Prognostic Significance of Epidermal Growth Factor Receptor Phosphorylation and Mutation in Head and Neck Squamous Cell Carcinoma

The molecular status of the epidermal growth factor receptor (EGFR) has not been as well studied in head and neck squamous cell carcinoma (HNSCC) as in lung cancer. We examined the frequencies of EGFR mutations as well as the expression/phosphorylation status of the EGFR protein in HNSCC patients. Moreover, we tried to elucidate associations between EGFR molecular status and patient characteristics and disease‐free survival. In this prospective cohort study, clinical data and samples were obtained from 82 consecutive patients who had not been treated with EGFR molecular targeting therapy. Full‐length EGFR was sequenced, and expression and phosphorylation of the EGFR protein were measured by Western blotting. Four novel mutations (E709K, V765G, Ins770G, and G1022S) and one mutation well‐known in lung cancer (L858R) were identified in six HNSCC samples (7%), but we could not find any mutations in the extracellular domain of EGFR, such as EGFRvIII, in this study. E709K and Ins770G as well as L858R appear to be functional mutations based on the use of Ba/F3 cells. In terms of patient characteristics, the number of metastatic lymph nodes and node stage were associated with phosphorylation of EGFR. No patients with EGFR mutations relapsed during the study period. Excluding mutated cases, patients whose tumor samples showed phosphorylated EGFR relapsed significantly earlier than those without phosphorylated EGFR. This finding was still significant after adjusting for mutation and overexpression of EGFR protein using the Cox proportional hazard model. In conclusion, phosphorylated EGFR without mutations may be a marker of poor prognosis in patients with HNSCC.     

Overexpression of BASP1 Indicates a Poor Prognosis in Head and Neck Squamous Cell Carcinoma

Objective: Brain abundant membrane attached signal protein 1 (BASP1) was originally identified as a membrane and cytoplasmic protein. Recent studies have shown that BASP1 highly expressed in cancer and promoted the proliferation of cancer. However, the role of BASP1 in head and neck squamous cell carcinoma (HNSCC) is largely unknown.  Here, we performed a systematic data analysis to examine whether BASP1 can function as prognostic marker in HNSCC. Methods: In this study, we used Oncomine, and UALCAN, databases to analyze the expression of BASP1 in HNSCC. We used Kaplan-Meier plotter to evaluate the effect of BASP1 on clinical prognosis. In addition, we also analyzed genetic alterations, interaction network, and functional enrichment of BASP1. Results: BASP1 mRNA expression level was remarkably increased in HNSCC than in normal tissues (P=1.624e-12). Moreover, high BASP1 expression was significantly related to poor survival (p=0.00056) in HNSCC patients. In addition, BASP1 gene amplified in 5% of HNSCC patients which contributes to the overexpression of BASP1. Conclusions: These findings suggest that BASP1 was frequently amplified which contributes to the overexpression of BASP1, thereby promoting HNSCC progression. Thus, these results indicate that BASP1 might serve as a biomarker to predict the progression and prognosis of HNSCC patients.     

Protein Expression of Stromelysin-2 in Head and Neck Squamous Cell Carcinomas

Background: Some matrix metalloproteinases (MMPs) are involved in invasion and metastasis of head and neck squamous cell carcinoma (HNSCC). However, there are few studies on association between stromelysin-2 (ST-2) and invasive behavior of HNSCC. The purpose of this study was to investigate Stromelysin-2 expression by immunohistochemistry. Materials and Methods: This study was conducted on 81 specimens, including 61 HNSCC and 20 non neoplastic epithelium. Sections with 5 micron thickness were prepared and stained with immunohistochemistry technique. Then expression of ST-2 was evaluated according to percentage of stained cells and intensity of staining. Data were analyzed by SPSS (V.21) using Kruskal-Wallis and Tukey tests (P<0.05). Results: The 61 HNSCC specimens were grades I 36.1%, II 34.4% and III 29.5%. The level of ST-2 expressions were moderate (++) and intensive (+++) in 21.3% and 78.7% of tumors, respectively. The ST-2 expression level was only significant between the tumors with grade I and grade III (P=0.016). Tumors presented ST-2 expression with staining intensity of mild 6.6%, moderate 26.2% and strong 67.2%. Staining intensity of ST-2 in grade I tumors was significantly lower than grade II and grade III (P<0.05), and there was no significant difference between grades II and III (P=0.99). Conclusions: According to this study, the expression of ST-2 is associated with histopathological grade and tumor differentiation in HNSCCs.     

Tumor Infiltrating Lymphocytes are Prognostic Factors and Can Be Markers of Sensitivity to Chemoradiotherapy in Head and Neck Squamous Cell Carcinoma

Background: Tumor-infiltrating lymphocytes (TILs) are assessed by the ratio of the area of lymphocytes infiltrating the stroma. TILs are important in breast cancer and malignant melanoma and are being established as a marker of prognosis and sensitivity to chemotherapy. This has resulted in various therapies being developed in fields such as breast cancer. However, the evaluation of TILs in head and neck squamous cell carcinoma (HNSCC) is not progressing, and the prognosis is still poor. Thus, investigating whether or not the evaluation of TILs is also effective in HNSCC and prognoses can be predicted with just biopsy samples alone is required. Methods: This study included 153 patients who were diagnosed with HNSCC between January 2010 and December 2019, underwent treatment, and could be followed up thereafter at our institution. Results: TILs, overall survival (OS), and progression-free survival (PFS) were evaluated in all patients, the chemoradiotherapy arm, and the surgery arm. The cut-off value for TILs was 50%. In all patients, OS was 69.8% and 40.2% (P = 0.01) and PFS was 58.4% and 31.6% (P = 0.003) in the high and low TIL groups, respectively. Multivariate analyses revealed that TILs independently predicted prognosis. In the chemoradiotherapy arm, OS was 70.8% and 31.6% (P = 0.012) and PFS was 63.4% and 20.3% (P = 0.001) in the high and low TIL groups, respectively. No significant differences were noted in the surgery arm. Conclusions: In HNSCC, TILs can be used as a prognosis predictor and chemoradiotherapy biomarker. Assessments can be performed just with hematoxylin–eosin staining and is very simple. This will greatly contribute to report personalized therapy progress. Further evaluations and, thus, prospective clinical multicenter trials are needed to use TILs in clinical practice for HNSCC.     

头颈鳞状细胞癌手术切缘P53表达及意义

 目的 探讨头颈鳞癌手术切缘P53蛋白的表达及其与肿瘤临床病理学参数和局部复发的关系。 方法 用免疫组织化学的方法检测82例头颈鳞癌手术切缘标本和10例正常口腔黏 膜组织中P53的表达情况,并分析其与肿瘤局部复发和临床病理参数的关系。 结果 在82例头颈鳞癌患者中,切缘突变型P53阳性表达率为59.76%(49/82)。阳性表达主要定 位于切缘上皮基底细胞核。突变型 P53蛋白在头颈鳞癌手术切缘中的表达显著高于其在正常口腔粘膜组织中的表达(P< 0.05)。头颈鳞癌患者手术切缘突变 型P53的表达与肿瘤分化程度和局部复发显著相关(P<0.05), 但与患者年龄、性别、 肿瘤病理分期、淋巴结转移及肿瘤部位无关(P>0.05)。 结论 切缘突变型P53的阳性表达与头颈鳞癌术后肿瘤的进展有关,它有望成为判定肿瘤局部复发的 预测因子。     

Cancer Stem Cells in Head and Neck Squamous Cell Carcinoma: A Review

Research indicates that a small population of cancer cells is highly tumorigenic, endowed with the capacityfor self-renewal, and has the ability to differentiate into cells that constitute the bulk of tumors. These cells areconsidered the ‘‘drivers’’ of the tumorigenic process in some tumor types, and have been named cancer stemcells (CSC). Epithelial-mesenchymal transition (EMT) appears to be involved in the process leading to theacquisition of stemness by epithelial tumor cells. Through this process, cells acquire an invasive phenotype thatmay contribute to tumor recurrence and metastasis. CSC have been identified in human head and neck squamouscell carcinomas (HNSCC) using markers such as CD133 and CD44 expression, and aldehyde dehydrogenase(ALDH) activity. Head and neck cancer stem cells reside primarily in perivascular niches in the invasive frontswhere endothelial-cell initiated events contribute to their survival and function. Clinically, CSC enrichment hasbeen shown to be enhanced in recurrent disease, treatment failure and metastasis. CSC represent a novel targetof study given their slow growth and innate mechanisms conferring treatment resistance. Further understandingof their unique phenotype may reveal potential molecular targets to improve therapeutic and survival outcomesin patients with HNSCC. Here, we discuss the state-of-the-knowledge on the pathobiology of cancer stem cells,with a focus on the impact of these cells on head and neck tumor progression, metastasis and recurrence due totreatment failure.     

Wnt Signaling: A Potential Therapeutic Target in Head and Neck Squamous Cell Carcinoma

Cellular maintenance and development are two fundamental mechanisms regulated by the canonical Wnt signalingpathway. Wnt/beta-catenin signaling pathway controls a myriad of cellular processes that are essential for normal cellfunctioning. Cell cycle progression, differentiation, fate determination, and migration are generally orchestrated bycanonical Wnt signaling. Altered Wnt/beta-catenin signaling has been considered a promoting event for different typesof cancers and the oncogenic potential of Wnt signaling have been discussed in many cancer types, including breast,colon, pancreatic as well as head and neck. Furthermore, Wnt signaling is critical for the maintenance and stemnessof both the normal as well as cancer stem cells. This review sheds new light on Wnt signaling and explains how it canregulate normal physiological processes and curtail the development of cancer. It depicts the vital functions of Wntsignaling in the stem cell growth and differentiation by focusing on current druggable targets that have been ascribedby recent studies. Thus, Wnt signaling pathway retains a tremendous potential in eradicating head and neck squamouscell carcinoma.     

Characteristics and Impact of HPV-Associated p16 Expression on Head and Neck Squamous Cell Carcinoma in Thai Patients

Background: Head and neck squamous cell carcinoma (HNSCC) is a common malignancy in Asia. Infection by human papilloma virus (HPV) has been recognized as an etiological risk for HNSCC, especially oropharyngeal region. While the association between HPV and HNSCC has been well evaluated in Western countries, only a few investigated the HPV-associated HNSCC in Southeast Asia. This study evaluated the prevalence, the characteristics, and the impact of HPV on the treatment outcomes in Thai HNSCC patients. Methods: Non-nasopharyngeal HNSCC patients treated at Ramathibodi Hospital during 2007-2013 were identified through the cancer registry database. Baseline patient, treatment data and survivals were retrospectively reviewed. The formalin-fixed paraffin-embedded (FFPE) tissue sections were retrieved for p16 analysis. The HPV status was determined by p16 immunohistochemistry. The survival outcomes were analyzed in cases which p16 status was confirmed. Results: Total of 200 FFPE tissues of HNSCC patients was evaluated for p16 expression. Positive p16 status was observed in 24 cases (12%); majority of p16-positive were men (20:4 cases). The oropharynx (37.9%) was the most common site found in p16-positive while oral cavity (3.2%) was the least common site. Interestingly, 66.7% of p16-positive were former/current smokers, and 70.8% of this subgroup was categorized as clinical AJCC stage III-IV. The p16-positive HNSCC was significantly superior in 5-year overall survival [5-yrs OS 63% vs. 40%, p=0.03], 5-year disease-free survival [5-yrs DFS 61% vs. 36%, p=0.03] and in 5-year locoregional relapse-free survival [5-yrs LRFS 93% vs. 68%, p=0.018] when compared with p16-negative. Conclusions: In comparison to the results from the Western countries, the prevalence of HPV-related HNSCC in Thai patients was less, and differences in some characteristics were observed. Nevertheless, improvement in OS, DFS and LRFS were observed in p16-positive patients. Our analyses suggested that p16 status is also a strong prognostic marker for HNSCC patients in Thailand.     

High p16 Expression Predicts a Positive Response to Chemoradiotherapy in Stage IVa/b Head and Neck Squamous Cell Carcinoma

Background: The objective of this study was to evaluate the effect of p16 expression on response tochemoradiation in stage IVa/b head and neck squamous cell carcinoma (HNSCC) patients. Methods: Weretrospectively identified 64 patients with stage IVa/b HNSCC who received chemoradiation. Eligibility criteriaincluded presence of biopsy-proven stage IVa/b HNSCC without a prior history of chemotherapy or radiotherapy.Immunohistochemistry was used to assess p16 protein expression in pretreatment biopsy specimens. Results: Ofthe 64 patients, 38 showed high p16 expression, and 50 patients responded to chemoradiotherapy, 32 exhibitinga complete and 18 a partial response. Response was significantly associated with p16 expression (P<0.001)and multivariate analysis indicated that that p16 expression (HR: 2.62, 95%C.I.: 1.14-6.06; P=0.024) was anindependent prognostic factor for overall survival. Conclusions: High p16 expression predicts a better responseto chemoradiation in patients with stage IVa/b HNSCC.     

Serum Vascular Endothelial Growth Factor-A (VEGF-A) as a Biomarker in Squamous Cell Carcinoma of Head and Neck Patients Undergoing Chemoradiotherapy

Background: To evaluate serum VEGF-A levels in squamous cell carcinoma of head and neck (SCCHN)patients and relationships with response to therapy. Materials and Methods: Serum VEGF-A levels in patients(n=72) treated with radiotherapy (RT) or radio-chemotherapy (RCT) and controls (n=40) were measured byELISA. Results: Serum VEGF-A levels of the SCCHN cases were significantly higher (p=0.001) than in healthycontrols, and in patients with positive as compared to negative lymph node status (p=0.004). Similarly, patientswith advanced stage (Stage III-IV) disease had more greatly elevated levels of serum VEGF-A level than theirearly stage (Stage I-II) counterparts (p=0.001). In contrast, there was no significant difference (p=0.57) inserum level of VEGF-A in patients with advanced T-stage (T3-4) as compared to early stage (T1-2). Similarly,patients with distant metastasis had no significant (p=0.067) elevation in serum VEGF-A level as compared tonon-metastatic disease. However, the non-responder patients had significantly higher serum VEGF-A level ascompared to responders (p=0.001). Conclusions: Our results suggest that the serum VEGF-A level may be auseful biomarker for the prediction of response to therapy in SCCHN.     

乙酰肝素酶和VEGF在头颈鳞癌组织中的表达及其临床意义

目的: 探讨HPSE、VEGF蛋白在头颈鳞癌组织中的表达及其临床意义,并探讨其相关性。 方法: 采用免疫组织化学(S-P法)和流式细胞检测方法对62例头颈鳞癌组织中HPSE和VEGF蛋白的表达进行检测,并采用计算机图像分析进行定量分析。 结果: HPSE和VEGF蛋白在头颈鳞癌组织中的表达均高于正常粘膜组织(P<0.01);淋巴结转移组鳞癌组织中HPSE和VEGF的表达高于无淋巴结转移组(分别为P<0.05和P<0.01);淋巴结转移组中原发灶与转移灶中HPSE和VEGF的表达差异无统计学意义(P值均>0.05),但原发灶与转移灶之间的表达成正相关(分别r=0.9162,r=0.9278,P均<0.01);HPSE和VEGF的表达均与头颈鳞癌组织中TNM分期有关(P均<0.05);HPSE与VEGF在头颈鳞癌组织中的表达呈正相关关系(r=0.7602,P<0.01);HPSE和VEGF的表达分别与患者的生存期存在负相关关系(分别为r=-0.647,r=-0.568,P均<0.01)。 结论: 头颈鳞癌组织中HPSE和VEGF蛋白的表达与TNM分期、淋巴结转移和预后有密切关系,两种基因起头颈鳞癌发生、发展中起重要作用。     

Role of Tobacco in the Development of Head and Neck Squamous Cell Carcinoma in an Eastern Indian Population

Head and neck squamous cell carcinoma (HNSCC) accounts for about 30-40% of all cancer types in Indiaand the subcontinent in general. HNSCCs are primarily not hereditary, but rather a disease of older and middleaged adults. Many etiological factors like tobacco, alcohol and HPV infection are known to play importantroles. Eastern India, particularly Kolkata, has a population heavily exposed to various types of smoked andsmokeless tobacco, with only limited exposure to alcoholic beverages. Since there have been no previousepidemiological studies on tobacco as the main risk factor for head and neck carcinogenesis in Kolkata, we herecarried out a hospital based case control study in the city and its adjoin regions. Data from 110 patients diagnosedwith HNSCC and a similar number of matched control samples were analyzed using chi-square (χ2) test. Survivalstatus of the patients was also analyzed using the Kaplan-Meier method. A tobacco habit was significantlycorrelated with the incidence of HNSCC and persons with current addiction had a 2.17 fold increased risk ofcancer development. Dose-response relationships were seen for the frequency (p=0.01) and duration (p=0.02) oftobacco exposure with the risk. No significant difference in impact was found with smoked as opposed to smokelesstobacco in the development of the disease. Among HNSCC patients, significant poor survival in cases withtobacco habit than in those with no addiction and in cases with >10 years of addiction than in those with ≤ 10years of addiction. Our data suggest that tobacco in both smoked and smokeless forms is the most importantrisk factor for both development and prognosis of HNSCCs and may be a major source of field cancerization onthe head and neck epithelium in the eastern Indian population.     

Expression of Tumor-associated Glycoantigen, Sialyl Lewisa, in Human Head and Neck Squamous Cell Carcinoma and Its Application to Tumor Immunotherapy

The glycoantigen sialyl Lewis^a (sLe^a) is widely expressed on a variety of gastrointestinal tumor cells. Here, we immunohistochemically demonstrated the expression of sLe^a antigen in 54% (7 out of 13) of human head and neck squamous cell carcinoma (H-NSCG) samples. Frequent expression of sLe^a antigen was also demonstrated on a variety of H-NSCC cell lines using flow cytometry. Both CD4⁺ and CD8⁺ T cells, which were activated with immobilized OKT3 monoclonal antibody plus inter-leukin-2, showed augmented cytotoxicity against sLe^a-positive H-NSCC, including autologous tumor cells, on targeting with anti-CD3 x anti-sLe^a bispccific antibody, suggesting that sLe^a antigen is a good target molecule for bispecific antibody-dependent adoptive tumor immunotherapy of human head and neck cancer.     

头颈部鳞癌肿瘤间质液压的检测

目的：为了检测头颈部鳞癌病人的肿瘤间质液压（TIFP）。方法：采用国产ZYH－3型组织液压测量仪，检测了16例头颈部鳞癌原发灶及4例转移灶的TIFP。结果：发现头颈部鳞癌原发灶和转移灶的TFFP都增高，测得的TIFP值与肿瘤体积具有良好的相关关系。结论：TIFP增高是恶性肿瘤的一项重要病理生理学特征。TIFP增高是抗癌药和免疫因子进入肿瘤的病理学障碍，因此，设法降低TIFP具有重要意义。     

头颈鳞癌的术前化疗效果

[目的]研究术前化疗能否提高手术切除率,降低局部复发率。［方法］104例头颈鳞癌病人分成试验组（58例）：术前化疗 手术 术后放疗组;对照组（46例）：手术 术后放疗,但不用术前化疗。手术方式为原发癌切除 颈淋巴结清扫术,用或不用肌皮瓣修复头颈部组织缺损。术后放疗剂量50Gy～60Gy。术前化疗用PFP方案即DDP＋5－Fu＋PYM。［结果］试验组部分缓解67.2%,微效17.2%。试验组和对照组3年局部复发率分别为27.6%、52．2％（P＜0.01）;但3年内远处转移率无差异,分别为34．5%、34．8%,中位生存期分别为23个月和22个月,3年生存率分别为65．5%和56．5%（P＞0.05）。［结论］术前新辅助化疗,可以缩小瘤体,提高手术切除率,减少局部复发。     

CD 44V 3 在甲状腺癌中的表达及临床意义研究

 目的　探讨转移相关基因 CD 44V3在甲状腺癌中的表达及其临床意义。方法　用免疫组织化学 S- P法 ,检测 40例甲状腺癌、2 0例甲状腺腺瘤和 1 0例正常甲状腺组织中 CD44V3基因蛋白的表达情况 ,并分析其与相关临床病理因素的关系。结果　CD44V3表达阳性率在甲状腺癌组显著高于腺瘤组和正常甲状腺组。有转移者 CD44V3阳性率高于无转移者 ,转移灶阳性率高于原发灶。CD44V3阳性表达与肿瘤大小、病理组织类型无关。结论　CD44V3与肿瘤浸润和转移相关 ,可作为预测肿瘤转移潜能的指标。     

口腔鳞状细胞癌中MMP9、CD44v6表达及其意义

[目的]研究基质金属蛋白酶-9（matrix metalloproteinase-9,MMP9）、CD44拼接变异体6（CD44 splice variant 6,CD44v6）在口腔鳞状细胞癌（OSCC）中的表达及其相互关系.[方法]应用S-P免疫组化法检测82例口腔鳞状细胞癌,40例口腔黏膜鳞状上皮非典型增生和30例正常口腔黏膜中MMP9和CD44v6的表达.[结果]（1）MMP9在OSCC组中的阳性率（84.1%）显著高于非典型增生组（17.5%）和正常口腔黏膜组（6.6%）.MMP9的阳性表达率与淋巴结转移、肿瘤分化程度相关.（2）CD44v6在OSCC组中的阳性率（78.4%）显著高于非典型增生组（30.0%）和正常口腔黏膜（13.3%）.CD44v6的高表达率与淋巴结转移相关.（3）OSCC中MMP9的表达与CD44v6表达呈正相关.[结论]MMP9、CD44v6对OSCC发生过程具有重要意义,MMP9和CD44v6的表达在OSCC淋巴结转移过程中可能呈协同作用.