肺癌中p53和p63 蛋白表达的高通量组织微阵研究

目的探讨p53和p63蛋白在肺癌中的表达及意义,并比较两者之间的表达关系.方法组织微阵(tissue microarray,TMA)技术和免疫组化S-P方法(immunohistochemistry,IHC).结果p53蛋白IHC染色中,TMA有价值的肿瘤标本为234(72.9%),总的表达率(+、++、+++)率为48.3%(113/234).随着肺鳞状细胞癌(squamous cell carcinoma,SCC)和腺癌(adenocarcinoma,Ad)分级升高,p53蛋白表达增强,与肺SCC及Ad病理分级有明显相关性(P＜0.05).p53蛋白表达与生存期呈明显负相关性(P＜0.05).p53蛋白表达与病理分型和临床分期等无明显相关性(P＞0.05).p53蛋白表达在肺癌原发肿瘤和转移淋巴结之间无明显差异(P＞0.05).p63蛋白IHC染色中,TMA有价值的肿瘤标本为264(82.2%),总的表达率(+、++、+++)为67.8%(179/264).p63蛋白在小细胞肺癌(small cell lung carcinoma,SCLC)表达最低16.0%(4/25),非小细胞肺癌(non-small cell lung carcinoma,NSCLC)与SCLC之间表达存在明显差异(P＜0.05).p63蛋白表达与病理组织分级、临床分期、生存期等无明显相关性(P＞0.05).p63蛋白表达在肺癌原发肿瘤和转移淋巴结之间无明显差异(P＞0.05).结论1.p53是肺癌的独立预后指标;2.p63蛋白在肺癌的不同病理类型的不同表达,说明肺癌的不同病理类型分子机理有所不同;3.p53蛋白和p63蛋白表达之间无明显相关性;4.TMA可以有效分析比较组织中肿瘤标记物的表达情况.     

非小细胞肺癌组织p53基因家族成员的不同表达及意义

目的探讨p53基因家族成员p53、p63和p73在非小细胞肺癌(NSCLC)中的不同表达及其临床意义.方法利用免疫组化S-P法在60例NSCLC和7例正常肺组织中检测p53、p63和p73蛋白的表达.结果在NSCLC中,p53、p63和p73蛋白的阳性表达率分别为61.67%(37/60)、80.00%(48/60)和73.33%(44/60);与正常肺组织相比,p53、p63和p73蛋白阳性表达率的差异均有显著性(P＜0.05).p53蛋白表达与肺癌细胞分化程度有密切关系(P=0.023),而与组织学类型、淋巴结转移和临床分期均无明显关系(P＞0.05).p63蛋白表达与肺癌组织学类型(P=0.001)和淋巴结转移(P=0.028)有密切关系,而与细胞分化程度和临床分期无明显关系(P＞0.05).p73蛋白表达与肺癌的临床病理特征均无明显关系(P＞0.05).在NSCLC中,p63和p73蛋白表达之间呈显著正相关(P=0.000 1),p73与p53蛋白表达之间无显著相关性(P＞0.05).结论p53基因家族可能与NSCLC的发生发展有关.p63和p73蛋白有不同于p53蛋白的生物学功能,二者可能均起癌基因的作用.     

胃癌中hMSH2、p53和PCNA表达的相关性及意义

目的:探讨胃癌中hMSH2、p53和PCNA表达的相关性及意义.方法:采用免疫组织化学SP法,检测胃癌、癌旁和胃炎粘膜中hMSH2、p53和PCNA表达情况.结果:1)3种基因产物在胃癌中的阳性率均显著高于非癌组织,其中,p53和PCNA在低分化癌中的阳性率显著高于高分化癌,有淋巴结转移者显著高于无转移者(P＜0.05).2)胃癌中hMSH2/PCNA及p53/PCNA表达均呈正相关(P＜0.05).结论:hMSH2、p53和.PCNA的异常表达及hMSH2与PCNA之间的相互调节可能与胃癌的发生发展密切相关.     

p53蛋白与增殖细胞核抗原在胃癌中表达及其临床意义

目的研究p53、增殖细胞核抗原(PCNA)在胄癌中的表达及其与临床病理和预后的关系.方法采用免疫组化技术(S-P法)对33例胃癌组织及20例正常胃粘膜组织进行p53、PCNA检测.结果 p53、PCNA阳性表达率分别为57.6%,93.9%;PCNA强阳性表达率为67.7%;p53阳性表达率及PCNA强阳性表达率均与胃癌组织类型、浸润深度、淋巴结转移密切相关.差异具有显著性(P＜0.05);p53蛋白表达与PCNA强阳性表达呈正相关.结论联合检测p53和PCNA对判断胃癌的恶性程度、浸润深度和淋巴结转移趋势和预后具有重要价值.     

p63和TTF-1在肺癌组织中的表达及鉴别诊断意义

目的:探讨p63和TTF-1蛋白在肺癌不同组织类型中的表达特点及鉴别诊断的价值。方法:应用免疫组织化学S-P法检测116例原发性肺癌组织中p63和TTF-1蛋白的表达情况。结果:p63在肺鳞癌中阳性表达率为100%(47/47),而在其他组织类型肺癌中基本不表达。TTF-1在肺小细胞癌和肺腺癌中阳性表达率分别为93.3%(14/15)和93.5%(43/46),而在鳞癌中未见表达。结论:p63可作为肺鳞状上皮源性肿瘤标记物,是判断鳞癌的可靠指标。TTF-1可作为肺小细胞癌、腺癌的特异性标记。联合检测p63和TTF-1对肺癌不同组织类型的诊断、鉴别诊断具有较高的实用价值,尤其可作为鉴别分化差的鳞癌、腺癌和小细胞癌的指标。     

P21WAF1/CLP1在甲状腺癌组织的表达及临床意义

[目的]探讨p21WAF/CLP与p53基因在甲状腺癌中的表达与病理类型和分化程度的关系。[方法］应用免疫组化技术检测甲状腺癌组织中的抑癌基因p2lWAF/CLPl及p53基因的表达。统计学采用x2检验。[结果]甲状腺腺瘤(TT),甲状腺乳头状癌(PTC)、滤泡状癌(FTC)、未分化癌(UDC)组织p21蛋白阳性率分别为66.7％、62 5％、58.3％、50.0％,各组之间无显著差别(P＞0.05);高分化癌(WDC)、低分化癌(PDC)、UDC组织p21蛋白阳性率分别为67.7％、46.2％、50.0％,各组之间无显著差别(P＞0.05);PTC、FTC、UDC组织p53蛋白阳性率分别为31.3％、25.0％、64.3％,UDC与PTC、FTC之间存在显著差别(P＜0 05);WDC、PDC、UDC组织p53蛋白阳性率分别为l6.1％、61.5％、64.3％,WDC的阳性率明显低于PDC、UDC(P＜0.05);p53蛋白阳性的癌组织p21蛋白阳性率为40.9％(9/22),p53蛋白阴性的癌组织p21蛋白阳性率为69.4％(25/36),两者之间存在显者差异(P＜0 05)。[结论］甲状腺癌中p21WAF/CLP的表达与其病理类型和分化程度无关,而p53基因的表达与甲状腺癌病理类型和分化程度有关,p2WAFl/CLPl作为一新的抑癌基因,其表达相当程度上依赖于p53蛋白。     

p21^waf1、p53和PCNA在肺癌组织中的表达

目的了解肺癌组织中p21waf1、p53和PCNA蛋白表达情况,研究肺组织良恶性增殖的不同机制.方法运用免疫组织化学方法,检测76例肺癌组织p21waf1、p53及PCNA的表达情况,分析三种蛋白表达与肺癌临床特征的关系,并与59例肺良性疾病组织相关指标进行比较.结果(1)肺癌组织p21waf1和p53蛋白表达阳性率分别为75%和47.37%,PCNALI值为0.44±0.32,均显著高于肺良性疾病组织(分别为35.59%、3.39%和0.09±0.14).(2)肺癌组织p21waf1、p53和PCNA蛋白表达与肺癌临床分期及细胞分化程度无关.各型肺癌之间p21waf1、p53蛋白表达也无明显差异.(3)肺鳞癌PCNALI值为0.51±0.32,明显高于小细胞肺癌(0.30±0.36).(4)肺癌组织p21waf1和p53蛋白表达与PCNA无关,p21waf1和p53蛋白表达之间也未发现相关性.而肺良性疾病组织p21waf1和p53蛋白表达与PCNA相关.结论(1)肺癌组织p21waf1与p53蛋白表达明显上调.(2)肺癌细胞增殖程度很高.(3)肺鳞癌细胞DNA损伤修复能力可能高于小细胞肺癌.(4)肺良性疾病细胞增殖过程中,p21waf1、p53及PCNA三种蛋白有协调作用,而在肺癌细胞则否.     

p53基因突变产物在肺癌组织和胸水的表达及其意义

应用免疫组化方法研究了p53蛋白在肺癌组织和胸水细胞中的表达情况。发现p53蛋白在常规石蜡切片和胸水中保存较好,在肺癌组织中表达的总阳性率为66.3%,正常细胞中未见p53蛋白表达;肺癌有淋巴结转移组p53蛋白的阳性率(75.5%)高于无淋巴结转移组(55.8%)(P＜0.05);Ⅲ—Ⅳ期肺癌p53蛋白的阳性率(82.9%)高于Ⅰ—Ⅱ期肺癌(56.1%)(P＜0.01)。结果表明,p53基因突变与肺癌的发生、淋巴结转移及进展有关,突变型p53蛋白是肺癌较特异的肿瘤标记物。     

肺癌MTS1/p16和p53表达与细胞增殖

（目的〕研究肺癌中MTS1／p16和p53基因产物的表达与细胞增殖的关系。〔方法〕应用SP免疫组织化学方法研究62例肺癌组织中p16蛋白和p53蛋白的表达情况，并进行增殖细胞核抗原检测，计算细胞增殖指数（PI）。(结果)62例肺癌组织中p16蛋白和p53蛋白阳性率分别为58．1％和59、7％。晚癌p16蛋白的阳性率明显高于小细胞癌（p＜0.05）；淋巴结转移阳性组p16蛋白的表达显著低于阴性组（P＜0．05）；PI分级为巨级的p16蛋白表达显著高于Ⅳ级（p＜0.05）。不同组织类型肺癌中p53蛋白的表达未见明显差异，淋巴结转移阳性组p53蛋白的表达高于阴性组（p＜0.01〕；不同PI分级中p53蛋白的表达，N级明显高于1级和Ⅱ级（P＜0．05），三级明显高于1级（p＜0．05）和Ⅱ级（P＜0.01）。p16蛋白低表达和p53蛋白过度表达之间未见明显相关性。（结论〕提示p16蛋白低表达和p53蛋白过度表达均有促进肺癌细胞增殖的作用，p16蛋白的表达与肺癌的细胞分化有关，p53蛋白过度表达对肺癌细胞的转移起重要作用。抑癌基因p53对MTS1／p16基因无明显调控作用。检测p53蛋白表达可作为肺癌诊断的一项新指标。     

肺癌端粒酶活性、p53、PCNA蛋白表达的联合检测及其临床意义

目的 探讨端粒酶活性p53基因和增殖细胞核抗原（PCNA）在肺癌发生发展中的作用，以及端粒酶与临床病理特点及p53,PCNA蛋白之间的关系。方法 TRAP法检测59例肺癌和35例癌旁正常组织中端粒酶活性，采用免疫组织化学法检测44例肺癌组织和35例癌旁正常组织中p53和增殖细胞核抗原（PCNA）蛋白的表达，并对端粒酶活性与临床病理特征以及p53PCNA蛋白表达的关系进行分析。结果 肺癌组织端粒酶活性,p53和PCNA蛋白阳性率分别为81．3％（48／59）、77．2％（34／44）、79．5％（35／44）；癌旁正常组织端粒酶活性,p53和PCNA蛋白阳性率分别为20．0％（7／35）、8．6％（3／35）、14．3（5／35），肺癌组织显著高于癌旁正常组织，P＜0．01，肺癌组织不同的病理类型之间、TNM各期之间以及不同的细胞分化程度之间端粒酶活性的阳性率未发现明显的差异（P＞0．05）；端粒酶活性与p53,PCNA蛋白表达之间无明显相关性。结论 端粒酶活化参与各型、各期和不同分化程度肺癌的发生发展；端粒酶在肺癌组织高表达和正常肺组织低表达的特性，有望使端粒酶成为肺癌诊断的1种标志物；p53和PCNA均参与肺癌的发生发展过程；端粒酶与p53和PCNA无明显相关性，提示肺癌是由多基因参与的疾病。     

p29ING4 and p28ING5 bind to p53 and p300, and enhance p53 activity

We identified and characterized two new ING family genes, p29ING4 and p28ING5,coding for two proteins of 249 and 240 amino acids, respectively. Both p29ING4 and p28ING5 proteins have a plant homeodomain finger motif also found in other ING proteins, and which is common in proteins involved in chromatin remodeling. p29ING4 or p28ING5 overexpression resulted in a diminished colony-forming efficiency, a decreased cell population in S phase, and the induction of apoptosis in a p53-dependent manner. Both p29ING4 and p28ING5 activate the p21/waf1 promoter, and induce p21/WAF1 expression. p29ING4 and p28ING5 enhance p53 acetylation at Lys-382 residues, and physically interact with p300, a member of histone acetyl transferase complexes, and p53 in vivo. These results indicate that p29ING4 and p28ING5 may be significant modulators of p53 function.     

INK4系列抑癌基因纯合子缺失、甲基化与白血病预后的实验研究

 目的　研究INK4系列抑癌基因纯合子缺失、甲基化与白血病预后的关系。方法　采用聚合酶链反应（PCR）研究p16基因家族在白血病中纯合子缺失,应用甲基化敏感限制内切酶HpaⅡ结合PCR技术研究白血病患者p16、p15、p18、p19 基因甲基化状况,用单因素、多因素Logistic回归分析其基因失活与急性白血病（AL）预后的关系。结果　基因表达组治疗有效27例（84.38 %）,基因失活组治疗有效11例（28.95 %）,基因表达组治疗有效率明显高于基因失活组（P＜0.001）。单因素、多因素Logistic回归分析结果显示p16、p15 基因失活化疗有效率明显低于基因表达组。结论　p16、p15基因失活可作为AL病程进展、复发、预后的指标之一。     

p53基因家族的新成员 p73和 p63

p53作为广泛存在的肿瘤抑制基因,最近发现了其家族成员：p73和p63。它们在结构和功能上具有相似性,均能触发细胞周期停滞,诱导凋亡,但它们在肿瘤抑制和组织发育中起着截然不同的作用,深入了解它们彼此之间的相似性和差异将对理解肿瘤发生的机制产生重要的影响。本文总结了最近几年来此领域内的最新进展。另外,p73和p63在C端的SAM结构说明它们可能参与组织的发育过程。     

p53 protein variants: structural and functional similarities with p63 and p73 isoforms

Since its discovery in 1979, many studies have reported that the p53 tumour suppressor protein could be expressed in the form of products smaller than those predicted by the full-length amino-acid sequence. These products differ from full-length p53 in their N- or C-terminal regions, but generally conserve the central, DNA-binding domain. They appear to be expressed at rather low levels and to be restricted to particular cell types and/or physiological circumstances, suggesting that they play very narrow and specific roles. Several mechanisms have been proposed to explain their timely occurrence, including alternative splicing, internal initiation of translation or proteolytic cleavage. A precise assessment of the various 'p53 isoforms' reveals striking similarities with several isoforms of the p53 homologous proteins p63 or p73, suggesting that regulated production of specific, N- or C-terminal variants may be a 'trademark' of all family members. In this review, we summarize the published evidence on the structure, mode of production, expression and function of the p53 isoforms, and discuss their properties in the light of recent data on the structure and function of p63/p73 isoforms.     

Tumorigenesis in p27/p53- and p18/p53-double null mice: functional collaboration between the pRb and p53 pathways

Mice lacking both p18(Ink4c) and p27(Kip1) develop a tumor spectrum similar to pRb(+/-) mice, and loss of p53 function accelerates tumorigenesis in pRb(+/-) mice. We hypothesized that codeletion of either p18 or p27 in conjunction with p53 deletion will also accelerate tumorigenesis. Mice lacking both p18 and p53 develop several tumors not reported in either single null genotype, including hepatocellular carcinoma, testicular choriocarcinoma, hemangiosarcoma, leiomyosarcoma, fibrosarcoma, and osteosarcoma. Mice lacking both p27 and p53 exhibit a decreased lifespan and develop unique tumors, including papillary carcinoma of the colon, hemangiosarcoma, and leiomyosarcoma. In both p18/p53 and p27/p53 double null genotypes, the incidence and spectra of tissues that develop lymphoma are also increased, as compared to the single null genotypes. The development of p27/p53 double null colon tumors correlates with secondary changes in cell-cycle protein expression and CDK (cyclin-dependent kinase) activity, perhaps contributing to the progression of colorectal cancer. We concluded that p18 and p27 can, not only functionally collaborate with one another, but also can independently collaborate with p53 to modulate the cell cycle and suppress tumorigenesis in a tissue-specific manner.     

Role of p53 family members p73 and p63 in human hematological malignancies

Abstract p53, mutated in over half of human cancers and about 13% of all hematological malignancies, maintains genomic integrity and triggers cellular senescence and apoptosis of damaged cells. In contrast to p53, the homologs p73 and p63 play critical roles in development of the central nervous system and skin/limbs, respectively. Moreover, dependent on the context they can exert tumor suppressor activities that cooperate with p53. Unlike p53, p73 and p63 are rarely mutated in cancers. Instead, up-regulation of the anti-apoptotic dominant-negative ΔNp73 and ΔNp63 isoforms is the most frequent abnormality in solid cancers. In hematological malignancies the most frequent p73 defect is promoter methylation and loss of expression, associated with unfavorable clinical outcomes. This suggests an essential tumor suppressor role of p73 in blood cells, also supported by genetic mouse models. Many therapeutic approaches aiming to restore p73 activity are currently being investigated. In contrast, the most frequent p63 abnormality is protein overexpression, associated with higher disease grade and poorer prognosis. Surprisingly, although available data are still scarce, the emerging picture is up-regulation of transactivation-competent TAp63 isoforms, suggesting a tumor-promoting role in this context.     

p73和p63蛋白在胰腺癌组织中过表达的意义

目的:探讨p53家族新成员p73和p63蛋白在胰腺癌组织中过表达的意义。方法:应用免疫组化LSAB法检测75例人胰腺癌组织中p73和p63蛋白的过表达。结果:p73和p63蛋白在人胰腺癌中的过表达率分别为46·7%(35/75)和42·7%(32/75),p73蛋白在胰腺囊腺癌中的过表达率88·9%(8/9)明显高于导管腺癌41·8%(23/55),P=0·009,且p73蛋白过表达与胰腺癌淋巴结转移、肿瘤大小、神经侵犯和p53表达呈显著负相关性,P<0·05;在腺鳞癌或腺癌伴鳞状上皮化生中p63蛋白的过表达率(100%,13/13)明显高于导管腺癌(40·0%,22/55),P=0·007,但p63蛋白过表达与胰腺癌临床病理学指标及p53和增殖细胞核抗原(pro-liferatingcellnuclearantigen,PCNA)表达间无明显相关性。结论:p73蛋白低表达可能在胰腺癌发生中起重要作用;p63蛋白过表达与腺鳞癌或腺癌鳞化有关。     

A Comparative Study of Glioma Cell Lines for p16, p15, p53 and p21 Gene Alterations

A total of 10 glioma cell lines were examined for alterations of the p16, p15, p53 and p21 genes, which are tumor suppressor genes or candidates with direct or indirect CDK-inhibitory functions. Genetic alterations (deletions or mutations) were frequently seen in the p16, p15 and p53 genes in these cell lines, but not in the p21 gene. When the states of the p16, p15 and p53 genes were compared among cell lines, all the cell lines showed abnormalities in at least 1 gene, often in 2 or 3 genes coincidentally, suggesting that dysfunction of these genes is closely related to glioma cell growth. Although alteration of all 3 genes was most frequent, there were cell lines having either p16/p15 or p53 or p16 and p53 gene alterations, suggesting that the time order of these genetic alterations was variable depending on the cell line. Among cell lines examined, one with homozygous p53 gene deletion seemed of particular practical value, since such a cell line might be useful in various studies, including investigation of the functions of various mutant p53 genes in the absence of heteromeric protein formation. On examination of the primary tumor tissues, the same alterations of the p16/p15 and p53 genes as detected in the cell lines were demonstrated in all 6 cases examined: p16/p15 gene deletion in 1, p16 gene mutation in 1 and p53 gene mutations in 5 cases. This suggested that the p16/p15 and the p53 gene alterations and their combinations in at least some glioma cell lines reflected those in the primary glioma tissues.