胆囊腺瘤p53蛋白检测

1　材料与方法1 1　临床资料　标本为武警新疆总队医院 1992年 6月至1999年 12月间手术或腹腔镜摘除的胆囊腺瘤 32例 ,其中男性 2 0例 ,女性 12例 ,年龄 34～ 6 8岁 ,平均 5 7岁。并选出2 0例胆固醇性及炎性息肉做对照。1 2　免疫组化染色　标本经 10 %福尔马林液固定 ,石蜡包埋 ,5 μｍ厚切片 2张 ,分别做ＨＥ染色和ＡＢＣ法 ｐ5 3免疫组化染色。鼠抗人单克隆抗体ＤＯ 7,丹麦Ｄａｋｏ公司产品。按照ＡＢＣ试剂盒 (美国Ｖｅｃｔｏｒ公司 )操作程序进行 ,ＤＡＢ显色 ,苏木精复染。阴性对照以ＰＢＳ代一抗 ,阳性对照用已知ｐ5 3蛋白阳性的胃癌…     

p53蛋白检测方法的探讨

ｐ５３蛋白检测方法的探讨吴霞，黄晓南，杨德忠，邓永江，黄京丽ｐ５３抑癌基因的产物位于细胞核，是一种由３９３个氨基酸组成的含磷蛋白。正常的ｐ５３基因（即野生型）是一种抑癌基因，半衰期很短，而且极不稳定，一般方法很难检测，但突变的ｐ５３蛋白稳定性增加，半...     

乳腺癌中p53基因缺失及p53蛋白的异常表达

乳腺癌中ｐ５３基因缺失及ｐ５３蛋白的异常表达郭文斌张嘉庆杨德启乔新民张庆广阚秀虞有智一、材料与方法１．材料来源：４７例乳腺癌及正常腺体均来自我院乳腺中心１９９４～１９９６年间手术切除标本。全部病例均有病理学诊断。２．Ｓｏｕｔｈｅｒｎ杂交：用酚、氯仿／...     

p53和p21蛋白在恶性间皮瘤的表达

一、材料和方法１－集自１９８７年１月至１９９７年１２月间我院和广州地区多所医院的恶性间皮瘤石蜡包埋的组织标本４５例；所有病例标本重新进行组织学复诊、分型，并结合胸片，ＣＴ和手术所见进行大体分型及对其中的３６例进行随访。另外同时收集了良性间皮瘤８例，增生间皮细胞４２例作对照。２－方法：石蜡切片按常规处理，使用ＤＡＫＯ公司的免疫试剂盒及一抗，采用Ｅｎｖｉｓｉｏｎ二步法进行免疫组化染色。工作浓度ｐ５３为１∶１００；ｐ２１为１∶５０；ＤＡＢ显色。每次实验均设阳性及阴性对照，及用ＴＢＳ替代一抗的空白对照。…     

非霍奇金淋巴瘤与p53蛋白表达的关系

目的 :探讨非霍奇金淋巴瘤 (NHL)与 p5 3蛋白表达的关系。 方法 :用免疫组化S P法检测 10 2例 (低度恶性 2 3例 ,中度恶性 36例 ,高度恶性 4 3例 )NHLp5 3蛋白表达 ,根据 p5 3蛋白阳性细胞百分率将其表达水平分为 4级 :0级 (阴性 ) ,1级 (1%～ 2 5 % ) ,2级 (2 6 %～ 5 0 % ) ,3级 (>5 0 % )。结果 :低度恶性组 2 0 / 2 3(87% )p5 3表达为 0级 ,中度恶性组 31/ 36 (86 1% )表达为 1级 ,高度恶性组 33/ 4 3(76 7% )表达为 2～ 3级。 2 5例随访 7～ 6 8个月 ,p5 30～ 1级NHL完全缓解率 (CRR ,11/ 14 )高于p5 32～ 3级NHLCRR (1/ 11,P <0 0 1) ,前者生存率 (13/ 14 )高于后者 (3/ 11,P <0 0 1)。NHLp5 3蛋白表达水平与其恶性度密切相关 (P <0 0 1)。结论 :p5 3蛋白表达阳性细胞百分率是判断NHL恶性度、疗效及预后较可靠的参数。肿瘤性p5 3蛋白表达检测对中高度恶性NHL的诊断有参考价值     

甲状腺肿瘤p14ARF、p53及FHIT蛋白的表达

目的探讨p14^ARF、p53及FHIT蛋白在甲状腺肿瘤组织中的表达及其意义。方法采用免疫组化S-P法检测20例甲状腺腺瘤和28例甲状腺癌组织中p14^ARF、p53及FHIT蛋白的表达。结果p14^ARF、p53及FHIT蛋白在甲状腺腺瘤和甲状腺癌中阳性率分别为90％、35．7％、15％和75％、90％、7．14％,3种蛋白在甲状腺腺瘤及甲状腺癌的表达差异均有显著性（P〈0．05）。p14^ARF、p53及FHIT蛋白的表达在甲状腺滤泡癌（FTC）与腺瘤之间,乳头状癌（PTC）与腺瘤之间差异均有显著性（P〈0．05）,p53及FHIT的表达在髓样癌（MTC）与腺瘤间差异也有显著性（P〈0．05）。p14^ARF、p53及FHIT蛋白的表达与甲状腺肿瘤的恶性进程有关,与患者年龄、性别及淋巴结转移无关。另外p14^ARF与FHIT蛋白的表达呈正相关,并且它们与p53的表达均呈负相关。结论肿瘤抑制蛋白p14^ARF和FHIT的缺失以及p53蛋白的高表达可能是甲状腺肿瘤发生的重要原因之一。联合检测p14^ARF、p53及FHIT蛋白有助于区分甲状腺腺瘤和甲状腺滤泡癌。     

鼻咽癌癌旁粘膜的p53蛋白免疫组织化学观察

我们曾报道过鼻咽癌（ＮＰＣ）组织中ｐ５３蛋白积聚的生物学特征和临床意义［１］，但是ｐ５３基因的突变发生在ＮＰＣ发生发展中的哪个阶段尚不清楚，为此我们以ＮＰＣ癌旁粘膜组织为材料，用免疫组化和图像定量分析的方法，观察癌旁粘膜上皮ｐ５３蛋白积聚的情况，以判...     

结直肠癌mdm2,p21和p53蛋白的表达

一、材料与方法1.标本 :89份石蜡组织取自 1991～ 1994年期间本院结直肠腺癌手术切除标本 ,术后随访 3～ 6年。其中高分化腺癌 30例 ,中分化腺癌 44例 ,低分化腺癌 15例。按Ｄｕｋｅｓ临床分期 ,无Ａ期病例 ,Ｂ期 42例 ,Ｃ期 37例 ,Ｄ期 10例。纤维结肠镜及病理检查未发现异常的结直肠粘膜活检标本 12例做对照。组织均常规甲醛溶液固定 ,石蜡包埋 ,连续切片 ,分别作常规ＨＥ染色 ,ｍｄｍ2、ｐ2 1、ｐ5 3蛋白免疫组化染色。2 .免疫组化染色 :采用ＳＰ法 ,鼠抗ｍｄｍ2 (ＳＭｐ14)、ｐ5 3(Ｄｏ 1)、ｐ2 1(187)为ＳａｎｔａＣｒｕｚ公司试剂 ,Ｓ…     

子宫内膜癌PTEN及p53蛋白的表达

在子宫内膜癌的研究中已经确定了的、较为常见的分子生物学改变有ｐ5 3突变、Ｋ ｒａｓ突变和微卫星不稳定性 ,大约各占子宫内膜癌的 2 0 %～ 3 0 % [1] 。已发现子宫内膜癌中ＰＴＥＮ抑癌基因 (ｐｈｏｓｐｈａｔａｓｅａｎｄｔｅｎｓｉｎｈｏｍｏｌｏｇｄｅｌｅｔｅｄｏｎｃｈｒｏｍｏｓｏｍｅｔｅｎ)突变率为 3 2 %～ 5 5 % ,是迄今为止子宫内膜癌中所发现的突变率最高的基因改变[2 4 ] 。我们拟通过观察子宫内膜癌ＰＴＥＮ及ｐ5 3蛋白表达的情况来探讨其与子宫内膜癌的发生、发展和预后的关系。一、材料和方法80例子宫内膜癌标本选自北…     

p53的两种结合蛋白:53BP1和53BP2

p53基因是一种广谱的肿瘤抑制基因,其产物p53为一多功能的转录调节因子,可以发挥调节细胞生长、细胞凋亡和DNA修复的作用。在人类肿瘤已发现多种p53基因的点突变,但突变并不是p53蛋白质丧失功能的唯一途径。胞内蛋白可能影响其活性和功能。53BP1和53BP2是p53在胞浆中的两种结合蛋白。本文阐述了有关这两种蛋白质的研究进展。     

p53/Rb细胞转导通路相关基因和蛋白在衰老大鼠睾丸组织的表达

目的 通过检测p53、Rb、p16、p19、p21在衰老大鼠睾丸组织的表达,探讨p53/Rb细胞转导通路相关基因和蛋白在睾丸组织衰老中的意义. 方法 选用8周龄雄性SD大鼠30只,体重180～220g,随机分为正常对照组和模型组,每组15只.模型组大鼠采用D-半乳糖连续腹腔注射建立亚急性衰老大鼠模型.采用RT-PCR方法 检测p53、Rb基因在大鼠睾丸组织的表达;Western blotting法检测大鼠睾丸磷酸化Rb、p16、p19、p21蛋白的表达.结果 模型组睾丸组织p53、Rb基因表达明显高于正常对照组(P<0.01);模型组睾丸组织与正常组比较:p16、p19、p21蛋白的表达明显升高,而磷酸Rb蛋白表达显著下降(P<0.01). 结论 Rb/p53细胞转导通路相关基因及蛋白在衰老大鼠睾丸组织发生明显改变,Rb/p53细胞转导通路阻滞可能在睾丸组织的衰老过程中起着一定作用.     

PROGNOSTIC SIGNIFICANCE OF THE CO-EXPRESSION OF p53 AND c-erbB-2 PROTEINS IN BREAST CANCER

The immunohistochemical expression of p53 and c-erbB-2 gene proteins was examined in a series of 130 breast adenocarcinomas. This study intended to investigate whether the frequency of the altered expression of the tumour suppressor gene p53 and the overexpression of the oncogene c-erbB-2 in breast cancer tissue cells correlated with other variables known to affect the biological behaviour of these tumours and the overall survival of the patients (median follow-up time: 6 years). The expression of p53 protein and c-erbB-2 gene product was evaluated immunohistochemically. Expression of p53 protein was detected in 30 (23 per cent) of the neoplasms examined, while 26 (20 per cent) out of the 130 cases demonstrated positive c-erbB-2 immunoreactivity. There was a statistically significant association between p53 protein expression and primary tumour size, lymph node involvement, and oestrogen receptor positivity. The incidence of c-erbB-2 positivity was significantly correlated with high tumour grade, axillary node invasion, large tumour size, and the absence of steroid receptors. p53 immuno-expression was clearly associated with c-erbB-2 protein overexpression. Concomitant p53 and c-erbB-2 positive immunolabelling, which emerged in 14 out of the 130 cases (10·7 per cent), was clearly associated with high grade, large size, positive nodal status, ductal infiltrating (NOS) histological type, and low values of progesterone receptors. Overall survival of patients was not significantly related to the immunoreactivity of either p53 or c-erbB-2 considered separately, whereas there was a clearly significant trend to worse overall prognosis in cancers with double p53/c-erbB-2 positive phenotype. The simultaneous immunodetection of p53/c-erbB-2 appears to have greater negative prognostic relevance than their separate expression.     

p53 Protein expression in laryngeal squamous cell carcinomas bearing wild-type and mutated p53 gene

We performed an immunohistochemical analysis to investigate the expression of p53 protein in a panel of 18 laryngeal squamous cell carcinomas, 15 primary tumours and three in relapse, previously analysed by us for the presence of p53 gene mutations. Dysplastic and/or normal surrounding mucosa was evaluated in 15 different tumours. The results of our study are the following: (1) expression of p53 protein was observed in one out of five tumours positive for p53 gene mutations (20%) and in 10 out of 13 (80%) negative cases; (2), p53 protein over-expression was frequently observed in normal and/or dysplastic mucosa surrounding either wild-type (7/11) or mutated p53 tumours (2/4); (3), p53 immunoreactive cells showed a pattern of distribution in normal and mildly/moderately dysplastic mucosa (basal layers), different from that in severely dysplastic mucosa (whole thickness). These data further support the hypothesis that p53 protein over-expression may be a marker of the earliest phases of multistep tumorigenesis in laryngeal squamous cell carcinoma.     

脂质体介导的p53基因对肝癌细胞生长的抑制作用

目的 观察外源野生型ｐ５３基因在肝癌基因治疗方面的可行性。方法 将载有人野生型ｐ５３－ｃＤＮＡ的真核表达质粒ｐ５３－ｐｃＤＮＡ３,用阳离子脂质体介导转染人肝癌细胞系ＨｅｐＧ２,用流式细胞仪检测ｐ５３－ｐｃＤＮＡ３对ＨｅｐＧ２细胞生长的影响。结果 通过观察细胞生长曲线与流式细胞仪检测细胞周期和细胞的凋亡指数发现,ＨｅｐＧ２细胞生长受到明显的抑制。结论 脂质体介导的ｐ５３基因可在Ｈ细胞中表达,且明显抑     

端粒重复因子2与p53的体外结合

目的 通过分析端粒主要结合蛋白端粒重复因子2(TRF2)与p53的体外结合,探讨p53通过端粒途径调节细胞增殖、衰老和凋亡的分子机制。方法 4种不同的p53-谷胱甘肽S转移酶(glutathione S-transferase,GST)融合蛋白和GST经大肠杆菌表达、谷胱甘肽-Sepharose^TM 4B纯化,其中的人重组p53包括野生型(1～393)、C端缺失体p53 N5(2～293)、N端缺失体p53 N5(95～393)、第175位氨基酸突变体(R→H)。将各纯化蛋白和人乳腺癌细胞MCF-7的细胞蛋白进行体外结合反应(pull down),Western blot检测反应物中p53和TRF2的结合。结果 纯化的GST和p53-GST融合蛋白纯度均在90％以上,且相对分子质量与预计的完全一致。TRF2的Western blot显示：野生型p53和p53-R 175H均能沉淀MCF-7中的TRF2,且结合力相似,而单独的GST则无沉淀TRF2的作用。与野生型p53和p53 R175H相比,p53 2C与TRF2的结合力相对增加,p53 N5与TRF2的结合力相对大大减弱。结论 p53和TRF2可以进行直接而特异的体外结合,且其结合部位在p53的C端(293～393)。p53和TRF2的C端依赖性结合可能与端粒动态变化所诱导的细胞活动有关。     

p53 OVEREXPRESSION AND HUMAN PAPILLOMAVIRUS INFECTION IN TRANSITIONAL CELL CARCINOMA OF THE URINARY BLADDER: CORRELATION WITH HISTOLOGICAL PARAMETERS

Seventy-nine transitional cell carcinomas (TCCs) of the urinary bladder (25 grade 1, 22 grade 2, and 32 grade 3 tumours) were examined for p53 overexpression by immunohistochemistry with a monoclonal antibody and for human papillomavirus (HPV) infection by the polymerase chain reaction (PCR). Positive immunostaining for p53 was detected in 40·5 per cent of the cases; the percentage of positive cases was significantly lower in low-grade (G1 and G2) TCCs than in high-grade (G3) tumours (10·6 per cent vs. 84·4 per cent; P <0·0001). The overall rate of HPV infection was 32·9 per cent; 20·3 per cent of the cases were positive for HPV 16, 3·8 per cent for HPV 18, and 8·9 per cent for both. Consensus primers as well as type-specific primers for HPV types 6, 11, and 33 failed to detect any additional case with HPV infection. The prevalence of HPV 16 and/or HPV 18 infection was significantly higher in low-grade than in high-grade tumours (44·7 per cent vs. 15·6 per cent; P =0·0061). p53-positive cases were more common among papillary, deeply infiltrating tumours, and HPV-positive cases among papillary, non-infiltrating lesions. According to these data, p53 overexpression and HPV 16/18 infection are common findings in bladder TCC and there appears to be an inverse correlation of p53 overexpression and of HPV infection with tumour aggressiveness. The possibility of different molecular pathways in superficial low-grade and in invasive high-grade tumours is suggested.     

外源性p53基因对人胃粘膜上皮细胞系GES－1的生物学效应

为了解ｐ５３基因在细胞癌变过程中的作用，分别将野生型和突变型的ｐ５３基因导入体外培养的人胃粘膜上皮细胞系ＧＥＳ－１中，进一步比较，分析转染ｐ５３基因对ＧＥＳ－１细胞的生物学效应发现：（１）外源ｐ５３基因改变了ＧＥＳ－１细胞的形态──转染野生型ｐ５３基因使得ＧＥＳ－１出现一些细胞分化的迹象，在其胞浆有明显的空泡，空泡周围有微管的环绕；（２）ｐ５３基因的转染改变了ＧＥＳ－１生长状态──野生型的ｐ５３基因转染的细胞的克隆形成率低于突变型ｐ５３基因以及空载体ｎｅｏ转染的细胞。实验还发现突变型ｐ５３基因改变了细胞生长习性，使细胞有选择生长优势，对细胞恶性度有促进作用。     

Cell-type-specific expression of p53 and p21 in giant cell arteritis

The aim of the present study was to investigate the expression of TP53 (p53) and CDKN1A (CIP1; p21) in the arterial wall in giant cell arteritis (GCA). Cross-sections from 18 temporal artery biopsies displaying GCA and 8 control arteries were double-stained with monoclonal antibody directed at p53 or p21 on the one hand and alpha-smooth muscle actin, CD68 (macrophage) or CD3 (T-cell) on the other. Nuclear p53 was expressed in CD68-positive cells and smooth muscle cells in 16 of the 18 inflamed arteries. P21-positive nuclei were found in CD68-positive cells in 14 biopsies and in smooth muscle cells in all the specimens. All p53-positive giant cells also contained p21-positive nuclei. In the giant cells, immunopositive nuclei were mixed with negative ones. CD3-positive T-cells did not express p53 or p21. Only one p53-positive smooth muscle cell nucleus was found in the non-GCA controls and, compared with GCA, p21 expression was noted in few smooth muscle nuclei. The presence of p53 and p21 in the same types of cell in GCA indicates that the former protein is functional; p21 expression is induced by wild-type, functional p53 but not by its mutant form. The current observations suggest cellular stress in GCA, the nature of which requires further investigation.