软骨肉瘤中p16蛋白表达的临床意义

目的：探讨p１６蛋白表达在软骨肉瘤诊断、分级和预后中的临床意义。方法：分析p１６蛋白表达与良、恶性软骨肿瘤和软骨肉瘤病理分级间以及与软骨肉瘤不同临床病理因素间的关系。结果：⑴p１６表达与软骨肉瘤组织学分级显著相关（Ｐ〈０．０５）,且在软骨肉瘤不同组织学分级间p１６蛋白表达差异有显著性（Ｐ〈０．０５）。⑵p１６表达在软骨肉瘤体积、复发方面差异有显著性（Ｐ〈０．０５）。结论：⑴p１６在软骨肉瘤形成和发展中     

Sox9 在人普通型与去分化型软骨肉瘤差异表达*

目的:探讨Sox9 在人普通软骨肉瘤,去分化软骨肉瘤和正常软骨中的表达。方法:取12例2003年1 月至2007年1 月在北京大学人民医院经病理切片确诊为软骨肉瘤（普通软骨肉瘤6 例,去分化软骨肉瘤6 例）及正常软骨组织（6 例）的新鲜冰冻标本,利用基因芯片的方法分析后,将Sox9 作为候选基因,而后利用Real-time PCR,Western blot和免疫组化的方法比较其在人普通软骨肉瘤,去分化软骨肉瘤和正常软骨组织中的差异表达。结果:基因芯片结果显示:与正常软骨组织相比,Sox9 在普通软骨肉瘤中上调约1.6 倍,在去分化软骨肉瘤中,其表达水平为正常软骨组织表达水平的0.082 倍;Real-time PCR检测结果显示:Sox9mRNA 在普通软骨肉瘤和去分化软骨肉瘤中表达水平分别为1.68± 0.119 和0.088 ± 0.017;Western blot分析表明Sox9 蛋白在人普通软骨肉瘤的表达量明显高于正常软骨组织,在去分化软骨肉瘤中的表达量显著低于正常软骨组织。免疫组化实验结果表明与正常软骨组织相比,6 例普通软骨肉瘤均有Sox9 表达,细胞染色呈较强的阳性,而去分化软骨肉瘤未发现较强阳性细胞出现,只有可疑的阳性细胞散在出现。结论:Sox9 在普通软骨肉瘤中的表达水平明显高于正常软骨组织,在去分化软骨肉瘤中的表达水平显著低于正常软骨组织。Sox9 在去分化软骨肉瘤中低水平表达与去分化软骨肉瘤疾病进展快,预后差呈正向相关。      

Runx2 在去分化软骨肉瘤细胞系中的表达及其意义

 目的　检测Runx2基因在去分化软骨肉瘤中与普通软骨肉瘤中的表达差异,进一步明确Runx2在去分化软骨肉瘤发生发展中的意义。方法　培养去分化软骨肉瘤细胞系NDCS-1及普通软骨肉瘤细胞系SW1353,提取细胞mRNA及总蛋白,RT-PCR、Western blotting及细胞免疫学检测Runx2在细胞系中的表达,之后对病理证实的去分化软骨肉瘤进行免疫组织化学染色,检测其在组织中的表达。结果　RT-PCR和Western blotting结果显示与普通软骨肉瘤细胞系SW1353相比,Runx2在去分化软骨肉瘤细胞系NDCS-1中高表达;免疫组织化学结果显示与普通软骨肉瘤成分相比,Runx2 在组织中的高度恶性成分中高表达。结论　Runx2在去分化软骨肉瘤中的高表达参与了去分化软骨肉瘤的发生和发展。     

p16表达和DNA倍体在软骨肉瘤中的临床意义

目的 探讨p16表达对软骨肉瘤诊断，分级和预后的临床意义。方法 应用免疫组化（S－P法）检测p16在软骨肉瘤石蜡切片中的表达，再用流式细胞仪检测软骨肉瘤DNA倍体。分析p16蛋白表达与良、恶性软骨肿瘤和软骨肉瘤病理分级间以及与软骨肉瘤不同临床病理因素的关系。结果 （1）p16表达与软骨肉瘤组织学分级显著相关（P＜0.05)。（4）p16表达在软骨肉瘤体积、复发差异有显著性（P＜0.05)。结论 p16蛋白表达对软骨肉瘤组织学分级，恶性进展和预后判断有指导意义。     

胸腔内去分化软骨肉瘤1例

去分化软骨肉瘤(dedifferentiated chondrosarcoma,DDCS)是一种较为罕见的原发性骨恶性肿瘤.去分化软骨肉瘤发病部位以股骨、胫骨最为多见,其次是骨盆,指(趾)骨少见,发生在肋骨者较少见报道.现报道杭州师范大学附属医院2010年7月收治的1例胸腔内肋骨起源的去分化软骨肉瘤的诊治经过,以提高临床医生对该病的认识.     

p16蛋白和p27蛋白在恶性骨肿瘤中的表达及其与预后的关系

目的 探讨p16和p27蛋白在恶性骨肿瘤组织中的表达及其与肿瘤的发生、发展及预后的关系.方法 应用免疫组织化学S-P法,检测48例软骨肉瘤、68例骨肉瘤,20例软骨瘤做对照;对p16蛋白在软骨肉瘤及p27蛋白在骨肉瘤不同病理分级、临床分期、预后分组中的表达进行对比分析和相关研究.结果 软骨肉瘤组织中p16蛋白高表达( )率与软骨瘤相比,差异有统计学意义(P＜0.01).软骨肉瘤复发组p16蛋白的高表达率与未复发组相比,差异有统计学意义(P＜0.05).p16蛋白高表达率与瘤组织分化程度呈正相关(P＜0.01),在软骨肉瘤不同组织学分级间p16蛋白高表达率相比差异均有统计学意义(P＜0.05).骨肉瘤组织中p27蛋白高表达( )率与软骨瘤相比,差异有统计学意义(P＜0.01).骨肉瘤低生存组p27蛋白高表达率与高生存组相比,差异有统计学意义(P<0.01).p27表达与瘤组织分化程度呈正相关(P＜0.01),与临床分期呈负相关(P＜0.01).结论 p16蛋白减少或缺失是软骨肉瘤发生、发展及不良预后的重要指标,p27蛋白减少或缺失是骨肉瘤的发生、发展及不良预后的重要指标,对临床具有重要的指导意义.     

LDN-193189对人去分化软骨肉瘤细胞系NDCS-1 的抑制作用研究*

目的:检测骨形成蛋白（BMP ）受体抑制剂LDN-193189对人去分化软骨肉瘤（DDCS）细胞系NDCS- 1 的抑制作用,探讨LDN-193189对去分化软骨肉瘤的抑癌机制。方法:以5 nmol/L 的LDN-193189作用于NDCS- 1 细胞,MTT 、平板克隆法检测LDN-193189对NDCS- 1 细胞的增殖抑制作用,Transwell 法、划痕实验检测LDN193189对NDCS- 1 细胞的侵袭抑制作用,Westernblot检测BMPR 2、p-Smad1/ 5 及RUNX 2 的蛋白表达抑制情况。结果:药物处理后NDCS- 1 细胞增殖、侵袭被明显抑制;药物处理后NDCS- 1 细胞的BMPR 2、p-Smad1/ 5 及RUNX 2 蛋白表达下降。结论:LDN-193189通过抑制BMPR 2-p-Smad 1/ 5-RUNX2 信号传导通路能有效抑制去分化软骨肉瘤细胞系NDCS- 1 的增殖侵袭能力。      

结直肠癌的微卫星不稳定与染色体不稳定

结直肠癌中存在微卫星不稳定（MSI）和染色体不稳定（CIN）。微卫星不稳定指微卫星序列中重复单位的获得或丢失，多由于DNA错配修复系统改变所致。染色体不稳定包括整条染色体的获得或缺失，染色体易位、重排等，结直肠癌中常见有1p和8p的删除、17p和18q的杂合性缺失以及20q的扩增。两种类型的结直肠癌具有不同的临床病理特征，在基因表达、疗效和预后等方面的差异性有待进一步研究。     

颌骨软骨肉瘤增殖细胞核抗原表达的免疫组化定量分析

增殖细胞核抗原（ｐｒｏｌｉｆｅｒａｔｉｎｇｃｅｌｌｎｕｃｌｅａｒａｎｔｉｇｅｎ,ＰＣＮＡ）是一种与细胞增殖周期有关的标志物,其表达状况与肿瘤细胞增殖活性相关。采用抗ＰＣＮＡ单克隆抗体（ｍＡｂ）定位和定量研究其表达,可作为判断细胞增殖情况及预后的一种手段。软骨肉瘤是颇为常见的原发性恶性骨肿瘤,其发病率仅次于骨肉瘤。本实验用免疫组化方法观察ＰＣＮＡ在软骨肉瘤的表达情况并定量分析,明确软骨肉瘤细胞的分化和增殖活性以及ＰＣＮＡ与其临床病理分级的关系。１　材料和方法１．１　标本来源２０例颌骨软骨肉瘤标…     

基因芯片筛选软骨肉瘤去分化相关基因初步探讨

目的：探讨用基因芯片技术区分普通型软骨肉瘤与去分化软骨肉瘤组织中基因的差异表达。方法：对病理证实的普通型软骨肉瘤及去分化软骨肉瘤组织,提取细胞mRNA,用寡核苷酸芯片,与正常关节软骨组织杂交后获得荧光信号,计算机软件分析荧光信号结果,并对数据进行归一化处理,分析组间显著变化的基因,然后对其进行聚类和主成份分析,筛选出差异表达的基因。结果：在所检测的人普通型软骨肉瘤与去分化软骨肉瘤组织中,31个基因有表达异常,其中高表达14条,低表达17条。结论：去分化软骨肉瘤与普通型软骨肉瘤间基因表达差异是明显的,这些差异基因主要分布于涉及TGT信号途径、Wnt信号途径、IHH/PThRP轴以及凋亡机制等多方面,且有些基因的作用目前尚不清楚。     

Rb-loss is associated with high malignancy in chondrosarcoma

Loss of function of the human retinoblastoma gene (Rb) is a frequent genetic abnormality in human malignancies and causes a disturbance in the cell cycle and loss of normal proliferation and differentiation. We studied the loss of heterozygosity (LOH) of the Rb gene in 31 formalin-fixed, paraffin-embedded cartilaginous tumors using polymerase chain reaction. The tumors were subdivided into 8 cases of dedifferentiated (DD) chondrosarcoma, 17 cases of conventional chondrosarcoma (nine grade 1, seven grade 2 and one grade 3), 4 enchondromas and 2 chondroblastomas. Both components of DD chondrosarcoma, the low-grade and anaplastic components, were separated by a microdissection approach. The genetic data were correlated with the expression of the Rb protein examined by Rb immunohistochemistry. We found Rb-LOH in one grade 3 chondrosarcoma, and in the anaplastic component in 7 of 8 cases of DD chondrosarcoma (89% of all high-grade chondrosarcomas). All tumors with Rb-LOH were immunohistochemically Rb-negative. The only case of DD chondrosarcoma negative for Rb-LOH in both components of the tumor also showed weak expression of the Rb protein in the anaplastic component. All benign cartilaginous tumors, low-grade chondrosarcomas and low-grade tumor components of DD chondrosarcomas were negative regarding Rb-LOH but positive in Rb immunohistostaining. We concluded that Rb-LOH predominantly occurs in high-grade chondrosarcomas. However, it is not a marker for identifying low-grade tumors with a tendency towards progression or local recurrence.     

Clinical features and treatment outcomes of dedifferentiated and grade 3 chondrosarcoma: A multi‐institutional study

Chondrosarcoma is the second most common primary malignant bone tumor. In this multicenter study, we sought to evaluate the disease‐specific survival (DSS) and disease‐free survival (DFS), and prognostic factors in patients with dedifferentiated chondrosarcoma (DDCS) or grade 3 chondrosarcoma (G3CS) in Japan. We retrospectively investigated the treatment outcomes and prognostic factors in 62 patients with DDCS and 19 patients with G3CS at 15 institutions participating in the Japanese Musculoskeletal Oncology Group. We also clarified significant clinicopathological factors for oncological outcomes. In surgery for primary lesions aimed at cure, a histologically negative margin (R0) was obtained in 93% (14/15) of patients with G3CS and 100% (49/49) of patients with DDCS. The 5‐year DSS was 18.5% in patients with DDCS and 41.7% in patients with G3CS (p = 0.13). Local control was obtained in 80% (12/15) and 79.6% (39/49) of patients with G3CS and DDCS in the primary lesion after surgery with a wide surgical margin, respectively. In multivariate analysis, stage and no treatment/palliative treatment for the primary lesion were independent prognostic factors for DSS of DDCS, and age and no treatment/palliative treatment for DSS of G3CS. The 5‐year DFS rate was 22.8% in 26 patients with DDCS who did not receive adjuvant chemotherapy, and 21.4% in 14 patients who received adjuvant chemotherapy. The prognosis of DDCS remains poor, although R0 resection was carried out in most cases. Effective and/or intensive chemotherapeutic regimens or agents should be considered or developed for patients with high‐grade chondrosarcoma, particularly for those with DDCS.     

Genomic alterations in low-grade,anaplastic astrocytomas and glioblastomas

To extend our understanding of potential stepwise genetic alterations that may underlie tumor progression from low-grade astrocytomas to glioblastomas, histopathologic and comparative genomic hybridization analyses were performed on tumor specimens from 68 primary lesions, including 40 glioblastomas, 10 anaplastic and 18 low-grade astrocytomas. The number of aberrations per case increased towards the higher grade tumors (grade II: 1.66±1.49; grade III: 2.80±1.68; grade IV: 3.02±1.07; F=6.955, p=0.002). A gain of 7/7q was common and the most frequently seen aberration in low-grade astrocytomas, whereas loss of 10q was the most frequently seen anomaly in anaplastic astrocytomas and glioblastomas. Chromosome 7p amplification was only detected in glioblastomas. Chromosome 10/10q deletion and combination of lp, 19q and 17p deletions were specific to high-grade astrocytic tumors. Sequences of chromosome 7 and 10 seem to have pivotal roles in the biology of human gliomas. The genomic copy deletions of chromosomes lp and 19q might provide an alternative mechanism in the genesis of astrocytomas.     

Deletion mapping of chromosome 19 in human gliomas

There is evidence that a putative glioma tumor suppressor locus resides on the long arm of chromosome 19. We present data on 161 gliomas from IS6 patients, which were studied by microsatellite analysis for loss of heterozygosity (LOH) on chromosome 19. Eight loci on the long arm and 2 loci on the short arm of chromosome IV were examined. LOH on I9qwas observed in 3/19 astrocytomas (WHO grade II), 12/27 anaplastic astrocytomas (WHO grade III), 16/76 cases of glioblastoma multiforme WHO (grade IV), 4/9 oligodendrogliomas (WHO grade II), 3/5 anaplastic oligodendrogliomas (WHO grade III), 5/9 mixed oligo-astrocytomas (WHO grade II) and 8/10 anaplastic oligo-astrocytomas (WHO grade III). While 31 of the tumors with LOH on chromosomal arm I9q exhibited allelic loss at every informative locus, 20 tumors showed terminal or interstitial deletions. In contrast to astrocytomas and glioblastomas, tumors with an oligodendroglial component had predominantly lost the entire long arm of chromosome 19. The common region of overlap in gliomas was located on 19q 13.2-q 13.4 between the markers D 19S 178 and D 19S 180. Our data confirm the involvement of a putative tumor suppressor gene on chromosomal arm 19q in gliomas and assign this gene to 19q 13.2-q 13.4.     

p53 expression in dedifferentiated chondrosarcoma

Background. p53 acts as a tumor suppressor gene because of to its negative control of the cell cycle and its central role in programmed cell death. It frequently is mutated, as observed in a variety of human neoplasms. The mutations inhibit tumor-suppressor activities of p53, which may gain a new function of tumor promotion. In this study, p53 was investigated in various components of dedifferentiated chondrosarcoma and correlated with their proliferative activities. Methods. Immunohistochemical assays for p53, Ki-67, and proliferating cell nuclear antigen (PCNA) were used in a series of eight dedifferentiated chondrosarcomas of bone. The cartilaginous component was low grade (Grade I-II) in five cases. It was predominantly low grade with foci of a high grade (Grade III) chondrosarcoma in the remaining three cases. The noncartilaginous (de-differentiated) high grade component consisted of malignant fibrous histiocytoma in five cases and osteosarcoma in three. Results. Regardless of the histological type, diffuse strong nuclear staining for p53 occurred in the high grade noncartilaginous component of all eight of the tumors. The low grade cartilaginous component of six cases was negative for p53, with focal weak staining in the two remaining cases. The high grade cartilaginous component showed strong positive staining for this protein in all three cases. Ki-67 and PCNA expression were similar to that of p53. Conclusions. The percentage of p53 positive staining roughly was parallel to the proliferating fraction of cells in various components of dedifferentiated chondrosarcoma. Moreover, p53 overexpression was consistently present in the high grade noncartilaginous (dedifferentiated) component of the tumor and was accompanied by increased proliferative activity. Cancer 1995; 76:223-7.     

Prognostic variables in oligodendroglial tumors: a single-institution study of 95 cases

We analyzed the relationships among clinical variables, histology, 1p/19q status, and outcome in 95 patients with oligodendroglial tumors. The study enrolled adult patients who underwent first-time surgery for a supratentorial oligodendroglial tumor at Oslo University Hospital, Rikshospitalet. Tumors were: 27 oligodendrogliomas, WHO grade II; 32 oligoastrocytomas, WHO grade II; 16 anaplastic oligodendrogliomas, WHO grade III; 14 anaplastic oligoastrocytomas, WHO grade III; and 6 glioblastomas with a major oligodendroglial component, WHO grade IV. The clinical files were reviewed. Three neuropathologists evaluated the histological slides independently. Loss-of-heterozygosity analysis for 1p and 19q was performed by PCR. Favorable prognostic factors from univariate analyses included seizures as presenting symptom, female sex, location in the frontal lobe, low WHO grade, classic histology, absence of gemistocytic cells, and combined 1p/19q loss. Solitary 19q loss was a negative prognostic marker. 1p/19q status was of prognostic significance in both tumors with classic and nonclassic oligodendroglial histology. In the multivariate analysis, WHO grade II (P< .001), frontal tumor location (P= .002), and combined 1p/19q loss (P< .001) remained favorable prognostic variables. Our results suggest that tumor location, WHO grade, and 1p/19q status are important independent variables associated with survival in oligodendroglial tumors. The study suggests that solitary 19q loss is a negative prognostic variable and that 1p/19q loss is associated with prolonged survival also in oligodendroglial tumors without classic histology.     

Genetic and pathologic evolution of early secondary gliosarcoma

Gliosarcoma is a subset of glioblastoma with glial and mesenchymal components. True secondary gliosarcomas (i.e. progressing from lower-grade precursors) in the absence of radiation therapy are very rare. We report the unique case of a 61-year-old male who developed a fibrillary astrocytoma (WHO grade II). In the absence of adjuvant therapy the tumor recurred 3 years later as a gliosarcoma comprising an infiltrating glial component and a curious, early high-grade sarcomatous component surrounding intratumoral vessels. DNA was extracted from formalin fixed paraffin-embedded tissues from the precursor low-grade glioma and from the glioma and sarcomatous components at progression. Samples were hybridized separately to a 300 k Illumina SNP array. IDH1(R132H) mutant protein immunohistochemistry was positive in all tissue components. Alterations identified in all samples included dup(1)(q21q41), del(1)(q41qter), del(2)(q31.1), del(2)(q36.3qter), del(4)(q35.1qter), dup(7)(q22.2q36.3), del(7)(q36.3qter), del(9)(p21.3pter), dup(10)(p13pter), del(10)(q26.13q26.3), dup(17) (q12qter), and copy neutral LOH(20)(p11.23p11.21). The recurrent tumor had additional alterations, including del(3)(p21.31q13.31), del(18)(q21.2qter), and a homozygous del(9)(p21.3)(CDKN2A locus) and the sarcoma component had, in addition, del(4)(p14pter), del(6)(q12qter), del(11)(q24.3qter), and del(16)(p11.2pter). In conclusion, unique copy number alterations were identified during tumor progression from a low-grade glioma to gliosarcoma. A subset of alterations developed specifically in the sarcomatous component.     

Phase II trial of merbarone in patients with malignant brain tumors

The standard treatment for patients with primary malignant glioma includes surgical resection, radiotherapy, and nitrosourea. Despite this multimodality approach, adults with newly diagnosed glioblastoma multiforme (GBM) and high-grade astrocytoma have a median survival duration of 50 weeks and 150 weeks respectively. Chemotherapy has had a limited impact on the survival of these patients. Merbarone (5-phenylcarboxamide-2-thiobarbituric acid) is a nonsedating derivative of barbituric acid that crosses the blood brain barrier. Antitumor activity of merbarone has been described against L1210, B16 melanoma cell line and the M5076 sarcoma cells in phase I studies. Merbarone inhibits DNA synthesis and tumor growth by inducing single strand breaks in DNA. It also inhibits RNA and protein synthesis. We evaluated merbarone in a phase II trial in patients (pts) with recurrent or refractory GBM (7 pts) and high grade anaplastic astrocytoma (7 pts). Fourteen patients (nine males, five females) were treated with merbarone at a dose of 1000 mg per m² per day by continuous intravenous infusion for 5 days every 3 weeks. Every patient received at least two cycles of treatment No complete or partial responses were observed, although one patient had stable disease lasting 20 weeks. Our conclusion is that merbarone is ineffective against GBM and high-grade anaplastic astrocytoma at the dose and schedule in which it was administered in this trial.     

Morphologic and molecular genetic aspects of oligodendroglial neoplasms

Morphologic criteria for diagnosing oligodendrogliomas and for classifying them as well-differentiated (World Health Organization grade II) and anaplastic (World Health Organization grade III) are well recognized. Nevertheless, applying these guidelines to specific cases often reveals discrepancies among different observers. In addition, whether a given tumor also contains an astrocytic component may be debatable. Loss of heterozygosity studies have demonstrated that oligodendroglial neoplasms have a high incidence of loss of the 1p and 19q chromosomal arms. Although loss of heterozygosity for portions of 19q are sometimes seen in astrocytic neoplasms, these tumors seldom show complete loss of 19q accompanied by loss of 1p. Loss of 9p or homozygous deletion of the CDKN2 gene or both are associated with anaplastic oligodendrogliomas, whereas loss of 17p or TP53 gene mutations or both are frequent in astrocytomas, but rare in oligodendrogliomas. These observations suggest that molecular genetic parameters could provide an objective, reproducible framework for classifying oligodendroglial neoplasms.     

Primary malignant bone neoplasm: a case report of dedifferentiated chondrosarcoma in the rib and review of the literature

Dedifferentiated chondrosarcoma (DDCS) is a rare but highly malignant primary bone neoplasm, which is resistant to radiotherapy and chemotherapy. There remains uncertainly as to the best treatment of this disease and how to improve its prognosis. In this paper we reported a case of DDCS and reviewed the related literatures in order to provide references to throw a light on the histogenesis, diagnosis and therapy of this disease.