p53在涎腺良恶性多形性腺瘤中的表达及其意义

p53基因是迄今发现与人类肿瘤相关性最高的肿瘤抑制基因。本文对涎腺多形性腺瘤和癌在多形性腺癌中的p53蛋白进行检测，旨在明确p53基因在这二种良恶性肿瘤中的堆积特性及其临床意义。1材料与方法选择涎腺肿瘤标本39例，其中多形性腺瘤20例，癌在多形性腺瘤中19例，常规固定、包理，连续切片2张，厚4μm，1张HE染色，另1张用ABC法做免疫组化染色（一抗为鼠抗人基因重组p53蛋白抗体）2结果全部病例的组织切片背景清晰，免疫反应定位于肿瘤细胞核内，呈棕黄色或棕褐色。19例涎腺癌在多形性腺瘤中有4例阳性，占21％（图1）。其中1例恶性成份…     

涎腺良性多形性腺瘤和恶性多形性腺癌中c—erbB—2和bcl—2蛋白？…

目的：为了探讨细胞增殖和细胞凋亡的调控基因在涎腺肿瘤的发生发展及恶性转化中的作用。方法：采用免疫组化方法对５例瘤旁腺体，２１例良性多形性腺瘤和２４例恶性多形性腺瘤中ｃ－ｅｒｂＢ－２和ｂｃｌ－２蛋白产物的表达进行形态学观察。     

脑膜瘤细胞凋亡及凋亡相关基因bcl—2、p53蛋白的表达

目的 探讨脑膜瘤自发性细胞凋亡相关基因bcl-2、p53蛋白在脑膜瘤中的表达及其意义。方法 采用TUNEL法和免疫组化S－P法检测40例脑膜瘤中细胞凋亡及凋亡相关基因bcl-2,p53蛋白的表达。结果 恶性和非典型脑膜瘤中的细胞凋亡指数（AI）明显高于良性脑膜瘤（P＜0．05）,良性脑膜瘤不同亚型之间bcl-2,p53表达无显著性差异（P＞0．05）;p53蛋白表达随脑膜瘤恶性程度增高而增强,具有显著性差异（P＜0．05）,p53阳性与阴性的脑膜瘤中的细胞凋亡指数无显著性差异;bcl-2蛋白表达与脑膜瘤细胞凋亡呈负相关。结论 恶性及非典型脑膜瘤中自发性细胞凋亡增多;凋亡是脑膜瘤恶性或不典型的1个重要的生物学行为;p53基因在脑膜瘤的恶变过程中发生重要作用,与脑膜瘤的恶性进展密切相关;bcl-2基因在脑膜瘤中可抑制细胞凋亡。     

p16抑制基因在腮腺多形性腺瘤恶性增殖中的作用

涎腺多形性腺瘤是比较常见的肿瘤．约占涎腺肿瘤的50％。p16基因(MTS1，CDK4I，CDKN2)是细胞周期的调控者，可能是在肿瘤发生中起重要作用的肿瘤抑制基因．主要与其编码蛋白产物调控细胞增殖有关。本研究应用流式细胞仪检测了不同类型的腮腺多形性腺瘤细胞周期变化．同时利用计算机图像处理系统，对p16基因在腮腺多形性腺瘤中的蛋白表达进行相对定量，探讨p16基因在腮腺多形性腺瘤发生、发展及在恶性增殖中的作用．为涎腺肿瘤的深人研究、肿瘤患者的早期辅助诊断和判断预后提供新的观点和途径。     

Hp感染与胃癌及癌前病变中p53、bcl-2、c-myc基因表达关系的研究

目的：研究幽门螺杆菌（Hp)感染的胃癌及癌前病变患者胃粘膜p53,bcl-2,c-myc基因的表达，以探讨Hp在胃癌发生，发展过程中的作用机制，方法：应用免疫组化方法测定60例胃癌组织，18例异型增生，29例肠上皮化生，35例萎缩性胃炎，29例浅表性胃炎组织中Bcl-2,c-myc基因蛋白表达情况，PCR／SSCP测定p53基因第5－8外显子突变，Hp阳性由快速尿素酶法和HE染色确定，结果：胃癌组Hp阳性41例，阴性19例，在胃癌组织中p53,bcl-2,c-myc基因阳性表达分别为38例（63．3％），38例（63．3％），18例（30．0％），p53,c-myc基因在胃癌中阳性表达率显著高于异型增生（P＜0．05）,bcl-2在异型增生组织中的阳性表达率与胃癌，肠上皮化生相比无显著性差异（P＞05），p53,bcl-2,c-myc基因表达与胃癌分期，淋巴结转移均无显著性关系（P＞0．05），但bcl-2与胃癌类型，分化程度显著相关（P＜0．05），每一种病变类型Hp阳性组p53,bcl-2,c-myc基因蛋白的阳性表达率均高于Hp阴性组，两者之间有显著性差异（P＜0．05），结论：Hp感染者具有更多肿瘤生物学行为，可以引起抑癌基因p53第5－8外显子突变，癌基因c-myc，凋亡调节基因bcl-2表达增加,Hp可能为促癌剂，在胃癌的发生发展中起一定作用。     

涎腺良性多形性腺瘤和恶性多形性腺瘤中c-erbB-2和bcl-2蛋白产物的表达及其意义

目的：为了探讨细胞增殖和细胞凋亡的调控基因在涎腺肿瘤的发生发展及恶性转化中的作用。方法：采用免疫组化方法对５例瘤旁腺体，２１例良性多形性腺瘤和２４例恶性多形性腺瘤中ｃ－ｅｒｂＢ－２和ｂｃｌ－２蛋白产物的表达进行形态学观察。结果：ｃ－ｅｒｂＢ－２和ｂｃｌ－２在良性多形性腺瘤中均有过度表达，表达率分别为２９％和１９％，主要分布于腺管样结构的腺上皮细胞的胞浆中。在恶性多形性肿瘤中ｃ－ｅｒｂＢ－２的表达率为５８％，主要分布于腺癌细胞的胞膜；ｂｃｌ－２表达率为４６％，主要分布于腺癌细胞的核膜及部分胞浆中。５例瘤旁腺体内导管上皮细胞也存在ｃ－ｅｒｂＢ－２和ｂｃｌ－２的过度表达。结论：ｃ－ｅｒｂＢ－２蛋白产物的过度表达在涎腺多形性腺瘤的发生发展中起着重要作用，同时也表明细胞凋亡基因的调控失常也是其重要的发病因素之一     

骨肉瘤细胞凋亡相关基因的表达及其临床意义

目的 探讨反映骨肉瘤预后的蛋白标记物。方法 对骨肉瘤石蜡组织作免疫组化和Tunel染色,研究p53 c-myc和bcl-2基因的表达与肿瘤细胞凋亡指数(Al)的关系,及其与病理类型和患者颅后的关系。结果 p53、c-myc和bcl-2的表达水平与细胞凋亡指数呈负相关,与病理类型无相关关性。p53、c-myc、bcl-2和细胞凋亡指数与患者远期生存密切相关。结论 p53、c-myc和bcl-2的表达水平以及细胞凋亡指数可作为判断骨肉瘤恶性程度和预后的指标,可指导临床进行治疗。     

EB病毒、p53、bcl-2、c-myc和Rb蛋白在非霍奇金淋巴瘤发病及病情演化中的价值

目的：探讨EB病毒（EBV）与bcl-2、突变型p53、c-myc和Rb蛋白在非霍奇金淋巴瘤（NHL）发病及病情演化中的价值。方法：用免疫组化S-P法对140例NHL进行EBV、bcl-2、突变型p53、c-myc和Rb蛋白检测，并选择同期增生性淋巴结炎51例作为对照组。结果：EBV、p53、bcl-2和c-myc蛋白在NHL表达的阳性率和Rb蛋白表达的阴性率均高于对照组，两组比较差异有显著性（P〈0.05）；EBV与突变型p53、bcl-2、Rb、c-myc蛋白在NHL中表达无相关性（P〉0．05）：突变型p53、bcl-2在不同恶性程度NHL中的表达有显著性差异（P〈0．05）；EBV、突变型p53、bcl-2、Rb、c-myc蛋白在不同临床分期NHL中的表达有显著性差异（P〈0．05）。结论：EBV、突变型p53、bcl-2、和c-myc蛋白与NHL的发病存在正相关关系，Rb蛋白则为负相关关系；EBV和突变型p53、bcl-2、c-myc、Rb蛋白在NHL的发病中系相对独立的因素；突变型p53、bcl-2蛋白表达与NHL恶性程度成正相关关系；EBV、突变型p53、bcl-2、和c-myc蛋白与NHL病情恶化成正相关关系，Rb蛋白则为负相关关系。     

p16和bcl—2基因在膀胱癌中的表达及意义

目的：探讨p16抑癌基因和 bcl-2凋亡抑制基因与膀胱癌生物学行为的关系。方法：采用免疫组化和核酸原位杂交等方法检测67例膀胱癌中p16和bcl-2基因的表达。结果：67例膀胱癌p16蛋白表达阳性率52．23％（35／67），失表达率47．76（32／67），其中Ⅰ，级，Ⅱ级，Ⅲ级的肿瘤p16蛋白失表达率各为14．55％，60．53％，71．43％（P＜0．05），p 16 mRNA失表达率分别为20％，40％，80％，40％，60％，100％（P＜0．05），免疫组化和原位杂交结果一致（P＞0．05），结论：p16基因表达与细胞分化呈负相关，bcl-2基因表达与细胞分化呈正相关，提示p16基因的缺失，突变和高甲基化，bcl-2基因的异常表达在膀胱癌的发生和发展中起重要的推动作用。两者均可作为判断膀胱癌生物学行为的参考指标。     

p53及PCNA在涎腺多形性腺瘤组织中的表达及临床意义

目的 探讨p53及PCNA蛋白在涎腺良、恶性多形性腺瘤组织中的表达及其临床意义.方法 应用免疫组化S-P法,检测25例恶性多形性腺瘤(MPA)、35例良性多形性腺瘤(PA)、22例腮腺正常组织中 p53和PCNA蛋白表达.结果 p53及PCNA蛋白表达在恶性多形性腺瘤分别为32.00%(8/25)和56.00%(14/25),在良性多形性腺瘤组织中的阳性表达率分别为17.14%(6/35)和22.86%(8/35),在腮腺正常组织的阳性表达率分别为0%(0/22)和9.09%(2/22),p53、PCNA在良、恶性多形性腺瘤组织中的阳性表达率显著高于腮腺正常组织(P<0.05).结论 p53、PCNA蛋白的过度表达可能与良、恶性多形性腺瘤发生发展有关.     

Expression and mutation patterns of p53 in benign and malignant salivary gland tumors

The expression and mutation patterns of p53 were studied in a series of 68 benign pleomorphic adenomas and 237 malignant salivary gland tumors. p53 overexpression (nuclear staining exceeding 10%) was detected in 20% of the malignant salivary gland tumors, with the highest prevalence observed in polymorphous low grade adenocarcinoma, squamous cell carcinoma, and carcinoma ex pleomorphic adenoma and the lowest in adenoid cystic carcinoma and acinic cell carcinoma. In contrast, none of the 68 benign pleomorphic adenomas had nuclear staining exceeding 10%. SSCP and nucleotide sequence analysis of exons 4 to 9 of p53 in 19 malignant tumors revealed 9 mutations in 7 tumors. Our findings indicate that p53 may be a useful marker to help discriminate between benign and malignant salivary gland tumors.     

涎腺多形性腺瘤的倍体分布及与基因治疗的关系

目的:探讨涎腺多形性腺瘤的DNA含量及倍体分布与HSV-tk基因治疗的关系.方法:采用流式细胞仪测定50例涎腺良性多形性腺瘤DNA含量和倍体分布并结合临床资料进行分析;应用腺病毒介导的HSV-tk基因分别转染培养中良性及交界性多形性腺瘤细胞;RT-PCR方法检测基因表达;四唑蓝比色(MTI)法测定HSV-tk系统对二倍体及异倍体肿瘤细胞的杀伤作用.结果:10例正常涎腺组织皆为二倍体;涎腺良性多形性腺瘤异倍体率20.0%;30岁以下年龄,病程在2年以内及瘤体直径＞5cm者异倍体率明显增高(P＜0.05).二倍体肿瘤细胞转染HSV-tk/GCV基因后3、5天,细胞存活率分别为78.3%和37.7%.异倍体肿瘤细胞转染HSV-tk/GCV基因后3、5天,细胞存活率分别为27.6%和27.2%,二者细胞存活率有显著性差异(P＜0.05).结论:1)涎腺多形性腺瘤异倍体检出率与患者的年龄,病程及瘤体直径有密切关系;2)HSV-tk/GCV在治疗后的3、5天对异倍体肿瘤细胞杀伤作用明显高于二倍体肿瘤.     

Involvement of p53 mutation in the development of human salivary gland pleomorphic adenomas.

We examined the status of the p53 mutation, a putative tumor suppressor gene, as well as the expressions of myc and mos oncogene products in human salivary gland pleomorphic adenoma cells in culture derived from four individuals using techniques which enabled selective and favourable growths of tumor cells. Culture techniques empolyed in this study consisted of type I collagen gel-coated dishes and serum-free medium as substrates and growth medium, respectively. Cells grown under above conditions were subjected to the analyses of p53, myc and mos expression. When analyzed by both immunocytochemical staining and immunoblot, mutant forms of p53 specifically detected by PAb240 were observed in three of 4 cases. However, none of the 4 cases expressed myc and mos oncogene products. These results may imply a role for p53 mutation in the development of human salivary gland pleomorphic adenomas.     

Glucose transporter type 1 expression are associated with poor prognosis in patients with salivary gland tumors

The objective of this study was to investigate the expression of Glucose transporter type 1 (GLUT1) and its relation to clinicopathologic parameters in patients with salivary gland tumors. Tissue samples were 49 cases of malignant tumors, 38 cases of benign tumors, mainly pleomorphic adenoma and 10 cases of normal salivary glands. Immunohistochemically, GLUT1 Labeling Index of malignant tumor was significantly higher than benign tumor. In RT-PCR assay, GLUT1 mRNA level in malignant tumor were higher than that of normal tissue. In malignant tumors, expression of GLUT1 correlated significantly with tumor size (p=0.002) and distant metastasis (p=0.007). Cumulative survival rate of high expression group was significantly lower than that of low (p<0.001). In multivariable analysis, overall survival significantly correlated with lymph node metastasis (p=0.042) and GLUT1 expression (p=0.022). These results suggest that GLUT1 expression may provide useful prognostic information in patients with salivary gland carcinoma.     

Apoptosis and bcl-2 expression as predictors of survival in radiation-treated non-small-cell lung cancer

OBJECTIVES: We assessed the role of apoptosis and the expression of bcl-2, p53, and c-myc oncoproteins in pretreatment histologic specimens as a predictor of response to radiation therapy and survival in non-small-cell lung cancer (NSCLC) patients. METHODS: Pretreatment biopsy specimens of 68 patients with NSCLC (62 squamous cell carcinoma, 6 adenocarcinoma) were stained with hematoxylin and eosin. From 5 high-powered fields, the apoptotic index (AI) was calculated as the ratio of apoptotic tumor cells to the total number of tumor cells. Bcl-2, p53, and c-myc oncoprotein expression was detected by immunohistochemical staining. RESULTS: Twenty-nine cases showed partial or complete remission, whereas 39 showed no response. AI ranged from 0.2 to 12.0% (mean +/- SD; 4.3 +/- 2.6%, median 4.0%). There was no difference in AI between responders (4.0 +/- 2.3) and nonresponders (4.5 +/- 2.8, p > 0.05). However, in the responders, AI was correlated with the degree of change in tumor volume (r = 0.41, p < 0.05). In an analysis of 53 subjects who survived more than 1 month after the completion of radiation therapy, the patients with a higher AI (n = 27, MST = 22.8 m) survived longer than those with a lower AI (n = 26, MST = 9.2, log-rank, p = 0.03). Patients expressing bcl-2 had poorer survival (n = 22, MST = 6.0 m) than patients without bcl-2 (n = 31, 22.8 m, p < 0.003). According to multivariate analysis, three variables, bcl-2 expression, AI, and response to radiation, were independent prognostic factors for survival. CONCLUSION: A low level of spontaneous apoptosis and expression of apoptosis blocking bcl-2 protein in pretreatment histology predict a poor prognosis for radiation-treated NSCLC patients.     

Immunohistochemical study of neuron specific enolase expression in salivary-gland tumors

The immunoreactivity of monoclonal anti-neuron-specific enolase (MoAb NSE) on 10% formalin-fixed sections of normal human salivary glands and tumors were examined by the avidin-biotin-peroxidase complex (ABC) method. MoAb NSE staining of ductal and acinar cells of the normal salivary glands were, negative, and peripheral nerve fiber in the gland tissue showed strongly positive staining. In salivary gland tumors, pleomorphic adenoma (34 in total 68), myoepithelioma (15 in 35), monomorphic adenoma (7 in 10), adenolymphoma (15 in 23), mucoepidermoid carcinoma (3 in 16), acinic cell carcinoma (1 in 7), adenoid cystic carcinoma (12 in 20) and sialocarcinoma (4 in 15), stained focally for MoAb NSE staining. Frequency of occurrence for positive NSE activity varies among benign and malignant salivary gland tumors. In addition to MoAb NSE, S-100 protein and GFAP also demonstrated positive reactions in pleomorphic adenoma (Simpson et al Cancer 54: 1364-1369, 1984). The outer layer cells and/or peripheral cells of tubulo-ductal structure as well as modified myoepithelial cells and the cells of solid structure coexpressed with NSE, S-100 protein and GFAP. It is postulated that the salivary gland tumors particularly pleomorphic adenoma may be neuroectodermal in origin, arising from stem cells in intercalated duct segments.     

PCNA和Ki-67在涎腺良、恶性多形性腺瘤中的表达及意义

目的：研究涎腺良、恶性多形性腺瘤的临床病理特点,生物学行为及细胞增殖活性,为其临床诊断及预后评估提供依据。方法：对114例涎腺良、恶性多形性腺瘤进行回顾性分析,对其中20例多形性腺瘤、9例多形性腺瘤生长活跃型及9例恶性多形性腺瘤采用SABC和LSAB免疫组化法观察增殖细胞核抗原（PCNA）和增殖细胞核相关抗原（Ki-67）的表达和分布。结果：恶性多形性腺瘤的PCNA及Ki-67表达水平显著高于多形性腺瘤及多形性腺瘤生长活跃型,其PI值较后二者差异有显著性（P〈0．01）;良性多形性腺瘤的复发组PCNA和Ki-67的表达明显高于原发组,二者之间有显著性差异（P〈0．01）。结论：PCNA和Ki-67检测在判断多形性腺瘤恶性增殖方面有重要意义,对良性多形性腺瘤复发预测有重要的参考价值。     

乳腺癌及其癌前病变中细胞凋亡与p53、bcl-2蛋白的表达

目的：通过观察乳腺癌及其癌前病变中细胞凋亡调控基因ｐ５３、ｂｃｌ－２的表达,探讨细胞凋亡与凋亡调控基因在乳腺组织恶性转化进程中的作用。方法：利用ＤＮＡ缺口末端标记技术和免疫组织化学染色,原位观察３１例乳腺癌,２０例乳腺不典型增生和２０例乳腺单纯性增生中细胞凋亡和ｐ５３、ｂｃｌ－２蛋白的表达,以８例正常乳腺组织作为对照。结果：乳腺不典型增生和单纯性增生中细胞凋亡指数显著高于乳腺癌及正常乳腺组织（ｐ〉