potential biomarker for cancer diagnosis and prognosis. Therefore, the aim of this study was to estimate HMGB1 levels in serum in LSCC patients and healthy controls and evaluated the potential of serum HMGB1 as a noninvasive biomarker for diagnosis and prognosis in LSCC. Serum HMGB1 levels were analyzed in 71 LSCC patients and 50 healthy controls. The serum HMGB1 level was significantly higher in LSCC patients compared with the healthy controls (4.81 ± 2.33 vs. 3.21 ± 1.08 ng/mL, P < 0.001). High serum HMGB1 was significantly associated with T classification (P = 0.005), N classification (P = 0.002), and clinical stage (P = 0.001). The area under ROC curve was 0.716, and the sensitivity and specificity were 42.3 and 92.0%, respectively. The Kaplan-Meier plots showed that patients with high serum HMGB1 had a poorer overall survival than those with low serum HMGB1 (P = 0.036). Serum HMGB1 levels are significantly associated with the progression of LSCC. In this population, HMGB1 has a poor sensitivity, but a high specificity for the diagnosis of LSCC. Serum HMGB1 level has potential as a biomarker for the prognosis in LSCC patients.

MicroRNA-126 modulates the tumor microenvironment by targeting calmodulin-regulated spectrin-associated protein 1 (Camsap1)

Plasma miRNAs have been reported as biomarkers for various diseases. In this study, we investigated whether plasma concentrations of miR-126 may be useful as biomarkers for laryngeal squamous cell carcinoma (LSCC). We examined the function and mechanism of miR-126 in LSCC by using cell biology and molecular pathology techniques such as western blotting, quantitative PCR, IHC and IF. The expression of Camsap1 mRNA and protein is higher in cancer tissues compared to that in normal tissues. Both miR-126 and Camsap1 were related with the prognosis of LSCC patients. We found that miR-126 was able to inhibit LSCC partly by suppressing Camsap1 expression. In addition, Camsap1 expression induced microtubule formation and aggregation. This mechanism possibly explains why loss of miR-126 is frequently associated with tumor metastasis.

Expression TGM2 and BNIP3 have prognostic significance in laryngeal cancer patients receiving surgery and postoperative radiotherapy: A retrospective study

BackgroundThis study was designed to determine the pattern and correlation between expression of the HIF-1α transcriptional targets TGM2 and BNIP3 in laryngeal cancer, and investigate the association of BNIP3 and TGM2 with clinical outcome in laryngeal squamous cell carcinoma (SCC) patients receiving postoperative radiotherapy.MethodsImmunostaining with antibodies specific to BNIP3 and TGM2 was performed in formalin-fixed, paraffin-embedded specimens from 148 laryngeal SCC patients. BNIP3 and TGM2 expression was scored as high or low, based on the number of tumor cells stained and the staining intensity. All patients received postoperative radiotherapy. Patient follow up and clinicopathological data were compared using the Chi-squared test, univariate and multivariate analyses, and survival curves were generated using the Kaplan-Meier method and log-rank test.ResultsThe 3, 5 and 10-year overall survival rates (OS) for all patients were 77.7%, 71.6%, 56.4%, respectively. Primary tumor site, T stage, overall stage, lymph-node metastasis, BNIP3 expression and TGM2 expression were significant prognostic factors for OS in univariate analysis. Negative cervical lymph nodes, high BNIP3 expression and low TGM2 expression were independent prognostic factors of improved OS in multivariate analysis. BNIP3 expression correlates with TGM2 expression in laryngeal SCC (P = 0.012).ConclusionsThis study indicates that lymph-node metastasis, BNIP3 expression and TGM2 expression are independent prognostic factors in laryngeal SCC patients receiving postoperative radiotherapy. Further studies are required to investigate how BNIP3 and/or TGM2 influence the prognosis of laryngeal SCC patients treated with postoperative radiotherapy, and to determine how TGM2 and BNIP3 expression are regulated.