Cyclin D(1) and D(3) expression in vestibular schwannomas

OBJECTIVES: The G1 regulators of the cell cycle, cyclin D(1) and D(3), have been implicated in the regulation of Schwann cell proliferation and differentiation. The purpose of this study is to evaluate cyclin D(1) and D(3) protein expression and the corresponding clinical characteristics of vestibular schwannomas. STUDY DESIGN AND METHODS: Tissue sections of 15 sporadic vestibular schwannomas were prepared. Immunohistochemical analysis of the vestibular schwannomas was performed with anticyclin D(1) and anticyclin D(3) antibodies. The immunoreactivity was evaluated in comparison with adjacent vestibular nerves. Tissue sections of breast carcinoma and prostate carcinoma were used as positive controls for cyclin D(1) and D(3) staining, respectively. Patient demographics, tumor characteristics, and cyclin D expression were reviewed, and statistical analysis was performed. RESULTS: While the breast carcinoma control expressed abundant cyclin D(1) protein, none of the 15 vestibular schwannomas showed detectable cyclin D(1) staining. In contrast, seven of 15 vestibular schwannomas stained positive for the cyclin D(3) protein. Cyclin D(3) staining was taken up in the nucleus of schwannoma tumor cells in greater proportion than Schwann cells of adjacent vestibular nerve. Although sample size was small, no significant difference in the average age of presentation, tumor size, and male to female ratios for the cyclin D(3)(+) or cyclin D(3)(-) groups was found. CONCLUSION: The Cyclin D(1) protein does not appear to play a prominent role in promoting cell cycle progression in vestibular schwannomas. In contrast, cyclin D(3) expression was seen in nearly half of the tumors examined, suggesting that it may have a growth-promoting role in some schwannomas. Further studies are needed to define its cellular mechanism.     

Cyclin D1 amplification and p16(MTS1/CDK4I) deletion correlate with poor prognosis in head and neck tumors

OBJECTIVES/HYPOTHESIS: Cyclin D1, a cell cycle regulator localized to chromosome 11q13, is amplified in several human tumors including head and neck squamous cell carcinoma (HNSCC). Amplification and/or overexpression of cyclin D1 have been correlated to a poor prognosis. Deletion of the p16 gene, localized to 9p21, has also been observed in a significant proportion of HNSCC. The p16 gene regulates cyclin D1-CDK4 activity and prevents retinoblastoma tumor suppressor gene phosphorylation, thereby downregulating cellular proliferation. Detection of cyclin D1 amplification and p16 deletion using a simple and sensitive method will be valuable for the development of effective treatment modalities for head and neck cancer. STUDY DESIGN: We have used fluorescence in situ hybridization (FISH) to study cyclin D1 amplification and p16 gene deletion in head and neck tumors. Both single- and dual-color FISH were performed. METHODS: Paraffin-embedded tissues from 103 patients with HNSCC were analyzed using genomic DNA probes for cyclin D1 and p16. Dual-color FISH was performed with chromosome 11 or 9 centromeric probes as a control. Twenty-eight of these samples were analyzed for p16 expression by immunohistochemistry. RESULTS: Cyclin D1 amplification was observed in 30% (31/103) of patients, and p16 deletion in 52% (54/103). Lack of p16 expression was observed in 64% (18/28) of patients. There was a good correlation between the deletion of p16 sequences and the loss of p16 expression (P = .008). Amplification of cyclin D1 had a statistically significant association with recurrence, distant metastasis, and survival at 36 months. There was a significant association between p16 deletion and the development of distant metastases. Cyclin D1 amplification and p16 deletion together correlated with recurrence, distant metastasis, and survival. CONCLUSIONS: We demonstrate that FISH is a simple and sensitive method for detecting cyclin D1 amplification and p16 deletion in head and neck cancer. Our results suggest that these two genetic aberrations together portend a poorer outcome than either of the abnormalities alone in head and neck cancer.     

细胞周期蛋白循环素在下咽癌生物学行为中的意义

目的:探讨细胞周期蛋白,循环素(CyclinD1)在下咽癌生物学行为中的意义。方法:应用免疫组织化学SP法检测CyclinD1在53例下咽鳞状细胞癌手术后标本,11例癌旁正常黏膜标本中的表达。结果:CyclinD1在下咽癌细胞中的表达高于正常黏膜(P<0.05),且为弥漫性细胞核表达,CyclinD1表达与T分级明显正相关,r=0.345;CyclinD1表达阳性率与原发部位有关,r=-0.386,梨状窝区阳性率最高;CyclinD1表达与下咽癌其他临床指标间经统计学处理差异无显著性。结论:CyclinD1表达与T分级及原发部位明显相关,T分级越高,阳性表达率越高,梨状窝区阳性率最高。     

细胞周期蛋白D1基因表达与喉癌生物学行为的关系

为探讨ｃｙｃｌｉｎ Ｄ１基因扩增的临床意义，采用免疫荧光技术检测了２５例喉鳞癌的新鲜标本。发现细胞周期蛋白Ｄ１基因在喉癌中有９例扩增，而且与肿瘤分级和ＴＮＭ分期差别无显著性意义；而与肿瘤组织相邻近的正常组织表达阴性，或仅见极个别散在表达，推测ｃｙｃｌｉｎ Ｄ１基因在喉癌的发生发展中可能是非早期现象。     

Beta-catenin and cyclin D1 in mucoepidermoid carcinoma of variable histologic grades

OBJECTIVE: To analyze the expression of beta-catenin and cyclin D1 in mucoepidermoid carcinoma (MEC) of variable histologic grades to establish a correlation between the expression of these proteins and the different histopathologic grades of this neoplasia. DESIGN: Immunohistochemical analysis of MEC. SETTING: Pathological Anatomy Service, Discipline of Oral Pathology, Department of Dentistry, Federal University of Rio Grande do Norte, Natal, Brazil. PATIENTS: Fifteen cases of MEC, graded and categorized according to criteria reported in the literature into 5 tumors with a low grade of malignancy, 4 with an intermediate grade, and 6 with a high grade. MAIN OUTCOME MEASURES: Labeling patterns of beta-catenin and cyclin D1. RESULTS: No significant difference in beta-catenin labeling patterns was observed between low- and intermediate-grade tumors or between low- and high-grade tumors (P = .60 and P = .06, respectively; Fisher exact test), despite a strong tendency toward a difference in the latter. In contrast, a significant difference was noted between intermediate- and high-grade tumors (P = .03). For cyclin D1, no labeling was observed in any high-grade cases, and only 3 cases showed overexpression of this protein. Comparison of the labeling patterns among the different histologic grades revealed no significant difference. CONCLUSIONS: The reduced expression of beta-catenin observed in all high-grade MECs is probably due to the loss of its adhesion function, which confers a greater invasive potential to these tumors. The overexpression of cyclin D1 observed in only 3 MEC cases suggests that this protein does not participate in the etiopathogenesis of these tumors, which implies that other genes are likely responsible.     

An immunohistochemical study of cyclin D1 protein expression in laryngeal squamous cell carcinoma

CONCLUSION: Contrary to most reports, our study shows that the expression of cyclin D1 is not an independent prognostic factor in patients with laryngeal cancer (LC). No correlation between cyclin D1 expression and survival rates in LC was found in a multivariate analysis. OBJECTIVES: The aim of this study was to determine the possible relevance of the expression of cyclin D1 protein in LC as prognostic criteria and to analyse correlation of the expression with clinicopathological features and survival rates. MATERIALS AND METHODS: Immunohistochemistry staining was used to detect the expression of cyclin D1 in 130 samples of laryngeal cancer and in 22 specimens of laryngeal nodules. RESULTS: Cyclin D1 expression was found in 52 (40%) LC samples and in 3 (13.6%) samples of laryngeal nodules. There was no significant correlation between cyclin D1 expression and clinicopathological features of LC. A multivariate analysis of survival confirmed that cyclin D1 expression was not an independent prognostic factor in LC.     

Proliferation potential in recurrent acoustic schwannoma following gamma knife radiosurgery versus microsurgery

OBJECTIVE: To evaluate the proliferation potential of recurrent acoustic schwannoma following gamma knife radiosurgery (GKR) versus microsurgery. STUDY DESIGN: Retrospective study. METHODS: A review of surgical records of the House Ear Clinic revealed 8 patients who had undergone GKR and 15 patients who had undergone microsurgery who had unilateral acoustic schwannoma recurrences. Immunohistochemical studies were performed to evaluate the expression of proliferating cell nuclear antigen (PCNA) on archival paraffin-embedded blocks. RESULTS: All 8 GKR and 15 microsurgical tumors had positive staining for PCNA. The recurrent GKR tumors had significantly lower proliferation levels than in the microsurgical group (P = .03). Two GKR tumors had high proliferation levels. CONCLUSIONS: Our study indicates that recurrent vestibular schwannomas treated with GKR have lower proliferation potential as assessed by PCNA compared with recurrences following microsurgery. Radiation-induced apoptosis is thought to contribute to the lower tumor cell proliferation in GKR tumor. The two GKR tumors with high proliferation potential could be a result of radiation-induced sporadic mutation, resulting in high tumor cell proliferation.     

A comparison of direct eighth nerve monitoring and auditory brainstem response in hearing preservation surgery for vestibular schwannoma.

OBJECTIVE: The objective of this study was to compare the effectiveness of direct eighth nerve monitoring (DENM) and auditory brainstem response (ABR) in facilitating hearing preservation during vestibular schwannoma resection. STUDY DESIGN: This was a retrospective study. SETTING:: Tertiary referral center. METHODS: We conducted a retrospective clinical study of the use of ABR and DENM during vestibular schwannoma removal. Tumors were removed through a retrosigmoid craniotomy. The rate of hearing preservation between the two monitoring modalities was compared. The additional outcome measures of facial nerve function and cerebral spinal fluid leak rate were also evaluated. RESULTS: Hearing preservation was attempted in 77 patients with vestibular schwannomas. Tumor sizes ranged from 0.5 cm to 2.5 cm. Hearing was preserved in 71% of patients with tumors 1 cm or less and in 32% of patients with tumors between 1 and 2.5 cm when direct eighth nerve monitoring was used. Hearing preservation rates with ABR for tumors 1 cm or less were 41% and 10% in patients with tumors between 1 and 2.5 cm (p=0.03) Facial nerve preservations rates were 94% (House-Brackmann 1-2) for tumors less than 2 cm. CONCLUSIONS: DENM provides significantly higher rates of hearing preservation during vestibular schwannoma resection when compared with ABR.     

Inactivation of p53 and Amplification of Cyclin D1 Correlate With Clinical Outcome in Head and Neck Cancer

The authors have investigated whether genetic abnormalities in two genes, loss of heterozygosity (LOH) of p53 and amplification of the cyclin D1 gene, correlate with clinical outcome in 56 matched pairs of blood and tumor from patients with squamous cell carcinoma of the head and neck (SCCHN). Frequency of p53 LOH was 47.4%, of cyclin D1 amplification 33.9%, and of both abnormalities together 23.7%. p53 LOH was associated with T4 (P = 0.003) and stage IV (P = 0.015) tumors. Cyclin D1 amplification was associated with recurrences and/or metachronous tumors (P = 0.007). The total number of p53 and cyclin D1 abnormalities (scored as zero, one, and two) show a pattern that seems to be additive; the increase in the number of these abnormalities is associated with a proportional increase in the frequency of T4, stage IV, presence of recurrences and/or metachronous tumors, and possibly a proportional decrease in the disease-free interval in the sample. The association of the markers with recurrences and/or metachronous tumors persists if the tumor stage effect is mathematically removed. The combined analysis of the p53 and cyclin D1 abnormalities seems to be more informative than either of them individually and may have predictive value in SCCHN.     

Association between high initial tissue levels of cyclin d1 and recurrence of nasopharyngeal carcinoma

OBJECTIVES/HYPOTHESIS: Cyclin D1 expression and the rate of apoptosis have been reported to serve as important prognostic indicators in human cancers. The purpose of the present study was to determine the prognostic significance of both initial cyclin D1 expression and the apoptotic index in nasopharyngeal carcinoma. STUDY DESIGN: Cohort study. METHODS: Cyclin D1 protein levels and apoptosis in tumors and their corresponding adjacent, histologically normal tissues were determined at the time of initial diagnosis using immunohistochemical staining, Western blot analysis, and in situ end labeling, respectively, in 64 patients with T1-T4/N0-N2, poorly differentiated squamous cell carcinoma of the nasopharynx. All cases were treated by routine radiation therapy with a total median dose of 70 Gy and followed up for 10 years. RESULTS: High levels of cyclin D1 were found in 35 of 64 tumor specimens (54.7%); no cyclin D1--positive cells were determinable in normal epithelium of the nasopharynx. Rates of early local recurrence (within 5 y) were significantly higher (P <.01) for patients with high levels of cyclin D1 before radiation therapy (24 of 35 patients [68.6%]) as compared with patients with low or no expression (3 of 29 [10.3%]). Furthermore, patients bearing high levels of cyclin D1 had a poorer prognosis concerning 10-year survival than the others (P <.001), whereas overexpression of cyclin D1 did not correlate with the initial TMN classification (P >.05). According to the rate of spontaneous apoptosis in tumors below or above the median, patients were divided into two groups. There was no statistically significant difference for the overall survival between the two groups (P >.05). CONCLUSIONS: The present study demonstrates that cyclin D1 can be used as an indicator of recurrence and subsequent prognosis in nasopharyngeal carcinoma after radiation therapy. At the same time, the apoptotic state before radiation therapy is of no value in predicting the prognosis of patients with nasopharyngeal carcinoma.     

MRI evaluation of neurofibromatosis 2 patients: a standardized approach for accuracy in interpretation.

OBJECTIVE: To determine the level of agreement between local radiologists' and an experienced neuroradiologist's measurements of vestibular schwannomas. STUDY DESIGN: Prospective study with uniform magnetic resonance acquisition protocol parameters and reporting instructions across 30 magnetic resonance imaging facilities worldwide. SETTING: Multicenter natural history study of neurofibromatosis Type 2. SUBJECTS: One hundred fifteen magnetic resonance imaging examinations of 57 neurofibromatosis Type 2 patients older than 5 years of age. INTERVENTIONS: Thin-slice, postcontrast cranial magnetic resonance imaging. MAIN OUTCOME MEASURES: Spearman's rho interobserver association coefficient of vestibular schwannoma linear measurements. RESULTS: The local and experienced radiologist measurements and identification of tumors agreement was fair (kappa = 0.77). Discordant interpretations were adjudicated by another experienced neuroradiologist. CONCLUSION: The least interobserver variability was found in measurements of thin-slice postcontrast magnetic resonance imaging scans obtained at neurofibromatosis Type 2 centers in patients without previous operations and moderately sized tumors. If the schwannoma was difficult to assess, because of magnetic resonance imaging acquisition protocol, postoperative changes, or tumors smaller than 5 mm in greatest diameter, the neuroradiologist provided a more thorough assessment. The authors suggest uniform reporting criteria for vestibular schwannoma assessments to ensure clinically relevant information is communicated regarding vestibular schwannoma size.     

Cycin D1在鼻腔鼻窦肿瘤中的表达

目的：探讨cycin D1在鼻腔鼻窦肿瘤中的表达及其临床意义。方法：采用免疫组化SABC法检测20例鼻腔鼻窦恶性肿瘤和10例相应部位良性病变的石蜡标本中cycin D1的表达。结果：鼻腔鼻窦肿瘤中cycin D1表达均为阳性,良性病变中为阴性或弱阳性。鼻腔鼻窦恶性肿瘤中cycin D1积分与患者年龄、性别、病变部位及病理类型无关（P＞0.05）。Cycin D1积分与TNM分期、病理分级相关（P＜0.05 ),且积分随肿瘤临床分期、分级逐渐升高。初诊病例与复发病例之间cyclin D1积分差异有统计学意义（P＜0.05 ）。结论：Cycin D1在鼻腔鼻窦肿瘤的发生、发展及复发转移等临床病理过程中起重要作用。     

An immunohistochemical study of cyclin D1 protein expression in laryngeal squamous cell carcinoma

Conclusion. Contrary to most reports, our study shows that the expression of cyclin D1 is not an independent prognostic factor in patients with laryngeal cancer (LC). No correlation between cyclin D1 expression and survival rates in LC was found in a multivariate analysis. Objectives. The aim of this study was to determine the possible relevance of the expression of cyclin D1 protein in LC as prognostic criteria and to analyse correlation of the expression with clinicopathological features and survival rates. Materials and methods. Immunohistochemistry staining was used to detect the expression of cyclin D1 in 130 samples of laryngeal cancer and in 22 specimens of laryngeal nodules. Results. Cyclin D1 expression was found in 52 (40%) LC samples and in 3 (13.6%) samples of laryngeal nodules. There was no significant correlation between cyclin D1 expression and clinicopathological features of LC. A multivariate analysis of survival confirmed that cyclin D1 expression was not an independent prognostic factor in LC.     

Simultaneous contralateral vestibular schwannoma and glomus jugulare tumor: a case report

To the best of our knowledge, only 3 cases of a simultaneous vestibular schwannoma and a glomus jugulare tumor have been previously reported in the literature. In all 3 cases, the lesions were located on the same side. We report a new case of simultaneous vestibular schwannoma and glomus jugulare tumor that is unique in that the two lesions arose on opposite sides. The glomus tumor was treated with embolization followed by radiotherapy, while the schwannoma was managed via radiologic observation.     

PD-L1及CD8^+TIL在前庭神经鞘膜瘤中的表达及意义

目的探讨程序性死亡蛋白配体1(PD-L1)及CD8+肿瘤浸润性淋巴细胞(Tumor infiltrating lymphocyte,TIL)在散发性前庭神经鞘膜瘤(vestibular schwannoma,VS)中的表达,为前庭神经鞘膜瘤免疫靶向治疗提供理论和实验基础。方法回顾性收集2018年11月至2019年4月于上海交通大学医学院附属第九人民医院耳鼻咽喉头颈外科手术治疗的15例散发性VS患者的临床资料及病理蜡块,采用免疫组化法检测15例散发性前庭神经鞘膜瘤手术切除组织中PD-L1及CD8^+淋巴细胞表达情况,分析它们之间相关性及与临床病理特征的关系。结果显微镜下观察15例散发性VS组织中有14例(93.3%)PD-L1表达阳性,其中7例(46.7%)为高表达。在15例(100%)样本中均观察到CD8^+淋巴细胞的阳性表达,其表达量与PD-L1表达量呈正相关(P=0.02,r=0.82)。结论在散发性VS中存在PD-L1广泛表达,PD-L1表达与CD8^+TIL表达呈正相关,该肿瘤可能对免疫检查点抑制剂的免疫治疗有反应,在未来临床试验中可进一步探索。     

Neurotrophic factor expression in vestibular schwannoma. An overview

The vestibular schwannoma is a benign, slow-growing neoplasm that originates from the neurolemmal sheath of the vestibular branch of the VIIIth cranial nerve. This tumor entity accounts for 6 % of all intracranial tumors and the annual incidence of newly diagnosed vestibular schwannoma is reported as 13 per million. The molecular pathogenesis of both sporadic vestibular schwannoma and those occurring in neurofibromatosis type II appears to be associated with an aberration of a tumor suppressor gene on chromosome 22q12. The biological background for the various growth patterns of vestibular schwannoma is, however, largely unknown. This differing clinical and biological behaviour of vestibular schwannoma may be explained by the presence of neurotrophic factors. The results of recent immunohistochemical studies demonstrate the co-expression of transforming growth factor (TGF)-beta 1 and glial cell line-derived neurotrophic factor (GDNF) in vestibular schwannoma and suggest a trophic synergism of both neurotrophic factors in this tumor. Moreover, expression of numerous different neurotrophic factors has been shown in studies of nerve growth factor (NGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), fibroblast growth factor (FGF), neuregulin (NRG) and erythropoietin (EPO) indicating a biological role in development, maintainance or growth of vestibular schwannoma. In this article, we summarize the findings on neurotrophic factor expression and discuss their characteristics and biological role in vestibular schwannoma.     

Loss of alleles in vestibular schwannomas: use of microsatellite markers on chromosome 22

OBJECTIVE: Using highly informative microsatellite markers flanking the neurofibromatosis type 2 gene, we determined the frequency of chromosome 22 allele loss in vestibular schwannomas. DESIGN: Peripheral lymphocyte/vestibular schwannoma DNA pairs were analyzed with five different microsatellite markers on chromosome 22. PATIENTS: Samples were taken from 32 patients (17 females and 15 males). Twenty-seven tumors occurred sporadically, and five were from patients with neurofibromatosis type 2. RESULTS: Using the microsatellite markers D22S351, CRYB2, D22S268, D22S304, and interleukin type 2RP3, we found loss of heterozygosity for at least two markers in 12 tumors. Ten tumors showed loss of heterozygosity for markers flanking the neurofibromatosis type 2 gene. Although microsatellite markers require little DNA for analysis and are highly informative, allele patterns may be difficult to interpret in some cases. CONCLUSIONS: Loss of heterozygosity of chromosome 22 alleles was a frequent event in vestibular schwannomas. In 10 tumors, heterozygosity was lost for centromeric and telomeric markers indicating likely monosomy 22. However, 63% of tumors did not reveal a detectable chromosomal loss. Unless a second vestibular schwannoma locus exists, these tumors likely harbor point mutations in the neurofibromatosis type 2 gene or deletions below the level of resolution of the markers used in this study.     

Postoperative magnetic resonance imaging findings after transtemporal and translabyrinthine vestibular schwannoma resection

OBJECTIVES/HYPOTHESIS: Magnetic resonance imaging (MRI) has become the investigation of choice to follow up patients after vestibular schwannoma resection. STUDY DESIGN: Retrospective. METHODS: Postoperative MRI findings of 70 patients after vestibular schwannoma resection through a transtemporal (n = 48) and a translabyrinthine (n = 22) approach were reviewed. Time-dependent changes in intensity, size, and shape of enhancement in the internal auditory canal before and after contrast administration, postoperative temporal lobe gliosis, and changes of fat grafts were evaluated. RESULTS: After vestibular schwannoma resection, all patients showed signal enhancements in the internal auditory canal ranging from a faint to high signal intensity in the first postoperative MRI, 3 to 6 months after surgery. In the next MRI at 12 to 24 months after surgery, 30 patients (43%) showed a decreased signal, 35 patients (50%) a stable enhancement, and 5 patients (7%) an increased enhancement in the internal auditory canal depicted as an intense nodular or mass-like pattern. In patients with decreased or stable enhancement, a residual tumor could be excluded in the following MRI scans, whereas in all patients with increased enhancements after 12 to 24 months, signal enhancement further increased and residual tumors were detected. Different degrees of temporal lobe gliosis were found in 15 of 48 cases (31%) after transtemporal tumor removal. Enhancement of fat grafts used in 22 cases decreased to different degrees in 14 cases (64%). CONCLUSIONS: Differentiation of residual tumor from scar tissue in the internal auditory canal after vestibular schwannoma resection requires close, long-term follow-up. Nodular and progressive enhancements in the internal auditory canal indicate residual tumor. Linear enhancement in the internal auditory canal has been found to be a common finding after vestibular schwannoma resection not associated with residual tumor.     

喉鳞癌组织中CDK_4、cyclin D_1、P~(16)蛋白表达

目的：细胞周期中G1／S转换受到G1期的细胞周期蛋白及细胞周期蛋白的依赖激酶（CDKs）和它们的抑制剂的调节,cyclinD1／CDK4复合物磷酸化（pRB）基因产物、P16与cyclinD1竞争性地抑制CDK4,这些蛋白质的不正常表达就使细胞增殖失控,进而导致肿瘤的形成和发展。本研究从细胞周期调控的角度来探讨喉鳞状细胞癌的发生、发展与CDK4、cyclinD1及P16的关系。方法：用SP免疫组织化学的方法,检测了105例喉癌（其中声门上癌51例,声门癌54例）及28例声带息肉中CDK4、cyclinD1、P16蛋白表达的水平。结果显示：在喉癌组织中cyclinD1的表达以＋＋～＋＋＋为主,而在声带息肉组织中以阴性及弱阳性为主。它们之间存在着显著性的差异（P＜0．05）;而同样P16的表达在喉癌及对照组之间的差异非常显著（P＜0．001）。P16、CDK4的表达在声门上癌与声门癌之间差异显著,P值分别为P=0．002,P=0．021。经Spearman相关分析得出：喉癌组织中CDK4的表达与P16的表达和cyclinD1的表达具有相关性,相关系数分别为0．235、0．253。伴转移的喉癌组织中CDK4的表达与cyclinD1的表达相关明显,相关系数r=0．534。而在非转移的喉癌组织中CDK4与P16相关,相关系数r=0．253。由此我们推论：在喉癌的发生、发展中CDK4、cyclinD1及P16是共