口腔鳞癌中HPV16、18型感染和p53蛋白表达的检测研究

目的探讨人类乳头状瘤病毒(human papillomavirus, HPV)感染、p53蛋白表达在口腔鳞状细胞癌发生、发展中的作用与相互关系.方法采用聚合酶链反应(PCR)检测HPV16、18型DNA;采用免疫组织化学LSAB法检测p53蛋白产物在细胞核中的表达.结果口腔鳞癌组中HPV16、18 DNA总阳性率为48.89%(22/45),p53蛋白表达阳性率为62.22%(28/45).HPV16、18型感染组及p53蛋白过度表达组的平均生存期、生存率均低于无HPV感染组(P>0.05)及无p53过度表达组(P<0.05).结论 HPV16、18型感染、p53基因突变与口腔鳞癌的发生密切相关.口腔鳞癌患者的预后在一定程度上与HPV感染、p53表达状况有关.     

EGFR and Ki‐67 expression in oral squamous cell carcinoma using tissue microarray technology

J Oral Pathol Med (2010) 39 : 571–578 Objective: Our aim was to validate the use of tissue microarrays (TMA) in oral squamous cell carcinomas (OSCC) to analyse epidermal growth factor receptor (EGFR) and Ki‐67 expression. We also analysed the relationship that the expression of these markers may have with clinical, pathological and survival variables. Patients and methods: The study sample comprised 39 unselected patients diagnosed and treated for OSCC. We analysed Ki‐67 and EGFR expression by immunohistochemistry on formalin‐fixed, paraffin‐embedded surgical specimens. Whole sections (WS) were compared with double 1.5 mm core‐tissue microarrays. Results: High EGFR expression was observed both on TMA (in 98% of the cases) and WS (in 100% of the cases) with substantial agreement kappa value (0.720). EGFR expression was not significantly associated with clinical, pathological and survival variables on TMA and WS. Ki‐67 analysis showed a Spearman correlation of 0.741 with a Ki‐67 mean labelling index of 45% in TMA and 56.8% in WS. We found a significant relationship between gender and Ki‐67 labelling index on WS (P = 0.022) and TMA (P = 0.002). Clinical stage was the only parameter in multivariate analysis that had a significant predictive value. Conclusion: We demonstrate that dual 1.5 mm core TMA is a valid, rapid, economical and tissue‐saving way to study OSCC biopsies and that it presents strong correlation with the WS. EGFR overexpression in OSCC suggests that these tumours may be a candidate for therapy investigation directed to EGFR.     

Nuclear fractal dimension in oral squamous cell carcinoma: a novel method for the evaluation of grading,staging, and survival

Fractal dimension (FD) in tissue specimens from patients with oral squamous cell carcinoma (OSCC) was evaluated. FD values in different stages of OSCC, and the correlations with clinicopathological variables and patient survival were investigated. Histological sections from OSCC and control non‐neoplastic mucosa specimens were stained with hematoxylin–eosin for pathological analysis and with Feulgen for nuclear evaluation. FD in OSCC groups vs. controls revealed statistically significant differences (P < 0.001). In addition, a progressive increase of FD from stage I and II lesions and stage III and IV lesions was observed, with statistically significant differences (P = 0.003). Moreover, different degrees of tumor differentiation showed a significant difference in the average nuclear FD values (P = 0.001). A relationship between FD and patients' survival was also detected with lower FD values associated to longer survival time and higher FD values with shorter survival time (P = 0.034). These data showed that FD significantly increased during OSCC progression. Thus, FD could represent a novel prognostic tool for OSCC, as FD values significantly correlated with patient survival. Fractal geometry could give insights into tumor morphology and could become an useful tool for analyzing irregular tumor growth patterns.     

经首次根治性手术治疗口腔鳞状细胞癌患者的生存相关影响因素分析

目的分析口腔鳞状细胞癌(OSCC)患者的总体生存率,以及影响生存率的临床病理因素。方法采集对首次接受根治性外科手术治疗的78例OSCC患者的临床病理及随访资料进行回顾性分析。对计数、计量资料进行描述性分析;采用Kaplan-Meier法绘制生存曲线;采用COX比例风险回归模型进行单因素和多因素分析,分析患者的生存率及预后相关影响因素。结果最终纳入生存分析的患者共计68例,中位随访时间为63(6~87)个月,5年总体存活率为55.9%,随访期间因OSCC死亡患者的中位生存时间为20.5(6~52)个月。单因素分析表明,临床分期、原发灶大小、淋巴结转移、病理分化及复发转移是影响生存时间的暴露因素(P<0.05);多因素分析表明,病理分化、复发转移是影响生存时间的独立危险因素(P<0.05)。78例OSCC患者中合并发生食道鳞状细胞癌(ESCC)者有4例(5.1%)。结论根据肿瘤的临床分期(TNM分期)、原发灶大小、淋巴结转移、病理分化及复发转移可对患者的生存预后作出一定的预测,其中病理分化及复发转移是影响生存预后的独立危险因素。有吸烟饮酒史的OSCC患者应常规进行ESCC临床筛查。     

Risk of oral cancer associated with human papillomavirus infection, betel quid chewing, and cigarette smoking in Taiwan — an integrated molecular and epidemiological study of 58 cases

BACKGROUND: The association between oral squamous cell carcinoma (OSCC) and human papillomavirus (HPV) 6, 11, 16 and 18 is uncertain. Past reports varied in the methodology and results. We conducted this study using in situ PCR in situ hybridization (ISH) assay which was considered as the most sensitive method for detection of viral DNA. We undertook an epidemiologic survey about the history of betel quid chewing and cigarette smoking, since these habits are common in Taiwan. METHODS: In situ PCR ISH was performed on the tumor specimens from 29 patients with OSCC and the oral mucosal specimens from 29 patients without OSCC. Their betel quid chewing and cigarette smoking histories were also reviewed. RESULTS: HPV16, HPV18, betel quid chewing and cigarette smoking were statistically significant risk factors in univariate analysis. HPV6 and 11 were not. Multivariate analysis showed that HPV16 infection (adjusted Odds ratio = 11.20) and betel quid chewing (adjusted Odds ratio = 17.06) remained to be independent factors for OSCC. CONCLUSIONS: Our results showed that HPV16 and betel quid chewing were two major risk factors for OSCC in Taiwan, indicating that they act through different mechanisms in the pathogenesis of OSCC.     

Detection of mutation-specific epidermal growth factor receptor (E746–A750del) and lack of detection of human papillomavirus in oral squamous cell carcinoma

The clinical impact of epidermal growth factor receptor (EGFR) (E746–A750del) mutation and human papillomavirus (HPV) in oral squamous cell carcinoma (OSCC) is unclear. EGFR (E746–A750del) expression was analyzed in OSCC specimens (n = 161) by immunohistochemistry. The expression results were correlated with clinical characteristics and impact on survival. Using INNO-LiPA Extra, high-risk HPV types were genotyped and analyzed in 211 OSCC specimens. Positive EGFR (E746–A750del) expression (n = 40/161, 25%) was not associated with any clinicopathological characteristics, prognostic factors, social habits (smoking, alcohol consumption), or tumour-specific survival. HPV16 DNA was detected in three out of 211 samples (HPV16-positive: n = 3/211, 1.4%). This study shows that mutation-specific EGFR (E746–A750del) expression and HPV do not appear to be relevant to the survival of patients with OSCC.     

Expression of CA-IX is associated with advanced stage tumors and poor survival in oral squamous cell carcinoma patients

J Oral Pathol Med (2012) Introduction: Carbonic anhydrases (CAs), a group of ubiquitously expressed metalloenzymes, are involved in numerous physiological and pathological processes, including tumorigenicity. Specifically, CA‐IX has been primarily found in hypoxic tumor tissues. Material and methods: This is a retrospective study of tumors from the Tissue Bank of the Pathology Department of the University Hospital of Santiago de Compostela. We selected 50 oral squamous cell carcinomas (OSCCs) using Tissue Microarray (TMA) technology. The immunohistochemical study was performed to determine CA‐IX expression. The resulting data were subject to statistical analysis and survival curves. Results: Of the 50 cases, 23 were detected in early stages (I and II) and 27 in advanced stages (III and IV). In the first year, almost 50% of patients in stages III–IV died, which contrasted with those patients in initial stages who registered a survival rate of 80% (P = 0.019). Regarding the expression of CA‐IX, nine cases (18%) were negative, 18 cases (36%) were moderate, while 23 cases (46%) were intense. Tumors in stages I‐II showed a positivity of 52.6%; however, in advanced stages, the percentage reached 95.5% (P = 0.002). Regarding CA‐IX expression and survival, patients with tumors with strong staining had a lower average survival time (13.8 months) than patients with negative or weak‐moderate staining (33.4 and 32.8 months, respectively), log‐rank = 6.1, P value = 0.0484. Conclusions: Early diagnosis of these tumors is essential to improve patient survival. CA‐IX expression augments with increasing tumor stage, probably related with the degree of hypoxia; thus, its measurement can be used as a prognostic factor.     

Human papillomavirus type 16 and 18 DNA in oral squamous cell carcinoma and normal mucosa

The aim of this study was to ascertain the prevalence of HPV 16/18 DNA in oral squamous cell carcinoma (OSCC) vs. normal oral mucosa, and to correlate the virologic data with other factors obtained from the patients' records. One hundred and thirteen paraffin embedded tissue samples (73 OSCC and 40 normal oral mucosa) were studied using HPV type specific primer-mediated polymerase chain reaction (PCR). Seventy-four per cent (54/73) of OSCC and 55% (22/44) of normal oral mucosa were positive for HPV 16/18 DNA. Statistical analysis indicated there was a significant difference between HPV16/18 positive OSCC vs. normal oral mucosa (P=0.040), and that age (<60 years) and gender (male) were correlated with the presence of HPV16/18 in the tumour. No significant association was found between the presence of HPV and other risk factors, including tobacco use, alcohol use, tumour location, histologic grade or TNM staging. We found a significant association of HPV16/18 with oral squamous cell carcinoma. Also, HPV16/18 is a co-factor in oral carcinogenesis, particularly in male patients and patients under the sixth decade. In addition, we found that HPV infection is a common event in the normal oral mucosa.     

Insulin‐like growth factor II mRNA‐binding protein 3 expression promotes tumor formation and invasion and predicts poor prognosis in oral squamous cell carcinoma

J Oral Pathol Med (2011) 40 : 699–705 Background: Insulin‐like growth factor II mRNA‐binding protein 3 (IGF2BP3), an oncofetal RNA‐binding protein, has been implicated in the enhancement of proliferation and invasion in various cancers. This study aimed to investigate the clinical significance and functional role of IGF2BP3 expression in oral squamous cell carcinoma (OSCC). Methods: IGF2BP3 expression in 93 OSCC patients was investigated using immunohistochemical staining and correlated with clinical parameters and patients’ survival. The effect of IGF2BP3 on cell invasion ability was evaluated by RNA interference in OSCC cell line. Results: High expression of IGF2BP3 in OSCC was significantly correlated with large tumor size and lymph node metastasis. Kaplan‐Meier analysis revealed that oral cancer patients with high IGF2BP3 expression had a significantly lower 5‐year survival (P = 0.0017). Multivariate analysis of clinical samples demonstrated IGF2BP3 to be an independent prognosis factor (P = 0.003). Moreover, the IGF2BP3 shRNA significantly suppressed the invasion ability of OSCC in vitro, and the knockdown of endogenous IGF2BP3 expression also inhibited tumor formation in vivo. Conclusions: IGF2BP3 enhances cell invasion ability and tumorigenicity in human OSCC in vitro and in vivo. IGF2BP3 is an independent prognostic factor in patients with OSCC. Targeting of IGF2BP3 could potentially suppress the tumor growth and metastasis to improve the outcome of patients with OSCC.     

口腔鳞状细胞癌患者颈部淋巴结特征与预后的关系

目的： 探讨口腔鳞状细胞癌（oral squamous cell carcinoma,OSCC）患者颈部阴性淋巴结临床、病理形态特征,并研究其与颈淋巴结转移、复发及总生存期的关系。方法： 回顾分析2017年1月—2021年1月新疆医科大学第一附属医院收治的100例OSCC患者的临床病理资料,研究终点事件为颈淋巴结转移、复发和死亡,采用Kaplan-Meier法绘制生存曲线,Cox生存分析法探讨不同因素与患者预后的关系。采用SPSS 21.0软件包对数据进行统计学分析。结果： 100例患者平均随访时间为30个月,失访10例。90例患者中,出现颈淋巴结转移、复发11例（12.2%）、死亡28例（31.1%）,阴性淋巴结出现窦组织细胞增生（sinus histiocytosis, SH）27例（30%）,3年无颈淋巴结转移复发率为89.4%,3年总生存率为67.3%。90例患者的Cox回归生存分析结果显示,分化程度、N分期、临床分期对总生存期有显著影响（P<0.05）;吸烟、初次颈淋巴结转移（N≠0）、SH、行双侧颈淋巴清扫术对颈淋巴结转移、复发有显著影响（P<0.05）,且吸烟、初次颈淋巴结转移、SH、行双侧颈淋巴清扫术患者的颈淋巴结转移、复发概率较高。结论： SH、初次颈淋巴结转移是导致OSCC患者颈淋巴结转移、复发的独立危险因素,N分期是导致患者死亡的独立危险因素。     

Role of lipocalin 2 and its complex with matrix metalloproteinase-9 in oral cancer

Oral Diseases (2012) 18 , 734–740 Objectives: Recent evidence demonstrated that lipocalin (LCN)2 is induced in many types of human cancer, while the detection of its complex with matrix metalloproteinase (MMP)‐9 is correlated with the cancer disease status. We attempted to evaluate plasma expressions of LCN2, MMP‐9, and their complex (LCN2/MMP‐9) during the diagnostic work‐up of patients with oral squamous cell carcinoma (OSCC) and investigated their correlations with disease progression. Methods: In total, 195 patients with OSCC and 81 healthy controls were recruited. Expression levels of LCN2, MMP‐9, and LCN2/MMP‐9 were determined with immunoenzymatic assays. Results: Patients with OSCC exhibited significantly higher levels of LCN2, MMP‐9, and LCN2/MMP‐9 compared with healthy controls (LCN2: P < 0.001; MMP‐9: P < 0.001; LCN2/MMP‐9: P < 0.01). Plasma levels of LCN2, MMP‐9, and LCN2/MMP‐9 in patients with OSCC were significantly correlated with each other and were associated with more‐advanced clinical stages (P < 0.05) and/or a larger tumor size (P < 0.05), but were not associated with positive lymph‐node metastasis or distal metastasis. Conclusion: Our results suggest that plasma levels of LCN2 and the LCN2/MMP‐9 complex may be useful in non‐invasively monitoring OSCC progression, while supporting their potential role as biomarkers of oral cancer disease status.     

Human papilloma virus types in the oral and cervical mucosa of HIV‐positive South African women prior to antiretroviral therapy

Background: To evaluate the prevalence of human papilloma virus (HPV) infection and types in the oral and cervix mucosa of treatment‐naïve HIV‐1‐positive women with CD4 counts less than 300 cells per ml with no HPV‐associated oral lesions. Methods: Oral epithelium was harvested from the buccal mucosa and lateral borders of the tongue and cervical samples were collected from the endocervical area of 30 women, 22–64 years old. Cytobrush Plus cell collectors were used for sampling both anatomical areas. Genital pathology, obstetric and gynaecological history, co‐morbid disease, hormone therapy, sexual behavior and smoking history were assessed via physical examination and clinical interviews. Special investigations included cervical Papanicolau smears, CD4 counts and HIV‐1 viral loads. The linear array HPV test was used to determine HPV genotypes present in the specimens. Results: Oral HPV were identified in 20% (n = 6) of the patients, of which two had infection with two HPV types. Genital HPV was found in 96.7% (n = 29) of the women, of which only 14 had cytological abnormalities on Papanicolau smear. Infection with multiple HPV types were present in 93.1% (n = 27) of the patients, with an average of four HPV types per individual. Conclusions: South African HIV‐positive women with CD4 counts less than 300 cells per ml have a significant risk of cervical HPV strains and multiple strain infection of the cervix. The prevalence of HPV in normal oral mucosa was low but high‐risk types were present. Limited correlation between oral HPV types and those identified in the cervical mucosa was found.     

p53 gene mutations and HPV infection in primary head and neck squamous cell carcinomas do not correlate with overall survival: a long term follow-up study

We analyzed specimens of head and neck squamous cell carcinomas (HNSCC) from 110 patients for p53 gene mutations, and 92 of them for human papillomavirus (HPV) infection, in order to evaluate the prognostic significance of these factors by comparison with clinical follow-up data. Mutations within the exons 5 to 8 of the p53 gene were found in 48 tumors (44%). Sequencing revealed in most cases mis-sense mutations (16/21). Frequency of p53 gene mutations was not related to the tumor stage or the presence of lymph node metastases. Of the 46 tumors that were analyzed by immunohistochemistry. 26 stained positively (56%). The number of positively stained nuclei increased slightly with decreasing differentiation of the tumors, whereas no correlation was found between tumor stage and immunoreactivity. An infection with the high-risk HPV types 16 and 18 could be detected in 39/92 tumor specimens (42%.). Follow-up data were obtained from 99 patients within a range of 2 to 112 months. No dependence of overall survival on the presence of p53 gene mutations or HPV infection could be observed. The absence of statistically significant correlations between p53 gene mutation and progressive disease, however, does not deny its putative relevance in early phases of tumor development.     

Oral squamous sell carcinoma in North-Eastern Hungary I. Epidemiology

The purpose of this study was to characterize the epidemiological data of oral squamous cell carcinoma (OSCC) patients in North-Eastern Hungary. The medical records of 119 randomly selected patients with OSCC admitted to the Department of Maxillofacial Surgery of the Faculty of Dentistry, University of Debrecen were reviewed. Cases were identified according to the epidemiological data, as to site, tumor size, lymph node involvement, clinical stage, histological differentiation, treatment obtained, recurrence of disease and survival rate. There was an obvious male predominance (male:female = 5.2:1). The median age at diagnosis was 57.4 years. The most common sites of OSCC were the floor of the mouth (27.7%), the lip (26.9%) and the tongue (22.7%). The majority of the patients (58,8%) presented with early-stage (I-II) disease at the time of diagnosis, and 48.6% had moderately differentiated tumors. The most frequent therapeutic modality used was surgery with or with out of radiotherapy. Roughly one fifth of the cases (20.7%) experienced a recurrence of disease during the follow-up period. The overall 5-year survival rate was 38.7% (stage I: 68.3%, stage II: 31.0%, stage III: 22.6%, stage IV: 11.1%). There was a correlation between survival and tumor size, lymph node metastasis, clinical stage and surgical treatment (p < or = 0.05, respectively). We found no significant correlation between histological differentiation and survival. Clinical stage exerts the most strongly significant impact on survival. Therefore, early detection and proper surgical intervention enhance patients' survival most effectively.     

Outcomes of oral squamous cell carcinoma in Taiwan after surgical therapy: factors affecting survival.

PURPOSE: The study goal was to determine which clinical features correlated with 5-year survival in patients surgically treated for oral squamous cell carcinoma (OSCC) in Taiwan. Patients and Methods: The records of 378 OSCC patients surgically treated with or without chemotherapy and radiotherapy were reviewed retrospectively. Their 5-year survival in relation to age, gender, tumor site, lymph node involvement, presence of distant metastasis, staging, differentiation, and risk factors, including betel quid (BQ) chewing, cigarette smoking, and alcohol consumption, was analyzed. RESULTS: The majority of the patients were men (male-to-female ratio, 5.87:1) with the mean age of 57.1 +/-11.7 years. Tumors occurred mainly at the buccal mucosa (BM) (100 of 378, 26.5%), gingiva (105 of 378, 27.8%), and tongue (103 of 378, 27.2%). Neck nodal metastasis occurred frequently at the floor of the mouth (in >60% of cases), followed by the gingiva (45.7%), buccal mucosa (34%), and tongue (20.4%), whereas early distant metastasis was rare (5.3%). There were 104 (27.5%) stage 1, 96 (25.4%) stage 2, 98 (25.9%) stage 3, and 80 (21.2%) stage 4 patients. OSCC at the BM and gingiva was most (and at the tongue least) associated with risk factors of BQ use and smoking. The 5-year survival was 75%, 65.6%, 49%, and 30% for patients with stage I, II, III, and IV, respectively. The size, nodal involvement, distant metastasis, staging, differentiation, and BQ use significantly affected the survival (P <.05, Kaplan-Meier analysis). BQ use also correlated most significantly with the younger age of occurrence of OSCC patients. CONCLUSIONS: Our data suggest that early treatment is the key to increasing the survival of OSCC patients. Periodic screening of high-risk populations for OSCC represents an urgent need in Taiwan.     

Impact of interleukin-8 gene polymorphisms and environmental factors on oral cancer susceptibility in Taiwan

Oral Diseases (2012) 18 , 307–314 Objectives: Interleukin‐8 (IL‐8), which is an angiogenic chemokine with a high expression level in tumor tissues, plays important roles in developing many human malignancies including oral squamous cell carcinoma (OSCC). This study was designed to examine the association of IL‐8 gene polymorphisms with the susceptibility and clinicopathological characteristics of OSCC. Methods: A total of 270 patients with OSCC and 350 healthy control subjects were recruited. Four single nucleotide polymorphisms (SNPs) of IL‐8 genes were analyzed using polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) genotyping analysis. Results: Results showed that four IL‐8 SNPs (?251 T/A, +781 C/T, +1633 C/T, and +2767 A/T) were not associated with oral cancer susceptibility as well as clinicopathological parameters. But among 345 smokers, IL‐8 polymorphisms carriers with betel quid chewing were found to have a 17.41‐ to 23.14‐fold risk to have oral cancer compared to IL‐8 wild‐type carriers without betel quid chewing. Among 262 betel quid chewers, IL‐8 polymorphisms carriers with smoking have a 10.54‐ to 20.44‐fold risk to have oral cancer compared to those who carried wild type without smoking. Conclusions: Our results suggest that the combination of IL‐8 gene polymorphisms and environmental carcinogens might be highly related to the risk of oral cancer.     

Prognostic importance of DNA from human papillomavirus in patients with oral squamous cell carcinoma

Background Survival of patients with oral squamous cell carcinoma (OSCC) is generally low, with the likelihood of locoregional recurrence or disease progression (LR/DP). Knowledge of prognostic factors for survival is key to achieving an understanding and increased survival. The present study aimed to identify prognostic factors for patients with OSCC, especially the presence of DNA from human papillomavirus (HPV).Material and Methods Retrospective cohort study including 119 patients with OSCC treated at the National Cancer Institute in Mexico City (2009-2013). Clinical information was obtained from patient records including LR/DP. Formalin-fixed, paraffin-embedded tissues were obtained and used for detecting DNA from different types of HPV. Potential prognostic factors for Overall Survival (OS) were analyzed using the Cox proportional hazards model.Results After model adjustment, factors associated with longer OS were a pre-treatment platelet count above 400,000/mm3 (HR=0.09, p=0.026) and response to primary treatment (HR=0.26, p=0.001). HPV DNA was present in 23 (19.3%) of the patients and importantly, type 16 found in 19 of them. Although survival of HPV-positive patients was longer, difference was not significant. However, among patients with LR/DP, HPV positivity was significantly associated with increased survival (HR=0.23, p=0.034). Importantly, survival was significantly different for HPV-positive patients with LR/DP > 6 months (HR=0.20, p=0.002), had higher absolute lymphocyte count at start of treatment (HR=0.50, p=0.028) or had local rescue treatment (HR=0.24, p=0.019).Conclusions Although HPV positivity was not associated with a longer OS of OSCC patients, a better prognosis was significantly associated with HPV positivity and recurring or progressing disease, particularly with HPV type 16. Key words:HPV, human papillomavirus, HPV-16, oral squamous cell carcinoma, oral cancer, survival, locoregional recurrence, disease progression.