A novel algorithm for reliable detection of human papillomavirus in paraffin embedded head and neck cancer specimen

Human papillomavirus type 16 (HPV16) plays a role in the development of a subgroup of head and neck squamous cell carcinomas (HNSCC). However, uncertainty exists about the true impact of HPV in this tumor type as conflicting reports have been published with prevalence rates from 0 to 100%. We aimed to find a detection algorithm of a biologically and thus clinically meaningful infection, applicable for high-throughput screening of frozen and formalin-fixed paraffin embedded (FFPE) specimens. By considering detection of HPV E6 oncogene expression in frozen biopsies as gold standard for a meaningful HPV infection, the value of several assays was evaluated on FFPE tumor specimens and sera of 48 HNSCC patients. The following assays were evaluated on FFPE tissue samples: HPV DNA general primer (GP)5+/6+ PCR, viral load analysis, HPV16 DNA FISH detection, HPV16 E6 mRNA RT-PCR, p16 immunostaining, and on corresponding serum samples detection of antibodies against the HPV16 proteins L1, E6 and E7. Comparing single assays on FFPE tissue samples detection of E6 expression by RT-PCR was superior, but application remains at present limited to HPV16 detection. Most suitable algorithm with 100% sensitivity and specificity appeared p16 immunostaining followed by GP5+/6+ PCR on the p16-positive cases. We show that clinically meaningful viral HPV infections can be more reliably measured in FFPE HNSCC samples in a standard and high throughput manner, paving the way for prognostic and experimental vaccination studies, regarding not only HNSCC, but possibly also cancer types with HPV involvement in subgroups such as penile and anal cancer.     

Combined analysis of HPV-DNA, p16 and EGFR expression to predict prognosis in oropharyngeal cancer

Molecular prognostic indicators for oropharyngeal squamous cell carcinoma (OSCC), including HPV-DNA detection, epidermal growth factor receptor (EGFR) and p16 expression, have been suggested in the literature, but none of these are currently used in clinical practice. To compare these predictors, 106 newly diagnosed OSCC for the presence of HPV-DNA and expression of p16 and EGFR were analyzed. The 5-year disease-free survival (DFS) and overall survival (OS) were calculated in relation to these markers and a multivariate Cox analysis was performed. Twenty-eight percent of the cases contained oncogenic HPV-DNA and 30% were positive for p16. The p16 expression was highly correlated with the presence of HPV-DNA (p < 0.001). Univariate analysis of the 5-year DFS revealed a significantly better outcome for patients with p16-positive tumors (84% vs. 49%, p = 0.009). EGFR-negative tumors showed a tendency toward a better prognosis in DFS (74% vs. 47%, p = 0.084) and OS (70% vs. 45%, p = 0.100). Remarkable and highly significant was the combination of p16 and EGFR expression status, leading to 5-year DFS of 93% for p16+/EGFR- tumors vs. 39% for p16-/EGFR+ tumors (p = 0.003) and to a 5-year OS of 79% vs. 38%, respectively (p = 0.010). In multivariate analysis p16 remained a highly significant prognostic marker for DFS (p = 0.030) showing a 7.5-fold increased risk for relapse in patients with p16-negative tumors. Our data indicate that p16 expression is the most reliable prognostic marker for OSCC and further might be a surrogate marker for HPV-positive OSCC. HPV+/p16+ tumors tended to have decreased EGFR expression, but using both immunohistological markers has significant prognostic implications.     

广东省潮汕地区单中心头颈部鳞状细胞癌患者人乳头瘤病毒初步检测分析

目的探讨广东省潮汕地区单中心头颈部鳞状细胞癌(HNSCC)患者人乳头瘤病毒(HPV)感染及亚型状况。方法收集2014年12月至2016年12月汕头大学医学院附属肿瘤医院167例HNSCC患者的肿瘤原发灶标本。采用免疫组织化学(IHC)法检测肿瘤组织中p16蛋白的表达,以肿瘤细胞p16蛋白阳性率≥76%为判断HNSCC存在HPV的依据,分析p16蛋白与患者临床病理因素的关系。采用原位杂交法(ISH)检测肿瘤组织中是否存在HPV 16、18 DNA;应用RNAscope法检测肿瘤组织中18种常见的高危HPV亚型(HPV HR 18)RNA的表达情况,分析p16蛋白阳性细胞比例≥50%的肿瘤组织中HPV HR 18阳性情况。结果患者p16蛋白强表达率为7.2%(12/167);低龄组(<50岁)p16蛋白强表达率高于高龄组(≥50岁)[17.2%(5/29)比5.1%(7/138),χ^2=5.321,P=0.021];口咽癌组p16蛋白强表达率高于非口咽癌组[29.4%(5/17)比4.7%(7/150),χ^2=14.019,P<0.01];性别、烟酒嗜好及肿瘤分期分层患者间p16蛋白强表达率差异均无统计学意义(均P>0.05)。ISH检测示,全部HNSCC原发灶均未发现HPV 16、18 DNA,重复实验结果一致。RNAscope法检测示,肿瘤细胞p16蛋白阳性率≥50%的19例患者中,3例(15.8%)肿瘤组织HPV HR 18 RNA阳性。结论潮汕地区HNSCC患者的HPV阳性率较低,以口咽癌患者最高,且呈年轻化趋势。潮汕地区HNSCC主要致HPV亚型可能并非HPV 16、18,而可能是包括HPV HR 18中的其他致病亚型。     

头颈部癌表皮生长因子受体表达及对预后判断价值

[目的]探讨表皮生长因子受体(EGFR)在头颈部癌组织中表达情况及其对预后判断的价值。[方法]从2002年12月至2003年12月对91例经病理证实的初治头颈部癌(HNC),其中鼻咽癌(NPC)87例,头颈部鳞癌(HNSCC)4例的活检标本,应用免疫组织化学技术检测EGFR在癌组织中的表达情况。对有完善病历资料的61例HNC(其中NPC58例),探讨EGFR表达与临床病理特点及生存关系。[结果]在91例HNC中EGFR阳性74例,总表达率81.3%。87例NPC中阳性71例,表达率81.6%;4例HNSCC中阳性3例,表达率75%。61例HNC中,EGFR表达与性别、年龄、总分期、T分期、N分期无相关性。EGFR阳性组与阴性组3年总生存率(OS)、无病生存率(DFS)、无区域复发生存率(LRFS)、无远处转移生存率(DMFS)分别为73.5%、63.3%、71.4%、65.3%和90.0%、90.0%、90.0%、90.0%,但单因素分析其差异无显著性。在其中58例NPC中,EGFR表达与性别、年龄、总分期、T分期、N分期无相关性。EGFR阳性组和阴性组3年OS、DFS、LRFS、DMFS分别为72.3%、63.6%、72.2%、63.8%和100.0%、100.0%、100.0%、100.0%。在单因素分析中,两组3年DFS和DMFS有显著性差异(P<0.05),但在多因素分析未发现有显著性差异。[结论]EGFR在HNC中表达率较高,过度表达者呈现不良预后的趋势,但未获得统计学差异。     

Human papillomavirus status in young patients with head and neck squamous cell carcinoma

The role of human papillomavirus (HPV) in head and neck squamous cell carcinoma (HNSCC) development has been recognized only in the last decade. Although younger patients develop HNSCC associated with HPV, the incidence in young patients has not been studied. Forty-five young HNSCC patients (<40 years) were tested for HPV and the expression of p16(ink4a) and p53 in tumor biopsies. The presence of HPV was correlated with the absence and presence of alcohol and tobacco exposure. Paraffin-embedded, archival biopsy materials from HNSCC of 45 patients younger than 40 years were analyzed. HPV subtypes were identified by PCR followed by genotyping. Expression of p16(ink4a) and p53 were determined by immunohistochemistry. Fourteen (31%) of the HNSCC specimens from 45 patients unequivocally exhibited HPV16 positivity. Sixty percentage of the oropharyngeal tumors and 5% of the oral cavity tumors were HPV16 positive. P16(ink4a) overexpression was detected in 93% of the HPV16-positive tumors. None of the HPV16 tumors showed p53 overexpression. There was no association of HPV positivity with (lack of) exposure to alcohol and smoking. HPV association was not exclusively detected in nonsmoking, nondrinking young HNSCC patients. The presence of p16(ink4a) accumulation and the absence of p53 overexpression are good surrogate markers for HPV-associated HNSCC.     

The prognostic significance of tumor human papillomavirus status for patients with anal squamous cell carcinoma treated with combined chemoradiotherapy

The prognostic relevance of tumor human papillomavirus (HPV) status in anal squamous cell carcinoma (SCC) had not been previously investigated, although its relevance to cervical, head and neck SCC is known. We retrospectively evaluated outcomes in 47 patients with anal SCC treated with combined chemoradiotherapy (CCRT) and determined tumor HPV status by HPV DNA chip method and p16 expression by immunohistochemistry (IHC) from paraffin-embedded tumor tissues. The median age was 65 years (range, 44-90 years). Sixteen (34%) patients were diagnosed with T stage 3 to 4, and 18 (38%) patients had regional nodal disease (N-positive). Thirty-five (75%) patients were HPV positive, and 31 (66%) patients were genotype 16 (HPV16-positive). Thirty-nine (83.0%) patients were positive for p16. After median follow-up of 51.7 months (range, 5.1-136.0 months), HPV16-positive group had significantly better 4-year progression-free survival (PFS, 63.1% vs. 15.6%, p < 0.001) and overall survival (84.6% vs. 39.8%, p = 0.008) than HPV genotype 16 negative (HPV16-negative) group. Patients with p16-positive tumor also had a better 4-year PFS (52.5% vs. 25.0%, p = 0.014) than those with p16-negative tumor. In multivariate analysis for PFS, N-positive and HPV16-negative were independent prognostic factors for shorter PFS. Comparing patterns of failure, time to loco-regional failure was statistically superior in HPV16-positive over HPV16-negative groups (p = 0.006), but time to systemic failure was not different (p = 0.098). Tumor HPV genotype 16 status is a prognostic and predictive factor in anal SCC treated with CCRT, and p16 expression determined by IHC might be advocated as a surrogate biomarker of HPV integration in anal SCC. Further studies are warranted.     

Concurrent radiotherapy plus epidermal growth factor receptor inhibitors in patients with human papillomavirus-related head and neck cancer

Purpose

Concurrent radio-chemotherapy (RT-CT) is the standard treatment for locally advanced head and neck squamous cell carcinoma (LA-HNSCC), but RT plus epidermal growth factor receptor (EGFR) inhibitors is an effective option when CT is not appropriate. Human papillomavirus (HPV) is associated with an improved prognosis in LA-HNSCC; however, it has not been fully studied as a prognostic factor after RT + EGFR inhibitors.

Experimental design

Immunohistochemical expression of p16INK4A and PCR of HPV16 DNA were retrospectively analyzed in tumor blocks from 52 stage III/IV LA-HNSCC patients treated with RT + EGFR inhibitors. Disease-free survival (DFS) and overall survival (OS) were analyzed by the Kaplan–Meier method.

Results

DNA of HPV16 was found in six of 52 tumors (12 %) and p16 positivity in eight tumors (15 %). After a median follow-up time of 45 months (6–110), p16-positive patients treated with RT + EGFR inhibitors showed an improved DFS (2-year DFS 75 vs. 44 %, HR 0.25, 95 % CI 0.06–0.99, p = 0.047) compared with p16-negative patients. These differences were outperformed when compared by HPV16 status (2-year OS rates of 83 vs. 58 %, HR 0.17, 95 % CI 0.02–0.99, p = 0.049 and 2-year DFS rates of 83 vs. 45 %, HR 0.17, 95 % CI 0.02–0.99, p = 0.049). In the Cox regression analysis with OS as the end point, ECOG 0–1 was the only prognostic factor independently associated with a good prognosis in the multivariable analysis.

Conclusion

In this study, p16/HPV16-positive patients with LA-HNSCC treated with RT + EGFR inhibitors showed a better survival, not confirmed in multivariate analysis.     

Survival of squamous cell carcinoma of the head and neck in relation to human papillomavirus infection: review and meta-analysis

Human papillomavirus (HPV) has been associated with head and neck squamous cell carcinomas (HNSCC), especially of the oropharynx, with highest distribution in the tonsils. HPV infection has been associated with improved outcome, although not all the studies show consistent results. The reason for this is not clear. We reviewed all published articles and conducted a meta-analysis on the overall relationship between HPV infection and overall survival (OS) and disease-free survival (DFS) in HNSCC. Patients with HPV-positive HNSCC had a lower risk of dying (meta HR: 0.85, 95% CI: 0.7-1.0), and a lower risk of recurrence (meta HR: 0.62, 95%CI: 0.5-0.8) than HPV-negative HNSCC patients. Site-specific analyses show that patients with HPV-positive oropharyngeal tumours had a 28% reduced risk of death (meta HR: 0.72, 95%CI: 0.5-1.0) in comparison to patients with HPV-negative oropharyngeal tumours. Similar observations were made for DFS (meta HR: 0.51, 95% CI: 0.4-0.7). There was no difference in OS between HPV-positive and negative non-oropharyngeal patients. The observed improved OS and DFS for HPV-positive HNSCC patients is specific to the oropharynx; these tumours may have a distinct etiology from those tumours in non-oropharyngeal sites.     

Patterns of antibody responses to nonviral cancer antigens in head and neck squamous cell carcinoma patients differ by human papillomavirus status

There have been hints that nonviral cancer antigens are differentially expressed in human papillomavirus (HPV)-positive and HPV-negative head and neck squamous cell carcinoma (HNSCC). Antibody responses (AR) to cancer antigens may be used to indirectly determine cancer antigen expression in the tumor using a noninvasive and tissue-saving liquid biopsy. Here, we set out to characterize AR to a panel of nonviral cancer antigens in HPV-positive and HPV-negative HNSCC patients. A fluorescent microbead multiplex serology to 29 cancer antigens (16 cancer-testis antigens, 5 cancer-retina antigens and 8 oncogenes) and 29 HPV-antigens was performed in 382 HNSCC patients from five independent cohorts (153 HPV-positive and 209 HPV-negative). AR to any of the cancer antigens were found in 272/382 patients (72%). The ten most frequent AR were CT47, cTAGE5a, c-myc, LAGE-1, MAGE-A1, -A3, -A4, NY-ESO-1, SpanX-a1 and p53. AR to MAGE-A3, MAGE-A9 and p53 were found at significantly different prevalences by HPV status. An analysis of AR mean fluorescent intensity values uncovered remarkably different AR clusters by HPV status. To identify optimal antigen selections covering a maximum of patients with ≤ 10 AR, multiobjective optimization revealed distinct antigen selections by HPV status. We identified that AR to nonviral antigens differ by HPV status indicating differential antigen expression. Multiplex serology may be used to characterize antigen expression using serum or plasma as a tissue-sparing liquid biopsy. Cancer antigen panels should address the distinct antigen repertoire of HPV-positive and HPV-negative HNSCC.     

HPV-Associated p16 INK4A Expression and Response to Therapyand Survival in Selected Head and Neck Cancers

Background: Development of squamous cell cancer of head and neck (SCCHN) is associated with humanpapillomavirus (HPV) infection, which in turn is closely related with expression of p16 INK4A. Loss of p16 INK4Aexpression by deletion, mutation, or hypermethylation is common in SCCHN. We here evaluated p16 INK4A as aprognostic marker of treatment response and survival in our SCCHN patients with laryngeal, hypopharyngeal ornasopharyngeal cancers. Materials and Methods: 131 patients diagnosed with SCCHN between January 2,2006and July 17, 2010 were examined for p16 INK4A. The median age was 60 years (15-82 years). Fifty one patients werestage I-II and 80 were stage III-IV. Immunohistochemical expression of p16 INK4A was analyzed in pretreatmentparaffin-embedded tumor blocks. The influence of p16 INK4A status on disease-free survival, and overall survivalafter treatment was evaluated. Results: P16 INK4A positivity was found in 58 patients (44%). Tumor-positivity forp16 INK4A was correlated with improved disease free survival (70.1 months vs 59 months) and improved overallsurvival (2, 3 and 5-year values; 77% vs 72%, 70% vs 63% and, 63% vs 55%; respectively). On multivariateanalysis, stage was determined as independent prognostic factor for disease-free survival. Conclusions: Stagewas the major prognostic factor on treatment response and survival in our patients. P16 INK4A status predictsbetter outcome in laryngeal, hypopharyngeal or nasopharyngeal cancer cases treated with surgery plus adjuvantradiochemotherapy as well as with definitive radiation therapy and/or chemotherapy.     

Impact of viral E2-gene status on outcome after radiotherapy for patients with human papillomavirus 16-positive cancer of the uterine cervix

PURPOSE: Integration of high-risk papillomavirus DNA has been considered an important step in oncogenic progression to cervical carcinoma. Disruption of the human papillomavirus (HPV) genome within the E2 gene is frequently a consequence. This study investigated the influence of episomal viral DNA on outcome in patients with advanced cervical cancer treated with primary radiotherapy. METHODS AND MATERIALS: Paraffin-embedded biopsies of 82 women with locally advanced cervical cancer could be analyzed for HPV infection by multiplex polymerase chain reaction (PCR) by use of SPF1/2 primers. E2-gene intactness of HPV-16-positive samples was analyzed in 3 separate amplification reactions by use of the E2A, E2B, E2C primers. Statistical analyses (Kaplan-Meier method; log-rank test) were performed for overall survival (OS), disease-free survival (DFS), local progression-free survival (LPFS), and distant metastases-free survival (DMFS). RESULTS: Sixty-one (75%) of 82 carcinomas were HPV positive, 44 of them for HPV-16 (72%). Seventeen of the 44 HPV-16-positive tumors (39%) had an intact E2 gene. Patients with a HPV-16-positive tumor and an intact E2 gene showed a trend for a better DFS (58% vs. 38%, p = 0.06) compared with those with a disrupted E2 gene. A nonsignificant difference occurred regarding OS (87% vs. 66%, p = 0.16) and DMFS (57% vs. 48%, p = 0.15). CONCLUSION: E2-gene status may be a promising new target, but more studies are required to elucidate the effect of the viral E2 gene on outcome after radiotherapy in HPV-positive tumors.     

Inverse relationship between human papillomavirus-16 infection and disruptive p53 gene mutations in squamous cell carcinoma of the head and neck.

PURPOSE: Squamous cell carcinomas of the head and neck (HNSCC) often harbor p53 mutations, but p53 protein degradation by the viral oncoprotein E6 may supercede p53 mutations in human papillomavirus 16 (HPV16)-positive tumors. The prevalence of p53 mutations in HPV-positive HNSCCs is indeed lower, but in some tumors these alterations coexist. The purpose of this study was to discern whether HNSCCs differ in the type of p53 mutations as a function of HPV16 status. EXPERIMENTAL DESIGN: The study was nested within a prospective multicenter study (ECOGE 4393/RTOG R9614) of patients with HNSCC treated surgically with curative intent. Tumors from one study center were used to construct a tissue microarray. The tumors were well characterized with respect to p53 mutational status. The tissue microarray was evaluated by HPV16 in situ hybridization. HPV16 analysis was also done on a select group of tonsillar carcinomas known to harbor disruptive p53 mutations defined as stop mutations or nonconservative mutations within the DNA binding domain. RESULTS: HPV16 was detected in 12 of 89 (13%) HNSCCs. By tumor site, HPV16 was detected in 12 of 21 (57%) tumors from the palatine/lingual tonsils, but in none of 68 tumors from nontonsillar sites (P < 0.00001). Both HPV16-positive and HPV16-negative HNSCCs harbored p53 mutations (25% versus 52%), but disruptive mutations were only encountered in HPV16-negative carcinomas. Of seven tonsillar carcinomas with disruptive p53 mutations, none were HPV16 positive, in contrast to HPV16-positive tonsillar carcinomas without disruptive p53 mutations (0% versus 57%; P = 0.008). CONCLUSIONS: Although HPV16 and mutated p53 may coexist in a subset of HNSCCs, HPV16 and disruptive p53 mutations seem to be nonoverlapping events. A less calamitous genetic profile, including the absence of disruptive p53 mutations, may underlie the emerging clinical profile of HPV16-positive HNSCC such as improved patient outcome.     

Influence of Tumor Site on Survival in Young Patients with Head and Neck Squamous Cell Carcinoma

The number of patients under the age of 45 diagnosed with head and neck squamous cell carcinomas (HNSCC) is increasing, probably due to the incidence of oropharyngeal cancers. Comparisons of HNSCC in young and old patients regarding tumor site and survival in sample sizes of relevance are rarely published. The aim of the study was to analyze the differences in survival between age groups dependent on tumor site and the influence of oropharyngeal cancers on the rising rates of HNSCC in the young. The records of 4466 patients diagnosed with HNSCC were reviewed retrospectively. Patients younger than 45 years were divided further into four subgroups for specific age differences in the young. The influences of patient and clinicopathological characteristics on survival were assessed using Kaplan–Meier analyses. Among the patient cohort, 4.8% were younger than 45 years. Overall survival (OS) in these patients was better, with a 5-year OS of 66.1% (vs. 46.4%), while relapse-free survival (RFS) was better in the older patient population, with a 5-year RFS of 74.9% (vs. 68.1%). Decreased RFS in the young was found for advanced tumor stages and tumor sited at the larynx. Hypopharynx and advanced stages were independent risk factors for OS under 45 years. Overall, 44.4% of all HNSCC in patients under 30 years were nasopharyngeal cancers, and incidence decreased with age. The incidence of oropharyngeal cancers increased significantly with age. Better OS in the young may be explained by lower tumor and disease stages, whereas oropharyngeal tumors and HPV were not found to cause rising rates of HNSCC. Laryngeal malignancies in young patients might be related to an increased malignant potential and should, consequently, be treated as such.     

Prognostic value of pretreatment and recovery duration of cranial nerve palsy in nasopharyngeal carcinoma

ABSTRACT: BACKGROUND: The purpose of this study was to evaluate the prognostic value of cranial nerve (CN) palsy in nasopharyngeal carcinoma (NPC) patients. METHODS: A retrospective analysis was performed on CN involvement using medical records of 178 consecutive patients with histologically diagnosed, non-disseminated NPC. RESULTS: In 178 NPC patients with CN palsy, the 5-year survival rates were as follows: overall survival (OS), 61.0%; disease-specific survival (DSS), 69.6%; local relapse-free survival (LRFS), 75.2%; distant metastasis-free survival (DMFS), 73.4%; and disease-free survival (DFS), 55.3%. Significant differences were observed in the 5-year OS rates between patients with single and multiple CN palsy (69.8% vs. 54.3%; P = 0.033) and the OS rates between patients with different pretreatment durations (68.7% vs. 43.3%, P = 0.007). However, no significant differences were observed in OS, DSS, LRFS and DFS rates between patients with upper and lower CN palsy (P = 0.581, P = 0.792, P = 0.729 and P = 0.212, respectively). The results showed that recovery duration was an independent prognostic factor for OS (HR = 2.485; P < 0.001), DSS (HR = 2.065; P = 0.016), LRFS (HR = 3.051; P = 0.001) and DFS (HR = 2.440; P < 0.001). CONCLUSIONS: Recovery duration is an independent prognostic factor for NPC patients with CN palsy and is related to recurrence, which leads to poor survival. Recovery duration requires close surveillance and different treatment regimens.     

31例头颈部骨外浆细胞瘤患者临床特点及预后分析

目的探讨头颈部骨外浆细胞瘤(EMP)的临床特点、治疗情况及预后相关因素。方法回顾性分析复旦大学附属眼耳鼻喉科医院2005年1月至2020年1月收治的31例头颈部EMP患者的临床资料。按病灶累及部位, 分为未累及韦氏环组和累及韦氏环组。采用单因素卡方检验分析组间差异, 采用Kaplan-Meier法分析总生存(OS)期。组间OS率和无局部复发生存(LRFS)率比较用log-rank检验。结果 31例患者全部接受了放射治疗, 中位随访时间98(7～192)个月。其中16例患者同时接受了手术, 10例患者同时接受了化疗, 2例患者同时接受了手术和化疗。15例患者累及韦氏环, 16例患者未累及韦氏环。10年OS率84.8%, 2例发生了局部复发, 2例发展为多发性骨髓瘤。年龄≤55岁和>55岁者10年OS率为100%∶67.0%(P=0.039);累及韦氏环者和未累及者10年OS率为93.3%∶75.2%(P=0.031), 二者10年LRFS率为100%∶66.7%(P=0.022)。放疗前肿瘤最长径≤5 cm者10年总生存率达95.0%, >5 cm者为35.7%(P=0.02...     

腋窝淋巴结1～3个转移的早期乳腺癌根治术后放疗的临床价值

Background and Objective:The role of postmastectomy radiotherapy (PMRT) in breast cancer patients with T1-T2 tumors and 1-3 positive axillary nodes is still uncertain. This study investigated the value of PMRT for these patients. Methods:In the retrospective data of 488 eligible patients, survival analysis was performed using the KaplanMeier method. Univariate and multivariate analyses were performed using a log-rank test and the Cox proportional hazards model, respectively. Results:The median observation time was 54 months. The 5- and 10-year Iocoregional recurrencefree survival (LRFS) rates were 90.8% and 86.9%, respectively. The 5- and 10-year disease-free survival (DFS) rates were 82.0% and 74.3%, respectively. The 5- and 10-year overall survival (OS) rates were 90.7% and 82.7%, respectively. For the 412 patients without PMRT, T2 classification, 2-3 positive nodes, and hormone (estrogen and progeeterone) receptornegative were risk factors for locoregional recurrence in the multivariate analysis. On the basis of these 3 risk factors, the group with 2-3 factors had a 10-year LRFS rate of 63.1% compared with 96.1% for the group with 0-1 factors (P<0.001＝.For the group with 2-3 risk factors, LRFS and DFS were significantly improved by PMRT, with the 5- and 10-year LRFS rates without PMRT of 82.4% and 63.1%, respectively, and, with PMRT, of 98.1% at both 5 years and 10 years (P =0.002). The 5- and 10-year DFS rates without PMRT were 72.0% and 57.6%, respectively, and, with PMRT, the 5- and 10-year DFS rates were 89.4% and 81.7%, respectively (P = 0.007). There was no significant difference in the 10-year OS rates between patients with and without PMRT. However, there is the potential benefit of 15.3% (87.1% vs. 71.8%, P =0.072). Conversely, the group with 0-1 factors of PMRT had no effect on prognosis. Conclusions:In patients receiving mastectomy with T1-T2 breast cancer with 1-3 positive nodes, for the group with 2-3 risk factors, PMRT significantly improved LRFS and DFS and has potential benefit in OS.     

Combined P16 and human papillomavirus testing predicts head and neck cancer survival

While its prognostic significance remains unclear, p16^INK4a protein expression is increasingly being used as a surrogate marker for oncogenic human papillomavirus (HPV) infection in head and neck squamous cell carcinomas (HNSCC). To evaluate the prognostic utility of p16 expression in HNSCC, we prospectively collected 163 primary tumor specimens from histologically confirmed HNSCC patients who were followed for up to 9.4 years. Formalin fixed tumor specimens were tested for p16 protein expression by immunohistochemistry (IHC). HPV type‐16 DNA and RNA was detected by MY09/11‐PCR and E6/E7 RT‐PCR on matched frozen tissue, respectively. P16 protein expression was detected more often in oropharyngeal tumors (53%) as compared with laryngeal (24%), hypopharyngeal (8%) or oral cavity tumors (4%; p < 0.0001). With respect to prognosis, p16‐positive oropharyngeal tumors exhibited significantly better overall survival than p16‐negative tumors (log‐rank test p = 0.04), whereas no survival benefit was observed for nonoropharyngeal tumors. However, when both p16 and HPV DNA test results were considered, concordantly positive nonoropharyngeal tumors had significantly better disease‐specific survival than concordantly negative nonoropharyngeal tumors after controlling for sex, nodal stage, tumor size, tumor subsite, primary tumor site number, smoking and drinking [adjusted hazard ratio (HR) = 0.04, 0.01–0.54]. Compared with concordantly negative nonoropharyngeal HNSCC, p16(+)/HPV16(?) nonoropharyngeal HNSCC (n = 13, 7%) demonstrated no significant improvement in disease‐specific survival when HPV16 was detected by RNA (adjusted HR = 0.83, 0.22–3.17). Our findings show that p16 IHC alone has potential as a prognostic test for oropharyngeal cancer survival, but combined p16/HPV testing is necessary to identify HPV‐associated nonoropharyngeal HNSCC with better prognosis.     

上皮-肌上皮癌的临床特点与疗效分析

目的 分析上皮-肌上皮癌患者局部区域复发风险和长期生存的相关因素。方法 回顾分析1999—2015年收治的18例首程治疗或首程辅助治疗于本中心完成的上皮-肌上皮癌患者生存情况。单纯手术8例(44%),术后辅助放疗9例(50%),根治性同步放化疗1例(6%)。比较各组局部区域复发率和生存率。Kaplan-Meier法计算生存率,logrank法检验。结果 中位随访时间46个月,全组5例(27.8%)患者LRR,5年LRFS率为69%,5年OS率为93%。辅助放疗组5年LRFS优于单纯手术组,但未达统计学差异(71%∶57%,P=0.569)。结论 局部区域失败是上皮-肌上皮癌治疗的主要失败模式,进一步提高LC率是改进生存的关键。     

同步放化疗联合热疗与同步放化疗治疗局部晚期宫颈癌的有效性和安全性比较

目的:比较后程密集热疗联合放化疗对比单独放化疗在局部晚期宫颈癌治疗中的安全性和有效性。方法:本研究收集了自2012年3月至2019年7月经我院治疗的局部晚期宫颈癌患者62例。其中32例患者接受了标准放化疗联合后程密集盆腔深部热疗,剩余30例患者接受了标准放化疗。两组放化疗方案一致:盆腔外照射（50 Gy/25 f）＋腔内放疗（A点剂量30 Gy／5 f）+同期紫杉醇联合铂类（TP）方案全身化疗。主要研究终点是完全缓解率和5年局部无复发生存率。次要研究终点是客观缓解率、5年无病生存率、5年总生存率和安全性。结果:统计学检查显示两组患者的基线特征是平衡的。研究结果显示热放化疗组的客观缓解率、5年无病生存率和5年总生存率均优于单独放化疗组（93.8% vs 90.0%,P=0.588;62.1% vs 47.4%,P=0.212;71.9% vs 58.8%,P=0.316）,尽管没有达到统计学差异。然而,热放化疗组的完全缓解率显著优于标准放化疗组（78.1% vs 53.3%,P=0.039）。热放化疗组的5年局部无进展生存率也显著高于标准放化疗组（72.9% vs 51.2%,P=0.048）。至于毒副反应,患者能够很好地耐受热放化疗。而且,相比单独放化疗,热放化疗有减轻中性粒细胞缺乏,恶心和乏力等毒副作用的趋势,且未增加额外的急性或者晚期毒副反应。结论:后程密集热疗联合同步放化疗对比单独同步放化疗显著提高了局部晚期宫颈癌患者的完全缓解率和5年局部无复发生存率。     

Systemic and local human papillomavirus 16-specific T-cell immunity in patients with head and neck cancer

Squamous cell carcinomas of the head and neck (HNSCC), in particular those of the oropharynx, can be caused by human papilloma virus Type 16 (HPV16). Whereas these HPV-induced oropharyngeal carcinomas may express the HPV16 E6 and E7 oncoproteins and are associated with better survival, the nonvirally induced HNSCC are associated with overexpression of p53. In this study we assessed the presence of systemic and local T cells reactive against these oncoproteins in HNSCC. An exploratory study on the presence, type and function of HPV16- and/or p53-specific T cells in the blood, tumor and/or metastatic lymph node as measured by several immune assays was performed in an unselected group of 50 patients with HNSCC. Tumor tissue was tested for HPV DNA and the overexpression of p53 protein. Almost all HPV16+ tumors were located in the oropharynx. Circulating HPV16- and p53-specific T cells were found in 17/47 and 7/45 tested patients. T cells were isolated from tumor cultures and/or lymph nodes of 20 patients. HPV16-specific T cells were detected in six of eight HPV+ tumors, but in none of the 12 HPV-tumors. Tumor-infiltrating p53-specific T cells were not detected. In depth analysis of the HPV16-specific T-cell response revealed that this response comprised a broad repertoire of CD4+ T-helper Type 1 and 2 cells, CD4+ regulatory T cells and CD8+ T cells reactive to HPV16. The local presence of HPV16-specific T-cell immunity in HPV16-induced HNSCC implicates a role in the antitumor response and support the development of immunotherapy for HNSCC.