Anlotinib suppressed tumor cell proliferation and migration in hypopharyngeal carcinoma

Abstract Objective The purpose of this study is to study the in-vitro effects of multitarget inhibitor anlotinib on hypopharyngeal cancer cell proliferation and cell migration, and the underlying mechanism, which will provide new drug choices for hypopharyngeal cancer treatment. Methods The Hypopharyngeal cancer Fadu cells were treated with anlotinib at a concentration of 0, 5, and 10 μmoL/L, respectively. Cell counting kit-8 and the colony-forming assay were used to detect the inhibition of cell proliferation. Wound-healing assay and transwell assay were used to detect the migration and invasion ability of cells. Flow cytometry was used to detect the effects of anlotinib on cell cycle and apoptosis. RT-qPCR and Western blot were used to measure gene expression levels. Results CCK-8 and colony-forming assay showed that anlotinib could significantly inhibit cell proliferative activity. Wound-healing assay and transwell assay showed that anlotinib could inhibit cell migration and scratch. These results showed that anlotinib has obvious antitumor activity. Flow cell cycle experiment showed that anlotinib could promote Fadu cell apoptosis and block the G2/M phase for inhibiting cell proliferation. In addition, anlotinib decreased the expression of HIF-1α. Conclusions Anlotinib has an excellent suppressing effect on the proliferation, migration, and invasion of hypopharyngeal cancer Fadu cells in-vitro. Moreover, it can play an anti-tumor role through blocking cell cycle G2/M and promoting apoptosis, which may be related to the decrease of HIF-1a expression. Our study would provide a potential treatment method for patients with hypopharyngeal cancer. Level of evidence: Level 3.

The role and clinical significance of microRNA-29a-3p in the development of hypopharyngeal carcinoma

Abstract Objective MicroRNA-29a-3p has been reported in a variety of cancers, but its role in hypopharyngeal cancer remains unclear. This study was to determine the role of microRNA-29a-3p in the occurrence and development of hypopharyngeal cancer. Methods 40 patients with hypopharyngeal cancer who underwent surgery in the Affiliated Hospital of Jining Medical University from April 2013 to November 2017 were selected for this study. The cancer tissue samples of the patients were collected, and the patients were followed up for three years. The expression of microRNA-29a-3p in tissue samples was detected by in situ hybridization with fluorescent probe, and the relationships among microRNA-29a-3p and clinicopathological factors, postoperative recurrent-metastasis, survival time were studied. Immunohistochemical was used to detect the expression of Ki67 and E-cadherin in tissue samples. Results Combined with HE staining results showed that microRNA-29a-3p expression was relatively high in non-cancer tissue cells (red blood cells and fibroblasts in tumor interstitial vessels), but was relatively low in cancer tissue and cells. According to the follow-up data of 40 patients with hypopharyngeal cancer, tumor size, T-stage, tumor differentiation, postoperative recurrent-metastasis of hypopharyngeal cancer patients were significantly negatively correlated with microRNA-29a-3p (p< 0.05). Immunohistochemica results further confirmed that microRNA-29a-3p was negatively correlated with the expression of Ki67 and E-cadherin. The survival time of patients positively related with microRNA-29a-3p expression (p< 0.05). Moreover, ROC curve analysis showed that the area under the curve of the combined detection of miRNA-29a-3p+Ki67+E-cadherin was larger than that of the single detection of the three indexes. Conclusions The expression of microRNA-29a-3p is closely related to the occurrence, development and prognosis of hypopharyngeal cancer, and it affects the proliferation and invasion. This indicates that microRNA-29a-3p serves as a therapeutic target for the occurrence and development of hypopharyngeal cancer. The evidence of study designs of this study is IV using "Oxford Centre for Evidence-Based Medicine 2011 Levels of Evidence".

Clinical characteristics and efficacy of second primary malignancies in hypopharyngeal carcinoma: an analysis of 216 real-world cases / 中华放射肿瘤学杂志

Objective:To evaluate the incidence, clinical characteristics and prognosis of second primary malignancies (SPMs) among patients with hypopharyngeal carcinoma (HPC) in real-world analysis.Methods:A total of 594 HPC patients admitted to Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College from 2010 to 2018 were retrospectively analyzed.The incidence and clinical characteristics of HPC patients complicated with SPMs were analyzed. Clinical efficacy was compared among different groups.Results:With a median follow-up time of 66.9 months, SPMs were present in 36.4% (216/594) of HPC patients: 22.2% (132/594) were synchronous and 14.1% (84/594) were metachronous. The upper aerodigestive tract was the most common involved region. Compared with patients without SPMs, patients with synchronous and metachronous carcinoma in situ had similar 5-year overall survival (OS) of 42.2% vs. 44.5% ( P=0.958) and 62.2% vs. 44.5% ( P=0.240), respectively. Patients with synchronous invasive SPMs had a worse 5-year OS of 27.2% vs. 44.5% in their counterparts without SPMs ( P=0.001). Patients with metachronous invasive SPMs had similar 5-year OS of 50.2% vs. 44.5% in their counterparts without SPMs ( P=0.587). SPMs accounted for 42.5% of total death in metachronous invasive SPMs group. Conclusions:Patients with HPC have a high probability of developing SPMs. Moreover, the incidence of complicated with esophageal/gastric carcinoma in situ or metachronous SPMs exerts no effect on prognosis, while the occurrence of synchronous SPMs significantly affectes the prognosis of patients. However, the incidence of SPMs is still one of the main death causes in metachronous invasive SPMs group.

Analysis of clinical efficacy of postoperative low-dose radiotherapy in patients with advanced hypopharyngeal cancer without high-risk factors / 中华放射肿瘤学杂志

Objective:To evaluate the efficacy of low-dose radiotherapy in patients with advanced hypopharyngeal cancer without high-risk factors.Methods:Clinical data of 235 patients diagnosed with advanced hypopharyngeal cancer treated in Department of Head and Neck Surgery of Shandong Provincial ENT Hospital from December 2013 to August 2018 were retrospectively analyzed. All patients were divided into two groups: low-dose radiotherapy group (50 Gy, n=158) and high-dose radiotherapy group (>60 Gy, n=77). Clinical baseline characteristics, treatment, follow-up and survival of patients were collected. Survival curve was delineated using the Kaplan-Meier method, and the differences in survival between two groups were calculated using the log-rank test. Clinical baseline characteristics between two groups were compared by χ2 test. Univariate and multivariate analyses of prognostic factors were conducted by logistic regression model. Results:The median follow-up time was 45 months (5-94 months). The 3-year overall survival (OS) rate of the whole group was 68.5%, and 70.3% and 64.9% in the low-dose and high-dose groups, respectively ( P=0.356). The 3-year progression-free survival (PFS) rate of the whole group was 64.3%, and 65.8% and 61.0% in the low-dose and high-dose groups, respectively ( P=0.361). Univariate analysis showed that T stage, N stage, lesion location and degree of pathological differentiation significantly affected clinical prognosis (all P<0.05), whereas there was no significant relationship between age, sex, radiotherapy dose, interval between surgery and radiotherapy and survival. Multivariate analysis showed that T stage, N stage and the degree of pathological differentiation were the independent prognostic factors (all P<0.05) of the 3-year OS and PFS. Sex, radiotherapy dose and interval between surgery and radiotherapy were not correlated with OS and PFS. Conclusion:This study showed that for hypopharyngeal cancer patients without positive surgical margins and extracapsular extension, postoperative radiotherapy at a dose of 50 Gy given to tumor bed and selective lymph node drainage area does not compromise local disease control and OS.

Value of low-energy virtual monochromatic images of dual-energy CT in the evaluation of tumor visibility and T staging in hypopharyngeal squamous cell carcinoma / 中华放射学杂志

Objective:To investigate the value of low-energy virtual monoenergetic image (VMI) at 45 keV in visualizing the primary tumor and T staging of hypopharyngeal squamous cell carcinoma.Methods:The clinical and imaging data of 58 patients with hypopharyngeal squamous cell carcinoma from April 2018 to January 2020 at Eye & ENT Hospital, Fudan University were analyzed retrospectively. All the patients underwent a venous phase contrast-enhanced dual-source dual-energy CT scan before treatment. The VMI at 45 keV and standard linearly blended image (30% 80 kV+70% 140 kV) were acquired from dual-energy post-processing software. One senior radiologist and one junior radiologist independently assessed the visibility of the tumor on the 45 keV VMI and standard linearly blended image using a 5-point Likert rating scale. Furthermore, the senior radiologist assessed the visibility of the tumor at each subsite (piriform fossa, posterior pharyngeal wall, postcricoid region) and determined the invasion depth of the tumor (extension to esophagus, invasion to strip muscles and prevertebral muscles) and performed the T staging of the primary tumor using the two sets of images blindly. The accuracy of T staging was calculated, using pathological T staging (surgical cases) or clinical T staging (non-surgical cases) as the gold standard. The image scores of the two sets of images were compared using Wilcoxon rank sum test. McNemar-Bowker test was used to compare the accuracy of T staging using the two sets of images.Results:The overall image scores of the 45 keV VMI and standard linearly blended image from the senior radiologist were 3.5 (3, 4) and 3 (2, 3) respectively ( Z=-7.03, P<0.001), and the scores from the junior radiologist were 3 (3, 4) and 2 (2, 3) ( Z=-6.93, P<0.001). The scores of the 45 keV VMI were significantly higher than those of the standard linearly blended image in visualizing tumors in the piriform fossa, posterior pharyngeal wall, and postcricoid region, as well as in detecting invasion to the strip muscles ( P<0.05). There was no significant difference in the scores of the two sets of images in determining whether the tumor extended to esophagus or invaded prevertebral muscles ( P>0.05). Referring to pathological and clinical T stage, the accuracy of T staging determined by the 45 keV VMI and standard linearly blended image was 87.9% (51/58) and 81.0% (47/58) respectively, and the difference was not significant (χ 2=3.33, P=0.189). Conclusions:The 45 keV VMI is superior to the standard linearly blended image in visualizing tumors and detecting invasion to the strip muscles of hypopharynx squamous cell carcinoma. However, the accuracy of determining T staging using 45 keV VMI is slightly improved than that of standard linearly blended image, and the difference is not statistically significant. In determining whether the tumor extends to esophagus or invades prevertebral muscles, 45 keV VMI shows no significant advantage over standard linearly blended image.

Clinical prognosis and influencing factors of patients with hypopharyngeal squamous cell carcinoma treated by salvage surgery / 国际肿瘤学杂志

Objective:To investigate the clinical prognosis and influencing factors of patients with hypopharyngeal squamous cell carcinoma treated by salvage surgery.Methods:A total of 78 patients with hypopharyngeal squamous cell carcinoma underwent salvage surgery in Jincheng Second People's Hospital of Shanxi Province from January 2017 to January 2022 were included retrospectively. Postoperative complications were recorded. Logstic regression analysis was used to evaluate the influencing factors of survival after 5 years of salvage surgery, and receiver operating characteristic (ROC) curve was drawn to analyze the predictive value of various influencing factors on the survival of patients with hypopharyngeal squamous cell carcinoma after 5 years of salvage surgery.Results:The incidence of complications after salvage surgery in 78 patients was 21.79% (17/78) . The median total survival time was 20.5 months. There were 21 deaths 5 years after operation. Univariate analysis showed that the age of salvage surgery ( χ2=30.25, P<0.001) , location of recurrent tumor ( χ2=8.72, P=0.013) , surgical margin status ( χ2=6.93, P=0.008) , depth of tumor invasion ( χ2=8.31, P=0.004) and whether to accept radiotherapy (chemotherapy) after salvage operation ( χ2=4.24, P=0.040) were all related to the survival of patients with hypopharyngeal squamous cell carcinoma after 5 years of salvage surgery. Multivariate analysis showed that the status of the surgical margin ( OR=26.26, 95% CI: 4.58-150.62, P<0.001) , the depth of tumor invasion ( OR=14.03, 95% CI: 3.04-64.70, P<0.001) and whether to accept radiotherapy (chemotherapy) after the salvage surgery ( OR=7.73, 95% CI: 1.68-35.54, P=0.008) were independent factors affecting the survival of patients with hypopharyngeal squamous cell carcinoma after 5 years of salvage surgery. The ROC curve analysis showed that the sensitivity of surgical margin status, tumor invasion depth and whether to accept radiotherapy (chemotherapy) after salvage surgery to predict the survival of patients with hypopharyngeal squamous cell carcinoma after 5 years of salvage surgery were 84.15%, 79.60% and 76.43% respectively, and the specificity were 76.03%, 83.51% and 69.46% respectively. The sensitivity and specificity of combined prediction of the three indicators were 92.74% and 77.98% respectively. Conclusion:The overall prognosis of hypopharyngeal squanous cell carcinoma patients after salvage surgery is satisfactory. Positive surgical margin, tumor invasion of muscle, bone tissue or lymph node capsule, and no radiotherapy or chemical therapy after salvage surgery are closely related to poor prognosis. Meanwhile, the combination of surgical margin status, tumor invasion and adjuvant treatment after salvage surgery has good efficacy in predicting postoperative survival benefit of patients.

Significance of neoadjuvant immunotherapy combined with chemotherapy in the treatment of larynx preservation in locally advanced hypopharyngeal squamous cell carcinoma / 临床耳鼻咽喉头颈外科杂志

Objective:To evaluate the clinical significance of neoadjuvant immunotherapy combined with chemotherapy in the treatment of larynx preservation in locally advanced hypopharyngeal squamous cell carcinoma. Methods:Patients with locally advanced HPSCC（cT3-T4aN0-N3M0） were eligible. All received 2 cycles of pembrolizumab combined with docetaxel and platinum neoadjuvant induction therapy. After two cycles, the efficacy was evaluated, followed by radical chemoradiotherapy or surgery and adjuvant chemoradiotherapy according to the efficacy. The primary endpoints were objective response rate（ORR） ，larynx-preservation（LP） rate at 3 months post-treatment and the adverse reactions during neoadjuvant therapy. Results:From December 2021 to December 2022, 10 patients with locally advanced HPSCC（cT3-T4aN0-N3M0） were enrolled. After 2 cycles of the neoadjuvant therapy, 2 patients achieved complete response（CR）, 7 patients achieved partial response（PR）, 1 patient was stable disease（SD）, objective response rate（ORR） was 90%, and disease control rate（DCR） was 100%. 5 patients received radical chemoradiotherapy, 5 patients received surgery and adjuvant chemoradiotherapy, four of them received partial laryngectomy and partial hypopharyngeal resection surgery, and one of them received total laryngectomy and partial hypopharyngeal resection surgery. All patients were able to withstand adverse reactions of neoadjuvant therapy and successfully completed the whole treatment of HPSCC without grade 3-4 treatment-related adverse reactions. There was no recurrence or metastasis during 3-18 months of follow-up. 1 patient died of severe pneumonia 3 months after the completion of radical chemoradiotherapy. At 3 months after treatment, the larynx-preservation rate was 80%. Conclusion:Neoadjuvant immunotherapy combined with chemotherapy has good short-term efficacy and the adverse reactions were tolerable. It can improve the larynx-preservation rate of patients with locally advanced HPSCC, thus improving the prognosis and quality of life of patients.

Comparison of different laryngeal preservation strategies based on chemoradiotherapy in locally advanced hypopharyngeal carcinoma / 临床耳鼻咽喉头颈外科杂志

Objective:This study aimed to evaluate the clinical features and treatment outcomes of the value of response-adapted treatment following radiotherapy and induction chemotherapy follwing subsequent comprehensive therapy in patients with resectable locally advanced hypopharyngeal carcinoma. Methods:This cohort study was conducted from September 2010 to September 2020 in our hospital, 231 patients pathologically confirmed stage Ⅲ and ⅣB resectable locally advanced hypopharyngeal carcinoma included. For the IC-directed ART strategy, IC is used to select good candidates to receive radical RT or CCRT, and others undergo surgery. He response-adapted strategy was determined based on the primary tumor response, which was evaluated at a dose of 50 Gy. If the response reached complete response or partial response（more than 80% tumor regression）, patients received radical RT or CCRT; otherwise, they received surgery, if possible, at 4 to 6 weeks after RT. The end points of the study were OS（overall survival）, progression free survival（PFS）, locoregional recurrence-free survival（LRRFS） and LDFS. Results:In IC-directed group, 75.0%（57/76） patients reached PR after 2 cycles of induction chemotherapy. While in RT-directed group, 70.3%（109/155） patients reached large PR at dose of 50 Gy. The median interquartile range follow-up period of the whole cohort was 63.8 months. The 5-year OS, PFS, LRRFS and SFL of the whole cohort were 47.9%、39.6%、44.3% and 36.2%, respectively. In evaluations based on the different treatment strategies, the 5-year OS and SFL were 51.3% versus 37.0%（HR 0.67; 95%CI 0.43-1.05; P=0.07） and 27.8% versus 39.8%（HR 0.68; 95%CI 0.46-0.99; P=0.04） between IC-directed and RT-directed groups. In additional, surgery complications did not significantly differ between these two groups. Conclusion:In this cohort study, the response-adapted strategy based on an early RT response facilitated better treatment tailoring, and higher laryngeal preservation compared with IC-directed strategies. This approach could provide a feasible laryngeal preservation strategy in patients with resectable locally advanced hypopharyngeal carcinoma.

Analysis of efficacy and prognosis of neoadjuvant chemotherapy and （or） surgery plus radiotherapy for hypopharyngeal squamous cell carcinoma / 临床耳鼻咽喉头颈外科杂志

Objective:To analyze the risk factors that affect the prognosis of patients with hypopharyngeal squamous cell carcinoma（HPSCC） and to compare the efficacy of surgical resection followed by adjuvant radiotherapy（SR） with that of neoadjuvant therapy consisting of platinum-based chemotherapy and fluorouracil combined with either cetuximab or nimotuzumab, followed by SR. The study also aimed to evaluate the overall survival（OS） of patients, their postoperative eating function, tracheostomy decannulation rate, and tumor response to the two neoadjuvant chemotherapies. Methods:A retrospective analysis was performed on the medical records of HPSCC patients who received SR or neoadjuvant therapy followed by SR treatment at the Shanghai General Hospital from 2012 to 2019 and had not undergone any prior treatment. The prognostic factors were analyzed, and the survival analysis of patients who underwent SR treatment with two neoadjuvant chemotherapy regimens was performed. Results:A total of 108 patients were included in the study. The results of the univariate analysis showed that gender（P=0.850） had no significant correlation with the survival rate of HPSCC patients who underwent SR. However, age, smoking history, alcohol consumption history, platelet-to-lymphocyte ratio（PLR）, neutrophil-to-lymphocyte ratio（NLR）, T stage, N stage, neoadjuvant therapy with either cetuximab or nimotuzumab combined with platinum-based chemotherapy and fluorouracil, and histological grade were significantly associated with prognosis（P<0.05）. The multivariate analysis revealed that smoking history, histological grade, and neoadjuvant therapy with either cetuximab or nimotuzumab combined with platinum-based chemotherapy and fluorouracil were independent risk factors affecting the prognosis of HPSCC（P<0.05）. Patients who received neoadjuvant therapy had longer OS than those who underwent SR only（P<0.001）. There was no significant difference in tumor response to the two neoadjuvant therapies and in OS（P>0.05）, and there was no significant difference in the rate of oral feeding and tracheostomy decannulation among the three treatment groups（P>0.05）. Conclusion:Univariate analysis showed that age at tumor onset, smoking history, alcohol consumption history, NLR, PLR, T stage, N stage, whether receiving neoadjuvant chemotherapy, and pathological grade were associated with the prognosis of HPSCC patients receiving SR treatment. Multivariate analysis showed that smoking history, pathological grade, and neoadjuvant chemotherapy were independent risk factors affecting the prognosis. Neoadjuvant chemotherapy with cetuximab or nimotuzumab can prolong the OS of patients, providing a certain basis and reference for the treatment of HPSCC.

Progress on Multi-disciplinary Combined Therapy for Locally Advanced Hypopharyngeal Carcinoma / 肿瘤防治研究

Given the important position and function of the hypopharynx, the preservation of organ function and survival improvement are equally important. The optimal role of multi-disciplinary combined treatment modality becoming increasingly important. The optimal laryngeal preservation strategy for hypopharyngeal cancer is under continuous exploration. With the constant research of surgery, radiotherapy, chemotherapy, and molecular targeted therapy, new laryngeal preservation strategies continue to emerge. Herein, we primarily summarize the advances in multi-disciplinary combined treatment and the future direction in the treatment of hypopharyngeal carcinoma.

The modified Valsalva maneuver in hypopharynx CT scan / 临床耳鼻咽喉头颈外科杂志

Objective:To analyze the significance and factors influencing of CT scan under the modified Valsalva maneuver. Methods:Clinical data of 52 patients with hypopharyngeal carcinoma diagnosed from August 2021 to December 2022 were collected, all patients had calm breathing CT scan and modified Valsalva maneuver CT scan. Compare the exposure effect of the aryepiglottic fold, interarytenoid fold, postcricoid area, piriform fossa apex, posterior hypopharyngeal wall, and glottis with each CT scanning method. The effects of age, neck circumference, neck length, BMI, tumor site, and T stage on the exposure effect were analyzed. Results:In 52 patients, 50 patients（96.15%） completed CT scan at once time. The exposure effect of the CT scan under modified Valsalva maneuver in the aryepiglottic fold, interarytenoid fold, postcricoid area, piriform fossa apex, posterior hypopharyngeal wall was significantly better than CT scan under calm breathing（Z=-4.002, -8.026, -8.349, -7.781, -8.608, all P<0.01）, while CT scan under modified Valsalva maneuver was significantly worse in glottis than CT scan under calm breathing（Z=-3.625, P<0.01）. In the modified Valsalva CT scan, age had no obvious effect on the exposure effect. The exposure effect was better with long neck length, smaller neck circumference, smaller BMI and smaller T stage. The exposure of postcricoid carcinoma was better than pyriform sinus carcinoma and posterior hypopharyngeal wall carcinoma. But differences were not all statistically significant. Conclusion:The anatomical structure of the hypopharynx was clearly under CT scan with modified Valsalva maneuver, which clinical application is simple, but the effect of glottis was worse. The influence of age, neck circumference, neck length, BMI, and tumor T stage on the exposure effect still needs further investigation.

Advances in non-operative treatment of advanced hypopharyngeal squamous cell carcinoma / 国际肿瘤学杂志

Hypopharyngeal squamous cell carcinoma has a high degree of malignancy, hidden location, atypical early symptoms, most patients have reached advanced stage at the time of treatment. Surgical treatment will cause patients to lose laryngeal function and poor postoperative quality of life. The combined application of non-operative therapies such as radiotherapy and chemotherapy can improve the larynx preservation rate of patients and obtain a survival rate similar to that of surgical treatment. Targeted therapy has achieved better results in the treatment of hypopharyngeal squamous cell carcinoma, and immunotherapy has also made great progress in the treatment of head and neck squamous cell carcinoma, which brings new hope for patients with hypopharyngeal squamous cell carcinoma.

Endoscopic submucosal dissection for early hypopharyngeal carcinoma and precancerous lesions / 中华消化内镜杂志

Objective:To evaluate the efficacy and safety of endoscopic submucosal dissection (ESD) for early hypopharyngeal carcinoma and precancerous lesions.Methods:Clinical data of 41 patients who received ESD for early hypopharyngeal carcinoma and precancerous lesions from August 2013 to August 2019 in the Department of Endoscopy of Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College were retrospectively analyzed. Main outcome measurements included operation completion rate, operation time, en bloc resection rate, R0 resection rate, complication rate and recurrence.Results:ESD was successfully completed in all 41 cases, with a success rate of 100.0% and a mean time of 49.1 min (ranged 10-110 min). Fifty-four lesions underwent en bloc resection, with an en bloc resection rate of 98.2% (54/55), of which 41 had negative horizontal and vertical margins, and the R0 resection rate was 74.5% (41/55). During the operation of 55 lesions, there was a small amount of blood oozing on the wound surface, and electrocoagulation with thermal biopsy forceps could successfully stop the bleeding. No perforation occurred, and 2 cases (4.3%) had delayed bleeding after ESD, and hemostasis was successful under emergency endoscopy. Postoperative endoscopy showed that 1 case (2.2%) had esophageal entrance stenosis, and the obstruction was relieved after repeated water balloon dilatation. The follow-up period ranged from 3 to 72 months, and the median time was 18 months. One case was found to have mucosal lesions in the same part of the hypopharynx and received ESD treatment again. Follow-up to October 2020, no residual lesions and recurrence were found.Conclusion:ESD is a safe and effective option for the treatment of early hypopharyngeal carcinoma and precancerous lesions, which is worthy of clinical application.

Progress of microRNA in hypopharyngeal squamous cell carcinoma / 肿瘤研究与临床

microRNA (miRNA) is a class of non-coding single-stranded RNA molecules of approximately 20-22 nucleotides in length, encoded by endogenous genes, which is involved in post-transcriptional regulation of gene expression in plants and animals. miRNA exists in a double-stranded form and is single-stranded when activated. miRNA also plays its role by forming miRNA-induced silencing complex, and is involved in various processes such as cell growth, differentiation, senescence, apoptosis, autophagy, migration and invasion. Hypopharyngeal carcinoma is a kind of rare malignant neoplasm with unknown etiology and poor prognosis. The early symptoms cannot be detected easily, which is prone to missed diagnosis and misdiagnosis. The main pathological type of hypopharyngeal carcinoma is squamous cell carcinoma accounting for 95% that can easily develop cervical lymph node metastasis. The role of miRNA in hypopharyngeal squamous cell carcinoma is of great importance to the targeted therapy of hypopharyngeal squamous cell carcinoma. This paper reviews the progress of the role of miRNA in hypopharyngeal squamous cell carcinoma.

Failure patterns and outcomes after induction chemotherapy followed by radical radiotherapy in patients with locally advanced hypopharyngeal carcinoma / 中华放射医学与防护杂志

Objective:To retrospectively analyze the failure patterns and outcomes of patients with locally advanced hypopharyngeal squamous cell carcinoma (HPSCC) after undergoing induction chemotherapy (IC) followed by definitive radiotherapy.Methods:For patients with locally advanced HPSCC who were treated with IC and definitive radiotherapy from August 2008 to December 2019, their data were collected from the medical records system, and their clinical characteristics, failure patterns, and survival were retrospectively analyzed.Results:A total of 116 eligible patient with squamous cell carcinoma were included in this study. with a median age of 59 (39-79), and 3, 3, 60, and 50 of them had stage Ⅱ, Ⅲ, Ⅳ A, and Ⅳ B HPSCC, respectively. Among these patients, 81 received 1~2 cycles of IC, and 35 received 3-4 cycles of IC. After treatment with IC, 54, 13, and 49 patients received concurrent chemoradiotherapy, radiotherapy combined with targeted therapy, and radiotherapy alone, respectively. The median follow-up was 34.6 months (95% CI: 28.7-40.5 months). The 3-year local recurrence-free survival (LRFS), regional recurrence-free survival (RRFS), metastasis-free survival (MFS), progression-free survival (PFS), and overall survival (OS) of all the patients were 63.5%, 82.8%, 75.2%, 47.3%, and 43.1%, respectively. Median PFS and OS were 26.1 and 28.0 months, respectively. Treatment failure was reported in 59 patients, of whom 22, 5, 12, 10, 3, 6 and 1 experienced local, regional, distant only, local-regional, regional-distant, local-distant, and local-regional-distant failure, respectively. The objective response rate (CR+ PR) of patients after IC was 55.2% (64/116). The LRFS, RRFS, PFS, and OS of IC responders (CR+ PR) were better than those of IC non-responders (SD+ PD) ( χ2 = 12.52, 5.16, 13.19, 11.72, all P< 0.05). Conclusions:IC combined with radical radiotherapy has efficacy to a certain extent in the treatment of locally advanced HPSCC, and locoregional recurrence predominates the failure patterns. The prognosis of IC responders is significantly better than that of IC non-responders.

Time between Diagnosis and Treatment of Hypopharynx and Larynx Cancer: Are Longer Delays Associated with Higher Discrepancy between Clinical and Pathological Staging?

Abstract Introduction At the time of diagnosis, treatment strategies for cancer are largely based upon clinical staging. However, discrepancy between clinical and pathological staging has been reported. Objective To assess the rate of staging discrepancy in Laryngeal and Hypopharyngeal Squamous Cell Carcinoma (LHSCC), the potential influence of higher interval of time from diagnosis to primary surgical treatment, and whether this has any impact on survival outcomes. Methods Retrospective study of patients with LHSCC proposed for primary surgical treatment. Results The study population included 125 Caucasian patients with LHSCC. The level of agreement between clinical and pathological tumor staging was moderate (Cohen's Kappa: 0.400; p < 0.001) and similar result was found for node staging (Cohen' Kappa: 0.520; p < 0.001). The mean time between diagnosis and surgical treatment was 26.66 days and no statistically significant influence was found with staging discrepancy. The sample presented a 5-year Overall Survival (OS) of 58.2% and a Disease-specific survival (DSS) of 72.6%. No statistically significant impact of staging discrepancy on survival was found. Conclusion For advanced LHSCC, based on the findings of physical examination, endoscopy and imaging, is possible to achieve a moderate accuracy between clinical and pathological staging which allows a reliable counselling and treatment planning. Interval of time under 3-4 weeks between diagnosis and surgical treatment does not influence the rate of discrepancy. However, almost 30% of staging discrepancy is expected due to false negatives of imaging and limitations of physical exams.

TRIM21 Promotes Differentiation of Hypopharyngeal Squamous Cell Carcinoma Through Improving Protein Stability by Ubiquitination of Cytoskeleton / 中国生物化学与分子生物学报

E3 ubiquitin ligase TRIM21(tripartite motif containing 21) plays an important role in regulating cell biological functions and clinical prognosis as oncogene or tumor suppressor in different types of tumors. However,the biological functions and molecular mechanism of TRIM21 in hypopharyngeal squamous cell carcinoma (HPSCC) are still unclear.Our results showed that TRIM21 is highly expressed in moderately and well differentiated HPSCC, suggesting the role of TRIM21 in tumor differentiation. Overexpression and knockdown of TRIM21 inhibited or promoted cell proliferation and migration. Meanwhile, the expression of differentiation markers including KRT10 (keratin 10), IVL (involucrin) and TGM1 (transglutaminase 1) were increased and decreased upon TRIM21 overexpression or knockdown, respectively. The bioinformatics analysis of TRIM21 interacting protein identified by co-immunoprecipitation combined with mass spectrometry suggested that TRIM21 may be closely related to the regulation of cytoskeleton protein. We further demonstrated that TRIM21 interacted with KRT10. The inhibition of protein synthesis by cycloheximide led to upregulation of KRT10 in TRIM21 overexpressing FaDu cells, and ectopic expression of TRIM21 enhanced the ubiquitination level of KRT10. In summary, our results suggest that TRIM21 may promote the HPSCC differentiation by mediating ubiquitination of cytoskeleton proteins to improve the protein stability.

Expression of Twist1 and Twist2 in hypopharyngeal carcinoma and its relationship with clinicopathological features / 中国医师杂志

Objective:To investigate the relationship between the expression of Twist1 and Twist2 protein and with clinicopathological features in hypopharyngeal carcinoma.Methods:The clinical and pathological data of 58 patients with hypopharyngeal carcinoma who underwent surgery in Liaoyang Central Hospital from May 2014 to September 2019 were retrospectively analyzed. The general demographic and pathological data of the patients were collected, and the expression of Twist1 and Twist2 protein in the hypopharyngeal cancer tissues and adjacent tissues were detected by immunohistochemistry. The patients were followed up to record their survival. The relationship between protein expression of Twist1 and Twist2 and clinical pathological stages and prognosis was analyzed.Results:Immunohistochemistry showed that the positive expression rates of Twist1 and Twist2 protein in 58 patients with hypopharyngeal cancer were significantly higher than those in adjacent tissues ( P<0.05). The positive expression rates of Twist1 and Twist2 in patients with T stage T 3+ T 4 and clinical stage Ⅲ+ Ⅳ were significantly higher than those in patients with T 1+ T 2 and Ⅰ+ Ⅱ ( P<0.05). The positive expression rates of Twist1 and Twist2 in patients with lymph node metastasis were significantly higher than those without lymph node metastasis, and the positive expression rates of Twist1 and Twist2 in low differentiation were significantly higher than those in high differentiation ( P<0.05). 52 patients were followed up postoperatively, the follow-up rate was 89.67%, among which 28 patients recurred (53.85%), and the 1-, 3-, and 5-year survival rates were 80.77%(42/52), 53.85%(28/52), and 21.15%(11/52), respectively. The survival rate of patients with positive expression of Twist1 and Twist2 protein was significantly lower than that of patients with negative expression ( P<0.05). Multiple analysis showed that recurrence, tumor stage, positive expression of Twist1 and Twist2 protein were independent risk factors for survival ( P<0.05). Conclusions:The expression of Twist1 and Twist2 protein in hypopharyngeal carcinoma is stronger than that in normal tissues. The expression of Twist1 and Twist2 protein is related to T stage, clinical stage, pathological tissue type and lymph node metastasis. Meanwhile, the high expression of Twist1 and Twist2 proteins is an independent factor affecting the prognosis of patients. They may be potential indicators to judge the prognosis of patients with hypopharyngeal cancer.

Changing trend in prognosis of primary hypopharyngeal carcinoma and reason analysis: date comparison of 2003-2007 and 2010-2014 in a single centre / 中华耳鼻咽喉头颈外科杂志

To explore changing trend in prognosis of primary hypopharyngeal carcinoma and to analyze the reasons at the Eye, Ear, Nose and Throat Hospital of Fudan University. We retrospectively analyzed the clinical data of 461 patients with primary hypopharyngeal carcinoma treated at the Eye, Ear, Nose and Throat Hospital of Fudan University from 2003 to 2007 (Group 1) and 2010 to 2014 (Group 2) according to the inclusion criteria. 142 from Group 1, including 133 males and 9 females, rangedfrom 38 to 82 years old and 319 from Group 2, including 313 males and 6 females, ranged from 39 to 81 years old, were included in this work. The laryngeal function preservation rate, survival outcome, application and effect of pre-and post-operative adjuvant therapy were compared. SPSS 24.0 was used for statistical analyses. There were 62 patients with early disease (T1-2N0) including 18 in Group 1 and 44 in Group 2, in whom 3 (16.7%) underwent surgical procedures with laryngeal function preservation in Group 1, while, 30 (68.2%) underwent laryngeal function preservation surgery in Group 2. The laryngeal function preservation rate showed an obviously upward trend in recent years (χ(2)=13.617, 0.001), whereas, the recurrence-free survival rate (RFS) and overall survival rate (OS) showed no significant differences between two groups (0.469 and 0.808, respectively). Among the 399 patients with advanced disease, 124 were in Group 1 and 275 in Group 2. After propensity score matching (PSM) was used, the OS rate was significantly higher for Group 2 than Group 1 (0.017), while the application of laryngeal function preservation surgery was significantly higher in Group 2 (χ(2)=4.686, 0.030). The application rates of preoperative adjuvant therapy and postoperative adjuvant chemotherapy were significantly higher in group 2 than in group 1 (χ(2)=5.687, 0.017; χ(2)=19.407, 0.001). The application of laryngeal function preserving surgery significantly increases the retention rate of laryngeal functions in patients with early-stage hypopharyngeal carcinoma, with similar long-term survival outcomes. The application of comprehensive treatment including preoperative adjuvant therapy and postoperative adjuvant treatments, especially postoperative chemoradiotherapy, shows an obvious increase in the function preservation rate and long-term survival in patients with advanced hypopharyngeal carcinoma.

Analysis for potential targeting genes of TPF regimen induction chemotherapy in hypopharyngeal squamous cell carcinoma / 中华耳鼻咽喉头颈外科杂志

To analyze the differentially expressed genes related to the chemosensitivity with the TPF regimen for hypopharyngeal squamous cell carcinoma and to measure potential functional targeting genes expressions. Twenty-nine patients with primary hypopharyngeal cancer who underwent induction chemotherapy with TPF from January 2013 to December 2017 in Beijing Tongren Hospital were enrolled for microarray analysis, including 28 males and 1 female, aged from 43 to 73 years old. Among them, 16 patients were sensitive to chemotherapy while 13 patients were non-sensitive. Illumina Human HT-12 Bead Chip was applied to analyze the gene expressions and online bioinformatics analysis was used to analyze the differentially expressed genes. Reverse transcription and quantitative real-time PCR (RT-qPCR) was used to measure the mRNA expression of potential functional genes of TPF induction chemotherapy in 43 samples, 29 from original patients and 14 from additional patients. Graphpad prism 7.0 software was used for statistical analysis. A total of 1 381 significantly differentially expressed genes were screened out. By GO analysis, up-regulated genes included sequestering in extracellular matrix, chemokine receptor binding and potassium channel regulator activity; down-regulated genes included regulation of angiogenesis, calcium ion binding and natural killer cell activation involved in immune response. With KEGG database analysis, down-regulated pathways included ECM-receptor interaction and peroxisome and up-regulated pathways included Glutathione metabolism and PPAR signaling pathway. The expressions of CD44 and IL-6R were significantly different and appeared biologically significant. CD44 was significantly upregulated in insensitive tissues (0.54±0.06) compared with sensitive tissues (0.33±0.04)(0.01). IL-6R was significantly downregulated in insensitive tissues (0.44±0.03) compared with sensitive tissues. (0.68±0.03) (0.01). CD44 and IL-6R may be potentially functional genes of TPF induction chemotherapy in hypopharyngeal squamous cell carcinoma.