Multimodality functional imaging using DW-MRI and 18F-FDG-PET/CT during radiation therapy for human papillomavirus negative head and neck squamous cell carcinoma: Meixoeiro Hospital of Vigo Experience

AIM To noninvasively investigate tumor cellularity measured using diffusion-weighted magnetic resonance imaging(DW-MRI) and glucose metabolism measured by 18 Flabeled fluorodeoxyglucose positron emission tomography/computed tomography(18F-FDG-PET/CT) during radiation therapy(RT) for human papillomavirus negative(HPV-) head and neck squamous cell carcinoma(HNSCC).METHODS In this prospective study, 6 HPV- HNSCC patients underwent a total of 34 multimodality imaging examinations(DW-MRI at 1.5 T Philips MRI scanner [(n = 24) pre-, during-(2-3 wk), and post-treatment(Tx), and 18F-FDG PET/CT pre- and post-Tx(n = 10)]. All patients received RT. Monoexponential modeling of the DW-MRI data yielded the imaging metric apparent diffusion coefficient(ADC) and the mean of standardized uptake value(SUV) was measured from 18F-FDG PET uptake. All patients had a clinical follow-up as the standard of care and survival status was documented at 1 year.RESULTS There was a strong negative correlation between the mean of pretreatment ADC(ρ =-0.67, P = 0.01) and the pretreatment 18F-FDG PET SUV. The percentage(%) change in delta(?) ADC for primary tumors and neck nodal metastases between pre- and Wk2-3 Tx were as follows: 75.4% and 61.6%, respectively, for the patient with no evidence of disease, 27.5% and 32.7%, respectively, for those patients who were alive with disease, and 26.9% and 7.31%, respectively, for those who were dead with disease.CONCLUSION These results are preliminary in nature and are indicative, and not definitive, trends rendered by the imaging metrics due to the small sample size of HPV- HNSCC patients in a Meixoeiro Hospital of Vigo Experience.

Role of human papillomavirus in oropharyngeal squamous cell carcinoma: A review

Human papillomavirus(HPV) has been implicated in the pathogenesis of a subset of oropharyngeal squamous cell carcinoma. As a result, traditional paradigms in relation to the management of head and neck squamous cell carcinoma have been changing. Research into HPVrelated oropharyngeal squamous cell carcinoma is rapidly expanding, however many molecular pathological and clinical aspects of the role of HPV remain uncertain and are the subject of ongoing investigation. A detailed search of the literature pertaining to HPV-related oropharyngeal squamous cell carcinoma was performed and information on the topic was gathered. In this article, we present an extensive review of the current literature on the role of HPV in oropharyngeal squamous cell carcinoma, particularly in relation to epidemiology, risk factors, carcinogenesis, biomarkers and clinicalimplications. HPV has been established as a causative agent in oropharyngeal squamous cell carcinoma and biologically active HPV can act as a prognosticator with better overall survival than HPV-negative tumours. A distinct group of younger patients with limited tobacco and alcohol exposure have emerged as characteristic of this HPV-related subset of squamous cell carcinoma of the head and neck. However, the exact molecular mechanisms of carcinogenesis are not completely understood and further studies are needed to assist development of optimal prevention and treatment modalities.

Clinical and scientific impact of human papillomavirus on head and neck cancer

Head and neck cancer(HNC) arises from the skull base to the clavicles and is the fifth most common cancer in the world by incidence. Historically, in the developed world HNC was associated with tobacco use and alcohol consumption, and the combination of the two produced a synergistic increase in risk. However, beginning in 1983, investigators have found a significant and growing proportion of HNC patients with human papillomavirus-positive(HPV) tumors who neither drank nor used tobacco. Since that time, there has been increased interest in the molecular biology of HPV-positive HNC. Multiple studies now show that HPV has shifted the epidemiological landscape and prognosis of head and neck squamous cell carcinoma(HNSCC). These studies provide strong evidence for improved survival outcomes in patients with HPV-positive HNSCC compared to those with HPV-negative HNSCC. In many reports, HPV status is the strongest predictor of locoregional control, disease specific survival and overall survival. In response to these findings, there has been significant interest in the best management of HPV-positive disease. Discussions within major cooperative groups consider new trials designed to maintain the current strong survival outcomes while reducing the long-term treatment-re-lated toxicities. This review will highlight the epidemiological, clinical and molecular discoveries surrounding HPV-related HNSCC over the recent decades and we conclude by suggesting how these findings may guide future treatment approaches.

Advanced oropharyngeal squamous cell carcinoma: Pathogenesis, treatment, and novel therapeutic approaches

Oropharyngeal cancer accounts for approximately 2.8% of newly cancer cases. Although classically a tobacco related disease, most cases today are related to infection with human papilloma virus(HPV) and present with locally advanced tumors. HPV related tumors have been recognized as a molecularly distinct entity with higher response rates to therapy, lower rates of relapse, and improved overall survival. Treatment of oropharyngeal cancer entails a multi-disciplinary approach with concomitant chemoradiation. The role of induction chemotherapy in locally advanced tumors continues to be controversial however large studies have demonstrated no difference in survival or time to treatment failure. Surgical approaches may be employed with low volume oropharyngeal cancers and with development new endoscopic tools, more tumors are able to be resected via an endoscopic approach. Given advances in the understanding of HPV related oropharyngeal cancer, ongoing research is looking at ways to minimize toxicities via de-intensification of therapy. Unfortunately, some patients develop recurrent or metastatic disease. Novel therapeutics are currently being investigated for this patient population including immunotherapeutics. This review discusses the current understanding of the pathogenesis of oropharyngeal cancer and treatment. We also discuss emerging areas of research as it pertains to de-intensification as well novel therapeutics for the management of metastatic disease.

Prognostic Significance of Vascular Endothelial Growth Factor-A (VEGF-A) and Ki-67 Expression in Head and Neck Cancer Patient with Negative Neck

Background: Management of N0 neck in patients with head and neck squamous cell carcinoma (HNSCC) remains a subject of continued debate. Prognostic biomarkers might provide useful information for treatment selection and adjustment. Objective: To evaluate the prognostic relevance of VEGF-A and Ki-67 expression to types of neck management. Methods: This prospective study included 140 patients with HNSCC. Tumor expression of VEGF-A and Ki-67 was measured by immunohistochemistry. Based on tumor size and site criteria, 88 patients with N0 neck were categorized as high, intermediate and low risk of subclinical neck diseases and accordingly treated by elective neck dissection (END), irradiation (ENI) and observation. Adjuvant treatment was given to tumor with close or positive margins. A multivariate Cox regression model was used to identify prognostic factors. Impact of biomarker expression, treatment type and risk category on disease-specific survival (DSS) in the setting of N0 neck were evaluated by Kaplan-Meier survival and adjusted hazard ratio (HR). Results: Coexpression of VEGF-A and Ki-67 (HR = 2.351, p = 0.021) and positive node (HR = 2.301, p = 0.009) were independent prognostic factors for HNSCC. In the setting of N0 neck, marker coexpression has an HR of 4.97 (p = 0.004) independent of treatment modalities (p = 0.069) and risk categories (p = 0.971). Alternatively, neither marker expression was predictive of a better treatment outcome for END compared to ENI, as suggested by the odds of patients being survived 15.4 times greater (p = 0.01) and the 5-year DSS rates of 85.1% versus 44.7% (p = 0.008). Conclusion: Coexpression of VEGF-A and Ki-67 is a suggestion of tumor microinvasiveness in addition to risk of lymph node metastasis and may indicate the need of adjuvant treatment despite negative tumor margins. Neither marker expression serves an indicator for the selection of END over ENI in neck management.

No evidence of HPV DNA in esophageal squamous cell carcinoma in a population of Southern Brazil

AIM:To investigate the association between human papillomavirus(HPV)and esophageal squamous cell carcinoma(ESCC)in southern Brazil.METHODS:We studied 189 esophageal samples from125 patients from three different groups:(1)102 biopsies from 51 patients with ESCC,with one sample from the tumor and another from normal esophageal mucosa distant from the tumor;(2)50 esophageal biopsies from 37 patients with a previous diagnosis of head and neck squamous cell carcinoma(HNSCC);and(3)37 biopsies from esophageal mucosa with normal appearance from 37 dyspeptic patients,not exposed to smoking or alcohol consumption.Nested-polymerase chain reaction(PCR)with the MY09/11 and GP5/6 L1primers was used to detect HPV L1 in samples fixed in formalin and stored in paraffin blocks.All PCR reactions were performed with a positive control(cervicovaginal samples),with a negative control(Human Genomic DNA)and with a blank reaction containing all reagents except DNA.We took extreme care to prevent DNA contamination in sample collection,processing,and testing.RESULTS:The histological biopsies confirmed the diagnosis of ESCC in 52 samples(51 from ESCC group and 1 from the HNSCC group)and classified as well differentiated(12/52,23.1%),moderately differentiated(27/52,51.9%)or poorly differentiated(7/52,13.5%).One hundred twenty-eight esophageal biopsies were considered normal(51 from the ESCC group,42 from the HNSCC group and 35 from dyspeptic patients).Nine had esophagitis(7 from the HNSCC and 2 from dyspeptic patients).Of a total of 189 samples,only 6 samples had insufficient material for PCR analysis:1 from mucosa distant from the tumor in a patient with ESCC,3from patients with HNSCC and 2 from patients without cancer.In 183 samples(96.8%)GAPDH,G3PDH and/orβ-globin were amplified,thus indicating the adequacy of the DNA in those samples.HPV DNA was negative in all the 183 samples tested:52 with ESCC,9 with esophagitis and 122 with normal esophageal mucosa.CONCLUSION:There was no evidence of HPV infection in different ESCC from southern Brazil.

Id2 regulates the proliferation of squamous cell carcinoma in vitro via the NF-κB/Cyclin D1 pathway

Squamous cell carcinoma (SCC) is a significant cause of cancer morbidity and mortality worldwide, with an incidence of up to 166 cases per 100 000 population. It arises in the skin, upper aerodigestive tract, lung, and cervix and affects more than 200 000 Americans each year. We report here that a microarray experiment comparing 41 SCC and 13 normal tissue specimens showed that Id2, a gene that controls the cell cycle, was significantly up-regulated in SCC. Enforced expression of Id2 in vitro stimulated the proliferation of SCC cells and up-regulated the transcription of nuclear factor kappa B (NF-κB) and cyclin D1. Enhancement of the NF-κB activity with p65 significantly increased the cell proliferation and the transcription of cyclin D1, whereas inhibition of the NF-κB activity with I kappa B alpha mutant (IκBα M) and pyrroline dithiocarbamate (PDTC) abrogated cell proliferation and transcription of cyclin D1. Furthermore, a mutated NF-κB binding site in the cyclin D1 promoter fully abrogated the Id2-induced transcription of cyclin D1. Taken together, these data indicate that Id2 induces SCC tumor growth and proliferation through the NF-κB/cyclin D1 pathway.

Programmed death 1 (PD-1) and ligand (PD-L1) inhibitors in head and neck squamous cell carcinoma: A meta-analysis

Background: PD-1 and PD-L1 inhibitors have emerged as promising treatments for patients with head and neck squamous cell carcinoma (HNSCC).Methods: Systematic review and meta-analysis of PD-1 and PD-L1 inhibitors in HNSCC. Outcomes: median overall survival (mOS), median progression-free survival (mPFS), Response Evaluation Criteria in Solid Tumors (RECIST) and treatment-related adverse events (TRAEs).Results: Eleven trials reported data on 1088 patients (mean age: 59.9 years, range: 18-90). The total mOS was 7.97 months (range: 6.0-16.5). Mean mPFS for all studies was 2.84 months (range: 1.9-6.5). PD-1 inhibitors had a lower rate of RECIST Progressive Disease than PD-L1 inhibitors (42.61%, 95% confidence interval [CI]: 36.29-49.06 vs. 56.79%, 95% CI: 49.18-64.19,P < 0.001). The rate of TRAEs of any grade (62.7%, 95% CI: 59.8-65.6) did not differ.Conclusions: Meta-analysis shows the efficacy of PD-1 and PD-L1 inhibitors in HNSCC and suggests a possible difference in certain RECIST criterion between PD-1 and PD-L1 inhibitors. Future work to investigate the clinical significance of these findings is warranted.

程序性死亡受体/配体免疫治疗策略在人乳头瘤病毒阳性头颈部鳞状细胞癌中的研究进展

人乳头瘤病毒(HPV)感染已成为头颈部鳞状细胞癌(HNSCC)的主要致病因素之一,与HPV阴性相比,HPV阳性HNSCC表现出以程序性死亡受体(PD)为代表的多种免疫细胞及其效应分子的表达增加。PD-1/PD-L1高表达的患者与HPV阳性HNSCC患者存活率的显著提高相关。而HPV阳性HNSCC患者在接受抗PD-1/程序性死亡配体(PD-L1)免疫治疗后的客观缓解率、无进展生存期、总生存期等指标较HPV阴性HNSCC患者有所提高,提示HPV阳性HNSCC患者在接受抗PD-1/PD-L1免疫治疗上可能取得更好疗效。另外,PD-1/PD-L1抑制剂联合HPV癌症疫苗、双通路抑制剂等免疫治疗方案在HPV相关癌症中已发挥出独特的优势。根据HPV状态为患者制定个体化免疫治疗是一种有前景的治疗策略。

基于影像组学鉴定头颈部鳞状细胞癌患者HPV状态:诊断性meta分析

目的:采用诊断性meta方法,评价影像组学特征,构建模型,鉴定头颈部鳞状细胞癌患者人乳头瘤病毒(HPV)状态的价值。方法:检索PubMed、Embase、Web of Science、中国知网、维普数据库中有关影像组学鉴定HPV状态的相关文献。检索时间为建库至2022年5月28日。采用Revman5.4等统计软件,评估预测模型的诊断效能。结果:共纳入9篇文献,包括3 312例患者。合并结果显示,基于影像组学方法诊断患者HPV状态的敏感度为0.82 [95%CI(0.74,0.87)],特异度为0.66 [95%CI (0.54,0.75)],阳性似然比为2.4 [95%CI (1.8,3.1)],阴性似然比为0.28 [95%CI(0.21,0.36)],诊断比值比为9[95%CI(6,12)],综合受试者工作特征曲线下面积(AUC)为0.82[95%CI(0.78,0.85)]。通过影像组学方法鉴定HPV状态先验概率为25%,可提升至44%(后验概率)。结论:采用影像组学方法鉴定头颈部鳞状细胞癌患者HPV状态有较高的实用价值。

宫颈鳞癌组织中miR-590-3P、FoxM1及Cdc25B细胞核内蛋白水平与高危型HPV感染的相关性研究

目的:研究宫颈鳞癌组织中微小RNA-590-3P(miR-590-3P)、叉头盆蛋白质M1(FoxM1)及细胞分裂周期因子25B(Cdc25B)细胞核内蛋白水平与高危型人乳头状瘤病毒(HPV)感染的相关性。方法:将2014年2月—2019年12月医院通过宫颈活检、锥切以及子宫切除获取的宫颈组织标本300份纳入研究。将其分为正常组织59例,宫颈上皮内瘤变(CIN)1组织62例,CIN2/3组织101例,宫颈鳞癌组织78例。采用实时荧光定量PCR检测不同组织中的miR-590-3P表达水平,以免疫组织化学法检测FoxM1、Cdc25B蛋白表达情况,并进行23种基因型HPV DNA的检测。比较不同宫颈组织中的miR-590-3P以及FoxM1、Cdc25B蛋白表达情况,高危型HPV感染情况,并以Spearman相关性分析明确宫颈鳞癌组织中的miR-590-3P以及FoxM1、Cdc25B蛋白表达情况与高危型HPV感染的关系。采用受试者工作特征(ROC)曲线明确miR-590-3P以及FoxM1、Cdc25B蛋白诊断CIN2+的效能。结果:CIN2/3组织以及宫颈鳞癌组织中的miR-590-3P表达水平均高于正常组织、CIN1组织,且CIN1组织中的miR-590-3P表达水平均高于正常组织(F=46.873,P=0.000);CIN2/3组织以及宫颈鳞癌组织中的FoxM1蛋白阳性率及Cdc25B蛋白阳性率均高于正常组织及CIN1组织,且CIN1组织中的FoxM1、Cdc25B蛋白阳性率均高于正常组织(χ^(2)=47.293、51.041,P=0.000、0.000)。CIN2/3组织以及宫颈鳞癌组织中的高危型HPV感染率明显高于正常组织、CIN1组织,且CIN1组织中的高危型HPV感染率明显高于正常组织(χ^(2)=67.293,P=0.000)。经Spearman相关性分析可得:宫颈鳞癌组织中miR-590-3、FoxM1、Cdc25B蛋白表达率均和高危型HPV感染呈正相关。经ROC曲线分析发现:miR-590-3P以及FoxM1、Cdc25B蛋白联合检测诊断CIN2+的曲线下面积、灵敏度、特异度分别为0.884、0.90、0.88,均高于上述三项指标单独检测。结论:宫颈鳞癌组织中miR-590-3P、FoxM1及Cdc25B细胞核内蛋白水平与高危型HPV感染密切相关,有望成为早期诊断CIN2+的潜在生物学指标,值得临床重点关注。

头颈部鳞癌人乳头状瘤病毒16/18感染状态与Ki-67、P53表达的关系

目的通过测定人头颈部鳞状细胞癌组织中人乳头状瘤病毒(HPV)16/18感染和Ki-67、P53蛋白的表达,探讨HPV16/18感染和Ki-67、P53蛋白表达的相关性及意义。方法采用荧光定量PCR方法测定头颈部鳞癌患者肿瘤组织中HPV16/18DNA的含量,同时以免疫组织化学方法测定Ki-67、P53蛋白的表达状态。结果 62.8%的患者在头颈部鳞癌组织中检测到HPV16/18DNA,46.15%的患者肿瘤组织表达P53蛋白为阳性。患者肿瘤组织Ki-67的表达在2%～70%。HPV16/18DNA含量与患者的性别、吸烟、饮酒和临床分期无相关性(P>0.05),与病理学分级呈正相关(r=0.350,P=0.001),与Ki-67表达无相关性(P=0.179),与P53表达呈负相关(r=-0.197,P=0.04)。结论头颈部鳞癌的发生可能和HPV16/18感染有关,致病机制涉及P53蛋白的降低表达。

E1A基因对头颈部淋巴结转移鳞癌细胞体外生长的抑制和化放疗增敏的作用

背景与目的:腺病毒5型早期区1A基因(early region 1A,E1A)是新近发现的一个具有抑癌作用的基因,E1A蛋白能够从正、负两种途径调控多种基因的表 达,具有诱导肿瘤细胞分化、逆转其恶性表型、降低其体内致瘤性和抗转移等活性,但E1A基因在头颈淋巴结转移鳞癌治疗中的作用尚少见报道。本研究的目的是探讨E1A基因对头颈淋巴结转移鳞癌细胞体外生长的抑制作用和化、放疗增敏作用及其初步的作用机理。方法:经脂质体介导,将真核细胞高效表达E1A基因的重组质粒peDNA3-E1A导入人舌鳞癌的淋巴结转移癌细胞系686LN-1。通过测绘生长曲线和计算倍增时间,观察E1A基因对该细胞系生长的抑制作用。用MTT法测定转染E1A基因前后,癌细胞对顺铂、泰素、5-氟尿嘧啶和博来霉素等化疗药物以及放疗敏感性的改变。流式细胞术和免疫细胞化学染色检测转染前后的细胞周期分布以及p53和HER-2/neu表达情况。结果:转染E1A基因细胞群体生长缓慢,倍增时间延长(是转染空载体细胞的1.48倍)。与686LN-1-vect比较,686LN-1-E1A细胞对顺铂、博来霉素、泰素和射线的敏感性(IC_(50))分别提高了8倍、20倍、10倍和1倍。细胞周期出现G_2/M期阻滞;免疫细胞化学染色显示E1A基因能抑制HER-2/neu的表达。

人食管鳞状细胞癌标本中乳头状瘤病毒DNA的检测

目的食管鳞状细胞癌有明显的地域性差异,它的发生与遗传、环境、饮食和某些微生物的感染有密切的关系,本研究通过检测食管鳞状细胞癌和相应正常食管组织中 HPV-11型和HPV-16型的 DNA,阐明 HPV 与食管鳞状细胞癌的关系.方法用 PCR 的方法检测22位食管鳞状细胞癌患者手术切除的癌组织和相应癌旁正常食管粘膜(共44例标本)中的HPV-11型和 HPV-16型 DNA 的存在情况,并对其中一例 HPV阳性肿瘤标本的 HPV 序列进行测定.结果在本组所涉及的22位患者共计44例标本中,对于HPV-11型,12例癌组织为阳性;有7例癌旁组织为阳性;癌组织和癌旁组织均为阳性的有6例.对于 HPV-16型,6例癌组织为阳性;6例癌旁组织为阳性;癌组织和癌旁组织均为阳性的有3例.序列分析结果表明所测定的序列与 GeneBank 登录的NC001525.1(HPV-11)、M14119.1(HPV-11)和 AF217526.1(HPV-11)的同源性极高,均为99%.结论通过实验结果分析,我们认为食管鳞状细胞癌与 HPV-11型和 HPV-16型主要衣壳蛋白 L1基因密切相关,HPV 可能在一定的地域环境中与食管鳞状细胞癌的发生具有相关性.

RNA干扰hTERT基因治疗喉鳞状细胞癌的实验研究

目的:探讨RNA干扰hTERT(人端粒酶逆转录酶)基因对人喉鳞状细胞癌的治疗作用。方法:根据hTERTcDNA序列构建表达hTERTmRNA特异的、含荧光素基因的shRNA真核表达质粒pshRNA1、pshRNA2。将shRNA质粒分别转染人喉癌Hep-2细胞株及荷瘤裸鼠瘤体内。以激光共聚焦显微镜观察质粒在Hep-2细胞及瘤体内的表达;以MTT法观察质粒对Hep-2细胞增殖的抑制作用;以蛋白印迹法(Westernblot法)检测Hep-2细胞中hTERT蛋白的表达;以免疫组化SP法检测裸鼠瘤体内hTERT蛋白的表达。结果:pshRNA1、pshRNA2转染Hep-2细胞及pshRNA1转染瘤体后,共聚焦显微镜下见大量的癌细胞表达绿色荧光,hTERT蛋白表达明显下降。细胞受pshRNA1、pshRNA2转染后,其生长活性受到明显抑制。体内抑瘤实验表明,与空质粒载体组(pshRNA4)和生理盐水组相比,pshRNA1组移植瘤生长明显受到抑制。结论:表达shRNA的、hTERT基因特异的真核表达质粒能有效转染体内、外喉鳞癌细胞,并可有效抑制人喉鳞癌细胞的生长,为今后应用RNA干扰基因治疗喉癌提供重要的实验参考。

头颈部鳞癌中HPV感染与HIF-1α、VEGF表达的关系及预后意义

目的分析人乳头瘤病毒(HPV)感染与缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)在头颈部鳞癌(HNSCC)组织中的表达,探讨HPV感染与HIF-1α、VEGF表达的关系及临床意义。方法对43例HNSCC标本采用原位杂交法检测HPV 6/11、16/18和31/33的感染情况;免疫组织化学法检测HIF-la和VEGF的表达。采用Spearman法进行等级相关分析,Kaplan-Meier法计算生存率,Cox模型多因素预后分析。结果 43例HNSCC患者中,HPV阳性感染者占25.6%,HIF-1α和VEGF的阳性表达率分别为65.1%和51.2%。HIF-1α的表达与T、N分期及临床分期有关(P<0.05),VEGF表达与N分期和复发有关(P<0.05)。相关分析显示,HNSCC组织中HIF-1α与VEGF表达呈正相关(r=0.359,P<0.05)。HPV阳性感染组与阴性感染组中HIF-1α和VEGF的表达率差异无统计学意义(P<0.05)。HPV阳性感染组和阴性感染组3年生存率分别为60.0%和59.7%(P>0.05),HIF-1α阳性表达组和阴性表达组3年生存率分别为49.9%和80.0%(P=0.060),VEGF阳性表达组和阴性表达组3年生存率分别为41.2%和81.0%(P<0.05)。结论 HIF-1α、VEGF可作为评估HNSCC发展、转移及预后的参考指标之一,尚不能明确HNSCC组织中HPV感染与HIF-1α、VEGF表达存在相关性。

人乳头瘤状病毒在头颈部鳞状细胞癌中的研究进展

高危型人乳头瘤状病毒是宫颈癌的主要致病因素,近年来研究发现人乳头瘤状病毒感染与头颈部鳞状细胞癌的发生也存在密切关系。本文对人乳头瘤状病毒相关性头颈部鳞状细胞癌的流行病学、致癌机制、诊断方法、治疗及预后等方面的研究进展作一综述。

两种检测头颈部鳞状细胞癌HPV16/18感染状态方法的对比

目的比较荧光定量PCR法(RT-PCR)和原位杂交法检测头颈部鳞状细胞癌中HPV(HPV)16/18亚型感染的差异。方法采用RT-PCR法和原位杂交法对78例头颈部鳞状细胞癌患者的肿瘤组织进行HPV感染状态的检测,评价两种方法的一致性。结果 RT-PCR法检测到62.8%的头颈部鳞癌组织中含有HPV16/18 DNA。原位杂交法检测到47.4%的肿瘤组织中有HPV16 DNA。用RT-PCR方法,唇、口腔、口咽及下咽的HPV16/18 DNA阳性率分别为33.3%、66.67%、70%和57.14%;用原位杂交方法,唇、口腔、口咽、下咽的HPV16 DNA阳性率分别为33.3%、43.8%、60.0%和57.1%。总体上,两种方法检测HPV16/18DNA具有较高一致性(Kappa=0.595,P<0.001)。结论 RT-PCR和原位杂交法检测HPV16/18 DNA结果的一致性较高。

对头颈鳞状细胞癌中高表达基质金属蛋白酶基因具有激活作用的人类膜蛋白基因筛选

目的对头颈部鳞状细胞癌中高表达的基质金属蛋白酶(MMP)基因(包括MMP1、MMP2、MMP3、MMP9、MMP10、MMP12和MMP13)具有激活作用的人类膜蛋白基因进行筛选。方法构建上述7个备选MMP启动子报告基因质粒。用豆蔻酸佛波酰乙酯(PMA,一种蛋白激酶C激活剂)刺激7个备选MMP启动子报告基因质粒。用双荧光素酶检测系统检测各备选MMP启动子报告基因质粒PMA刺激前的活性及PMA刺激后的活性,以后者除以前者得出各备选MMP启动子报告基因质粒的相对荧光素酶活性值(RLA),RLA最大者选定为MMP启动子报告基因质粒。用人类膜蛋白基因质粒库中的722种质粒依次刺激MMP启动子报告基因质粒,刺激前后MMP启动子报告基因质粒RLA均数超过10确定为对MMP启动子报告基因质粒活性有明显激活作用的人类膜蛋白基因质粒。将筛选出的人类膜蛋白基因质粒以相同方法进行第二轮筛选,第二轮筛选出的人类膜蛋白基因质粒以相同方法进行第三轮筛选,最终确定对头颈部鳞状细胞癌中高表达的MMP有激活作用的人类膜蛋白基因。结果PMA刺激后MMP1、MMP2、MMP3、MMP9、MMP10、MMP12、MMP13启动子报告基因质粒的RLA分别为4.53±0.72、1.47±0.28、23.28±1.96、2.37±0.46、8.81±1.35、2.15±0.51、3.47±0.99。MMP3启动子报告基因质粒的RLA明显高于其他MMP启动子报告基因质粒(P均<0.05),被选为MMP启动子报告基因质粒。全部722个质粒经3轮筛选,确定AXL受体酪氨酸激酶(AXL)、肿瘤坏死因子受体超家族成员10b(TNFRSF10B)和G蛋白耦联受体65(GPR65)质粒对MMP启动子报告基因质粒均有明显的激活作用。结论 GPR65、AXL、TNFRSF10B基因对头颈部鳞状细胞癌中高表达的MMP基因有激活作用。

头颈部鳞癌发病机制与治疗研究进展

头颈部鳞状细胞癌约占全身恶性肿瘤的6%,全球每年新发病例超过50万例。对其发病机制,近年来有一些新的发现,人乳头瘤病毒是除烟酒因素外另一个重要的头颈癌致病因素,表皮生长因子受体(EGFR)的信号传导与鳞癌的发生和转移亦有密切联系。Ⅰ、Ⅱ期头颈癌患者,通过单纯手术或放疗,均可取得较好效果。以多西他赛、顺铂、氟尿嘧啶为主的化疗药物,与调强放疗及不同分割方式联合的同期化放疗方案,是Ⅲ、Ⅳ期晚期口咽癌、下咽癌及喉癌的重要治疗手段。作用于EGFR靶点的西妥昔单抗,无论是单药或与放疗联合,均显示了很好的应用前景,在临床上逐步被广泛接受,提示靶向联合治疗可作为肿瘤手术后新的辅助治疗方法。Ⅲ、Ⅳ期局部晚期头颈癌患者的多中心临床试验尚在进行中,其联合治疗方案有待统一。