Primary Hypopharyngeal Squamous Cell Carcinoma Research Articles

Assessment and treatment strategies for occult contralateral lymph node metastasis in hypopharyngeal squamous cell carcinoma patients with ipsilateral node-positive necks.

Hypopharyngeal squamous cell carcinoma (HSCC) is an aggressive disease with poor prognosis, yet studies have largely been more qualitatively focused. Our study aims to quantitatively predict the risk of occult contralateral lymph node metastasis (cLNM) for HSCC patients with ipsilateral lymph node metastasis (iLNM). This will be based on pre- and post-operative indexes to guide the selection of prophylactic contralateral lymph node dissection (cLND) and postoperative adjuvant treatments. Multivariate analyses of 462 primary HSCC patients with iLNM showed that the age of patients, subregions of tumor, pathological T (pT) stage, ipsiateral MLS and metastatic lymph node number (MLN), and lymph nodal necrosis were independent cLNM risk factors. These were used to construct two nomograms that can effectively predict the contralateral neck involvement in HSCC patients with ipsilateral positive lymph nodes. The first nomogram (pre-model) provides quantitative assessment on the necessity of cLND, while the second nomogram (post-model) informs regions of interest for therapeutic radiation. Overall, patients deemed high-risk of cLNM by pre-model should receive cLND. Post-operation, patients deemed high-risk of cLNM by post-model should receive therapeutic radiation targeting contralateral neck lymph nodes, moderate-risk group warrants comparatively lower dose contralaterally, while low-risk group requires only follow-up.

Pparg may Promote Chemosensitivity of Hypopharyngeal Squamous Cell Carcinoma.

The upregulation of peroxisome proliferator-activated receptor gamma (PPARG) has been shown to increase the chemosensitivity of several human cancers. This study is aimed at studying if PPARG sensitizes hypopharyngeal squamous cell carcinoma (HSCC) in chemotherapeutic treatments and at dissecting possible mechanisms of observed effects. We integrated large-scale literature data and HSCC gene expression data to identify regulatory pathways that link PPARG and chemosensitivity in HSCC. Expression levels of molecules within the PPARG regulatory pathways were compared in 21 patients that underwent chemotherapy for primary HSCC, including 12 chemotherapy-sensitive patients (CSP) and 9 chemotherapy-nonsensitive patients (CNSP). In the CPS group, expression levels of PPARG were higher than that in the CNSP group (log‐fold‐change = 0.50). Structured text mining identified two chemosensitivity-related regulatory pathways driven by PPARG. In the CSP group, expression levels for 7 chemosensitivity-promoting genes were increased, while for 13 chemosensitivity suppressing the gene expression levels were decreased. Our results support the chemosensitivity-promoting role of PPARG in HSCC tumor cells, most likely by affecting both cell proliferation and cell motility pathways.

Dynamic prediction of cancer-specific survival for primary hypopharyngeal squamous cell carcinoma.

This study investigated a large cohort of patients to construct a predictive nomogram and a web-based survival rate calculator for dynamically predicting the cancer-specific survival of patients with primary hypopharyngeal squamous cell carcinoma (HSCC). Patients (n = 2007) initially diagnosed with primary HSCC from 2004 to 2015 were extracted from the Surveillance, Epidemiology, and End Results (SEER) database. All patients were randomly divided into the training and validation cohorts (1:1). The Lasso Cox regression model was applied to identify independent risk factors of cancer-specific survival for a predictive nomogram and a web-based calculator. The model was evaluated by concordance index, calibration, and decision curve analysis. Cancer-specific survival rates decreased with time, while 3-year conditional survival increased. Cancer-specific deaths evolved from relatively high within the first 3years to low thereafter. Age, race, T stage, N stage, M stage, surgery, radiotherapy, chemotherapy, and marital status were identified as independent risk factors. We constructed a predictive nomogram for survival and a web-based calculator ( https://linzhongyang.shinyapps.io/Hypopharyngeal/ ). Additionally, a prognostic risk stratification was developed according to nomogram total points. Patients with primary HSCC were found at a high risk of cancer-specific death during the first 3years, indicating that additional effective follow-up strategies should be implemented over the period. This is the first study to construct a predictive nomogram and a web-based calculator for all patients with HSCC.

A prognostic nomogram for predicting the long-term survival outcome of hypopharyngeal squamous cell carcinoma patients after tumour resection to assist the decision-making of postoperative adjuvant treatment

BackgroundSurgery remains the most common therapeutic strategy for Chinese patients with hypopharyngeal squamous cell carcinoma (HSCC), yet the indications for postoperative adjuvant treatment (POAT) remain unclear. MethodsWe retrospectively analysed 385 patients with primary HSCC in our hospital between 2003 and 2014. Patients that received pharyngectomy without POAT were enrolled in this study for developing a nomogram that predicts their survival outcome. ResultsMultivariate analyses showed that the tumour size, oesophagal invasion, extracapsular spread or internal jugular vein adhesion, thyroid gland invasion, and the number of lymph node metastases (≤3 or >3) were significantly correlated to the overall survival (OS) of the patient and were included as risk factors in the nomogram. The C-index was 0.768 (95% CI, 0.719–0.817) in development cohort and 0.767 (95% CI, 0.753–0.781) in validation cohort. A calibration curve was also conducted and was found favourable. The patients were stratified into three groups based on their nomogram scores. In the high-risk group, patients that received POAT had a better OS than those that received only surgery. In the moderate-risk group, POAT did not show any significant association with the OS. However, patients in the low-risk group that received POAT showed a worse OS than those without. ConclusionThe newly-developed nomogram can effectively predict the survival outcome of patients with HSCC. According to the novel stratification criteria created, patients stratified as high-risk could benefit from POAT, while those in the low-risk group are advised not to receive POAT as this correlates with a worse OS.

Loss of CDH1 promotes the metastasis of hypopharyngeal squamous cell carcinoma through the STAT3-MMP-9 signaling pathway.

BackgroundDistant metastasis is the major cause of death in patients with hypopharyngeal squamous cell carcinoma (HSCC). CDH1 is correlated with tumor invasion and metastasis; however, its function in HSCC remains unclear.MethodsWe used immunohistochemistry (IHC) staining to evaluate the expression of CDH1 in 31 and 78 specimens from primary HSCC patients with and without postoperative lung metastases respectively. Sulforhodamine B (SRB) and CCK-8 assays were used to test the proliferation of HSCC cells. Motility of HSCC cells was investigated by migration and invasion assays. Western blot analysis was used to measure the levels of CDH1 and other proteins.ResultsWe found that the low expression of CDH1 was significantly associated with postoperative lung metastasis in HSCC (P<0.001). Moreover, CDH1 was reduced concomitantly with the upregulation of MMP-9 in the same HSCC sample. Further mechanistic investigation showed that silencing CDH1 elevated the level of MMP-9, which was coupled with the phosphorylation of STAT3. Subsequently, inhibiting STAT3 either by siRNA transfection or by pharmacological suppression with AG490 attenuated MMP-9 upregulation and prevented the enhanced proliferation and invasion caused by CDH1 loss in FaDu cells.ConclusionsCDH1 plays vital roles in HSCC metastasis and might serve as a potential therapeutic target for the clinical treatment of HSCC.

Low expression of AIM2 combined with high expression of p‑STAT3 is associated with poor prognosis in hypopharyngeal squamous cell carcinoma.

Studies indicate that the AIM2 inflammasome plays an important role in tumor occurrence and development. The present study examined the prognostic significance of AIM2 expression and the correlation between AIM2 expression and p‑STAT3 expression in hypopharyngeal squamous cell carcinoma (HSCC), which remain unknown. AIM2 and p‑STAT3 were detected using immunohistochemistry (IHC) in 111 paraffin‑embedded tissue samples from primary HSCC sites and 20 samples from matched adjacent non‑malignant sites. Western blotting was utilized to investigate AIM2, p‑STAT3 and total STAT3 in 5 pairs of fresh tissue samples. Western blotting indicated that AIM2 expression was significantly lower (P<0.05), but that p‑STAT3 levels and the p‑STAT3/STAT3 ratio were higher (P<0.01) in HSCC tissues than in adjacent normal hypopharyngeal tissues. IHC analysis of the 111 HSCC samples revealed low AIM2 and high p‑STAT3 expression in 48 (43.2%) and 58 (52.3%) samples, respectively. High AIM2 expression and low p‑STAT3 expression were detected in 13 (75%) and 18 (90%) adjacent normal hypopharyngeal tissue samples, respectively. In the HSCC samples, low AIM2 expression was closely related to lymph node metastasis and intravascular tumor thrombus (P<0.05). Kaplan‑Meier survival curves revealed that low AIM2 levels were strongly associated with poor survival for HSCC patients (P<0.0001). Additionally, Cox proportional hazards regression models indicated that low AIM2 expression significantly predicted poor prognosis for HSCC patients (P<0.0001), and multivariate analysis revealed that AIM2 expression could be an independent prognostic factor for HSCC patients (P<0.0001). Furthermore, AIM2 expression was negatively correlated with p‑STAT3 expression in the HSCC tissue samples, and combined analysis revealed that patients with low AIM2 and high p‑STAT3 levels had the worst survival rate. Moreover, receiver operating characteristic (ROC) curve analysis confirmed that AIM2 was predictive of specific survival in all HSCC patients [area under the curve (AUC)=0.7160]. In conclusion, our data suggested that for HSCC patients, AIM2 and p‑STAT3 expression detected via IHC could serve as a biomarker to predict tumor progression and as an independent prognostic factor.

Endoscopic screening for upper gastrointestinal second primary malignancies in patients with hypopharyngeal squamous cell carcinoma

Objective: To evaluate the usefullness of flexible esophagoscopy and chromoendoscopy with Lugol's solution in the detection of synchronous esophageal neoplasm in patients with hypopharyngeal squamous cell carcinoma (HSCC). Methods: A retrospective review of 96 cases with HSCC that received surgical treatment from March 2016 to March 2017 was accomplished. In these patients, the site of origin were pyriform sinus (n=75), posterior pharyngeal wall (n=11) and postcricoid (n=10). Esophagoscopy was prospectively performed on all patients before treatment for HSCC. All patients underwent conventional white-light endoscopic examination with Lugol chromoendoscopy and narrow band image. Suspicious areas of narrow band image or Lugol-voiding lesions were observed and biopsied. The treatment strategy of primary HSCC was modified according to the presence of synchronous esophageal squamous cell neoplasms by a multidisciplinary approach. Results: Ninety-six patients were enrolled (age ranging from 37-80 years). All patients did not have previous treatment.Histopathological analysis revealed middle to high-grade dysplasia in 5 cases, Tis cancer in 5 cases, cancer in 16 cases and inflammation or normal findings in the others. Four cases were treated with endoscopic submucosal dissection before hypopharygeal surgery, 3 cases with lower esophageal cancers were treated with gastric pull-up combined with free jejunal flap after total circumferential pharyngolaryngectomy (TCPL) and certical esophagectomy, and 14 cases were treated with TCPL, total esophagectomy and gastric pull-up. Conclusions: Esophagoscopy is a feasible and justified procedure in HSCC cases as it enhances the detection of premalignant lesion or second primary cancer. Routine esophagoscopy for detecting synchronous second primary tumor should be recommended for patients with HSCC. The treatment strategy for primary HSCC is modified according to the presence of synchronous second primary tumor.

Prognostic impact of perineural invasion in hypopharyngeal squamous cell carcinoma

Objectives: The aim of this study was to evaluate the role of PNI in HPSCC.Methods: The medical records of 105 patients who underwent surgery-based treatment for HPSCC were reviewed. Clinicopathologic parameters including disease-specific survival were correlated with PNI.Results: PNI was identified in 27 of the 105 (25.7%) cases of HPSCC. Correlation analysis demonstrated that PNI in HPSCC was significantly correlated with pN classification (10.3% in N0/N1 vs 34.8% in N2/N3, p = 0.006). Patients with PNI had decreased 5-year disease-specific survival with borderline significance (p = 0.065). In a sub-set of 31 patients who did not receive post-operative radiotherapy, PNI was determined to be a significant prognostic predictor (p = 0.033). In multivariate analysis, extracapsular invasion was the only independent prognostic factor for disease-specific survival (p = 0.001).Conclusion: Perineural invasion (PNI) should be considered an independent predictor for cervical lymph node involvement. PNI status in primary hypopharyngeal squamous cell carcinoma (HPSCC) specimens should be considered in decisions concerning adjuvant radiotherapy.

Abstract 3118: Identification of tumor suppressive microRNA-451a in hypopharyngeal squamous cell carcinoma based on microRNA expression signature

Abstract Background: Hypopharyngeal squamous cell carcinoma (HSCC) has a very poor prognosis because of its high rates of regional and distant metastasis. In spite of considerable advances in multimodality therapy, including surgery, radiotherapy and chemotherapy, the overall survival rate for patients with HSCC is only 15% - 45%. Identification of differentially expressed miRNAs and their regulated molecular targets in tumor cells might enhance our understanding of the molecular mechanisms of metastasis in human cancers. Recent our microRNA signature revealed that miR-451a was downregulated in HSCC clinical specimens, suggesting that it might be tumor suppressive microRNA. The aim of study was to investigate the functional significance of miR-451a and to identify the molecular targets regulated by miR-451a in HSCC. Method: XTT, migration, invasion assay were performed after transfection with miR-451a in FaDu (derived from a primary HSCC) and SAS (derived from a primary tongue SCC). Genome-wide gene expression analysis were used to analyze the candidate genes regulated by miR-451a. The expression levels of miR-451a and ESDN/DCBLD2 were analyzed by qRT-PCR in HSCC clinical specimens and cell lines. The luciferase reporter assay was used to identify the actual binding site of miR-451a in target gene. We performed qRT-PCR and Western blotting in two cell lines to investigate whether restoration of miR-451a altered ESDN/DCBLD2 gene and protein expression. Result: The expression levels of miR-451a was significantly lower in clinical HSCC specimens and cell lines compared with non-cancerous specimens. Restoration of miR-451a inhibited cancer cell migration and invasion in two cell lines. Genome-wide gene expression analysis of miR-451a transfectant showed that five genes (SPC25, MIF, ESDN/DCBLD2,C4orf46 and AKR1B1) were downregulated. Among these candidate genes, we focused on ESDN/DCBLD2 because it was the most significantly upregulated in cancer tissues, and was shown a negative correlation between the expression of miR-451a in Spearman's rank test. Luciferase reporter assay showed that ESDN/DCBLD2 was directly regulated by miR-451a in SAS. The mRNA and protein expression levels of ESDN/DCBLD2 were significantly repressed in miR-451a transfectant. Silencing of ESDN/DCBLD2 inhibited cell migration and invasion significantly in two cell lines. Conclusion: These data showed that tumor suppressive miR-451a inhibits cancer cell migration and invasion through regulating ESDN/DCBLD2. The identification of novel target oncogenes regulated by miR-451a might lead to a better understanding of HSCC metastasis and the development of new therapeutic strategies to treat this disease. Citation Format: Ichiro Fukumoto, Takashi Kinoshita, Toyoyuki Hanazawa, Naoko Kikkawa, Takeshi Chiyomaru, Hideki Enokida, Noriko Yamamoto, Yusuke Goto, Rika Nishikawa, Masayuki Nakagawa, Yoshitaka Okamoto, Naohiko Seki. Identification of tumor suppressive microRNA-451a in hypopharyngeal squamous cell carcinoma based on microRNA expression signature. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3118. doi:10.1158/1538-7445.AM2015-3118

Aberrant Expression of PHLPP1 and PHLPP2 Correlates with Poor Prognosis in Patients with Hypopharyngeal Squamous Cell Carcinoma

The PHLPP (pleckstrin homology [PH] domain leucine rich repeat protein phosphatase) family, which represents a family of novel Ser/Thr protein phosphatases, is composed of 2 members: PHLPP1 and PHLPP2. PHLPPs partake in diverse cellular activities to exhibit their antitumor and metastasis suppressor functions. It is necessary to investigate the expression patterns of PHLPP1 and PHLPP2 in hypopharyngeal squamous cell carcinomas (HSCCs) and clarify their clinical significance. A total of 138 patients with primary HSCC who underwent curative surgical treatment as an initial treatment were enrolled in this study. A total of 138 HSCC specimens and 64 adjacent noncancerous mucosal epithelial tissues were collected. The expression levels of PHLPP1 and PHLPP2 were examined by quantitative reverse transcription polymerase chain reaction and immunohistochemistry assays. Correlations between clinicopathological parameters of the patients were further evaluated. PHLPP1 and PHLPP2 mRNA transcript levels were significantly lower in tumor samples than in paired adjacent nontumor mucosae (P<0.0001, both). Positive correlations were observed between the mRNA levels of PHLPP1 and PHLPP2 in HSCC tissues (correlation coefficient r = 0.678, P<0.001) and in adjacent nontumor mucosae (r = 0.460, P<0.001). The majority of the noncancerous tissues showed high expression levels of PHLPP1 (87.5%, 56/64) and PHLPP2 (85.9%, 55/64). However, the expressions of PHLPP1 and PHLPP2 were significantly decreased in 83.3% (115/138) and 82.6% (114/138) of tumor tissues, respectively (P<0.0001, both). The expressions of both PHLPP isoforms were significantly related to the tumor clinical stage, differentiation, and cervical lymph node metastasis (P<0.05, all). It was PHLPP1 but not PHLPP2 that was significantly related to the tumor T stage. Low PHLPP1 and PHLPP2 expressions were associated with poor overall survival (OS) in HSCC patients (P = 0.004, P = 0.008, respectively). Multivariate analysis revealed that PHLPP1 was an independent prognostic factor for OS. This study indicates that, in HSCC, aberrant expressions of PHLPP1 and PHLPP2 are common events, and loss of PHLPPs might identify patients with poor prognostic outcomes.

Cyclin D1 overexpression in primary hypopharyngeal carcinomas.

Hypopharyngeal squamous cell carcinomas (HPCS) are associated with an extremely poor prognosis. Generally, conventional clinicopathologic factors have only limited value as prognostic factors for this malignancy. It is therefore clinically important to identify new prognostic factors that accurately reflect the biologic aggressiveness of this malignancy. The amplification and overexpression of the cyclin D1 protooncogene have been reported in a variety of malignancies, and are thought to be related to tumor progression. Based on this phenomenon, the authors immunohistochemically evaluated overexpression of the cyclin D1 gene in 42 cases of primary HPCS. In addition, the immunohistochemical staining of the proliferation marker MIB-1 (Ki-67 antibody) was also performed. Formalin fixed, paraffin embedded biopsy specimens obtained prior to treatment were examined. Cyclin D1 and Ki-67 were detected using monoclonal antibodies by means of the streptavidin-biotin method. The relationship between cyclin D1 overexpression and the stage, histologic grade, presence of lymph node metastases, proliferation index, and survival was then statistically analyzed. The correlation between the proliferation index, other clinicopathologic factors, and survival was also evaluated. Twenty-three (54.8%) HPCS specimens showed a 20% or greater immunoreactivity for cyclin D1. Cyclin D1 overexpression was related to cervical lymph node metastases (P = 0.037) but not to clinical stage, histologic grade, or the proliferation index. Cyclin D1 negative tumors were associated with a significantly better prognosis (P = 0.023), particularly in patients who underwent multimodality treatment. Finally, the MIB-1 labeling index showed no correlation with either the clinicopathologic parameters or overall survival. Based on these findings, cyclin D1 immunohistochemical staining is considered to be useful, not only as a prognostic factor for HPCS, but also as a means of determining the optimum treatment for each individual patient. Conversely, the MIB-1 labeling index appears to have no clinical significance in HPCS.