原发性输尿管癌p53、c-erbB-2和nm23表达与临床病理特征的关系

目的　研究输尿管癌 p5 3、c- erb B- 2、nm2 3基因的不同表达与其临床病理因素的关系。方法　应用免疫组化 S- P法检测 p5 3、c- erb B- 2、nm2 3等基因蛋白在 2 4例原发性输尿管癌中的表达情况 ,并对其结果与临床病理各参数的关系进行分析。结果　正常输尿管上皮三种基因蛋白均呈阴性 ,在输尿管癌中 p5 3、c- erb B- 2、nm2 3的阳性表达率分别为 41.6 % (10 /2 4)、5 4.1% (13/2 4)和 6 6 .7% (16 /2 4)。 p5 3、c- erb B- 2高表达与输尿管癌分化程度低、浸润至浆膜或浆膜外、生存期短等几项临床病理特征相关。nm2 3的表达与输尿管癌病理分级、浸润深度、临床预后无密切关联。结论　 p5 3、c- erb B- 2和 nm2 3基因参与了输尿管癌的发生发展过程 ,p5 3、c- erb B- 2蛋白表达可以作为输尿管癌生物学行为和预后判断的参考指标 ,两指标同时检测预示作用更可靠     

骨肉瘤组织中Vimentin,Actin,PCNA及相关癌基因的表达

目的 :分析 Vimentin、Actin、PCNA、p5 3基因、C- erb B- 2及 nm 2 3基因在骨肉瘤中的表达及其相关性。方法 :应用免疫组化技术分别检测 Vimentin、Actin、PCNA、p5 3、C- erb B- 2及 nm 2 3基因蛋白在 33例骨肉瘤中的表达 ,其中 19例获得平均 1a 7个月的随访。结果 :Vim entin阳性表达率为 87.9% ,Actin为 5 4.5 % ,PCNA为6 6 .7% ,p5 3为 5 7.6 %、C- erb B- 2为 15 .2 % ,nm 2 3增强表达率为 90 .9%。经过分析 ,除 PCNA和 p5 3表达呈平行关系外 ,余各项指标间无相关性 ;Vimentin、Actin阳性表达与预后无关 ;p5 3、PCNA和 nm 2 3基因蛋白表达与预后呈正相关。结论 :Vim entin、Actin可用于研究骨肿瘤的组织起源 ;PCNA和 p5 3在骨肉瘤中表达呈平行关系 ,与预后呈正相关 ;nm2 3基因在骨肉瘤中无转移抑制基因的功能。     

宫颈鳞癌组织HPV16/18 感染与多个癌基因产物表达的研究

 目的　探讨 HPV1 6、1 8的感染及多个癌基因的激活在宫颈鳞癌发生发展中的作用。方法　采用免疫组化方法对 1 9例慢性宫颈炎、40例宫颈上皮内瘤变 ( CIN)及 70例浸润性宫颈鳞癌进行 E6、PCNA、p5 3、p2 1 ras和 c- myc蛋白检测。结果　CIN 和浸润性宫颈鳞癌组 E6、PCNA、p5 3、p2 1 ras阳性率明显高于慢性宫颈炎、CIN 和 CIN ,而其 c- myc阳性率则明显低于其它几组。结论　 HPV1 6、1 8的感染及 PCNA、p5 3、p2 1 ras、c- myc的异常表达与宫颈鳞癌的发生发展密切相关。     

p53蛋白在非小细胞肺癌中的表达及意义

目的 :探讨非小细胞肺癌中p5 3蛋白的表达及意义。方法 :应用SP免疫组化法检测 5 2例肺癌组织中p5 3蛋白的表达。结果 :非小细胞肺癌中p5 3蛋白阳性表达率为 78 8% ,其中鳞癌与腺癌分别为 75 0 %和 82 1% ,p5 3蛋白阳性病人的预后较阴性者差。结论 :p5 3蛋白在非小细胞肺癌中有较高的表达 ,有可能作为评估预后、指导治疗的指标。     

非小细胞肺癌分子病理学的研究

目的　探讨上海地区人肺癌发生及发展过程中的分子病理学模式。方法　采用DNAslotblot、PCR、PCR SSCP等方法 ,先后检测 2 0 0例NSCLC组织DNA标本中C erbB2、C myc、EGFR癌基因扩增 ,抑癌基因中p5 3基因外显子 5～ 8点突变、p15基因的纯合性丢失 ,以及某些与肺癌相关的染色体 3p及 17p13 .3位点的杂合性丢失 (LOH)。结果　多种癌基因共扩增率与肺癌TNM分期呈正相关 (P <0 .0 0 1)。p5 3基因外显子 5～8点突变率为 49.2 % ( 3 1/63 ) ,其中以外显子 8为主。肺癌组织中p15基因出现高频率纯合性丢失 ,并与肺癌TNM分期密切相关 (P <0 .0 0 5 )。 17p13 .3杂合性丢失率为 40 % ( 8/2 0 ) ,并与p5 3基因的点突变相关联。 3p14及 3p2 5位点处的杂合性总丢失率可高达 66.7% ( 10 8/162 ) ,其中肺腺癌中 3p14及 3p2 5二位点的共丢失率明显高于鳞癌 (P <0 .0 5 )。结论　根据上述结果 ,提出上海市区居民的非小细胞肺癌分子病理学的初步模式 :癌前期病变时已可出现 3p丢失 ,原位癌时以p5 3基因突变及 17p13 .3丢失为主 ,浸润癌及向转移性癌过渡时有多种癌基因C myc、C erbB2及EGFR癌基因共扩增 ,并出现p15基因纯合性丢失。     

非小细胞肺癌P63基因的表达与3q27-q29变化的关系研究

目的　研究p63基因在非小细胞肺癌中的蛋白表达水平及与 3号染色体长臂 3 q2 7 3 q2 9区域改变的关系。方法　采用组织芯片技术构建 12 2例原发性非小细胞肺癌石蜡包埋标本组织芯片 ,并应用免疫组织化学方法检测 p63基因蛋白表达情况。应用比较基因组杂交 (CGH)技术分析 70例原发性肺鳞癌和肺腺癌标本染色体的改变。同时分析p63基因的蛋白表达与 3号染色体长臂末端改变的关系。结果　 12 2例非小细胞肺癌 p63免疫组化染色结果显示 :5 9例鳞癌中 5 1例 ( 86.44 % ) p63免疫组化染色为阳性 ;3例大细胞肺癌中 2例 ( 66.66% )为阳性 ;60例腺癌仅有 1例 ( 1.67% )为阳性。p63蛋白的阳性表达率与患者的年龄、性别、肿瘤的分级、肿瘤的转移以及预后生存率无关 (P >0 .0 5 )。 70例非小细胞肺癌CGH分析结果发现有3 2例 3 q2 7 q2 9区域出现DNA扩增 ,3 0例鳞癌中 2 4例出现 3 q2 7 q2 9区域DNA拷贝数目的增加 ,40例腺癌仅 8例发现 3q2 7 q2 9区域DNA获得。p63免疫组化阳性率与 3q2 7 q2 9区域的改变比较分析结果显示 :2 4例鳞癌 p63阳性者中 ,2 3例 ( 95 .83 % ) 3 q2 7 q2 9区域有显著的获得 ,1例 p63基因蛋白表达呈阳性的腺癌患者的 3 q2 7 q2 9区域有DNA拷贝数目的增加。 结论　研究结果提示 p63免疫组化阳性     

CD44v3在非小细胞肺癌中的表达以及对生存率的预测价值

目的　研究CD44v3在非小细胞肺癌中的表达及其对患者生存的预测价值。方法　采用RT PCR和免疫组化SP法等技术 ,对 5 2例非小细胞肺癌患者手术标本进行CD44v3检测 ,并随访。结果　两种方法测得 5 2例肺癌CD44v3阳性表达率分别为 5 7.7% (SP法 )和 71.2 % (RT PCR法 )。其中鳞癌CD44v3的阳性表达率显著高于腺癌 (P <0 .0 1) ,低分化者显著高于高分化和中分化 (P <0 .0 5 ) ,有淋巴结转移的肺癌患者显著高于无淋巴结转移者 (P <0 .0 5 ) ,Ⅲ期患者显著高于Ⅰ期和Ⅱ期 (P <0 .0 5 )。多元Logistic回归分析显示 ,CD44v3阳性表达率和 p TNM分期对生存率有显著影响 (P <0 .0 5 )。 结论　CD44v3普遍存在于非小细胞肺癌组织中。检测CD44v3有助于预测肺癌患者术后生存。     

癌基因K-ras、C-myc、erbB-2的激活与非小细胞肺癌组织学类型相关性研究

为探讨癌基因突变、扩增与非小细胞肺癌发生及类型间的关系，应用核酸杂交的方法检测了63伤1肺手术标本中（腺癌19例，鳞癌23例，腺鳞癌3例，大细胞癌3例，小细胞癌3例，其它12例）三种癌基因（c－myc、K－ras、erbB－2）的扩增及突变。实验结果显示：50％（24／48）的非小细胞癌分别出现三种癌基因的激活；31．6％（6／19）的腺癌出现K－ras12密码子突变及K－ras扩增；30．4％（7／23）的鳞癌有erbB－2扩增；10．4％（5／48）的非小细胞癌存在c－myc扩增。提示K－ras突变可能为肺腺癌所特有，而erbB－2扩增是肺鳞癌发生的重要因素。     

C-erbB-2、p16、nm23-H_1蛋白在鼻咽癌中表达的相关性及与预后的关系

目的 :探讨癌基因 C- erb B- 2、抑癌基因 p16和转移抑制基因在鼻咽癌中表达的相关性及与鼻咽癌发生、发展和预后的关系。方法 :采用 SPTM(过氧化酶标记的链霉卵白素 )免疫组化法 ,对 12 5例鼻咽癌组织中三种基因蛋白进行检测。结果 :1C- erb B- 2 ,p16 ,nm 2 3- H1 在低分化鳞癌中的阳性率分别为 5 8.75 %、45 %、32 .5 % ,在未分化癌中阳性表达率为 86 .6 7%、31.11%、6 0 % ,组间差异有显著性 (P<0 .0 1) ;2三种基因均在不同临床分期鼻咽癌组织中表达差异有显著性 (P<0 .0 1) ;3 p16蛋白表达与有、无淋巴结转移显著性差异 ;4同时存在 C- erb B-2蛋白阴性 ,p16 ,nm 2 3- H1 阳性者共 30例 ,其中 5年存活率为 86 .6 7% (2 6 / 30 ) ,然而 ,C- erb B- 2 ( ) ,p16 ,nm 2 3- H1均 (- )者共 48例 ,5年存活率仅 31.2 5 % (16 / 48) ,差异有显著性。结论 :三种基因均与鼻咽癌的发生、发展及预后密切相关 ,癌基因的过度表达和抑癌基因及转移抑制基因的表达缺失的负相关性 ,可作为判断预后的指标之一。     

p16、p53、p21ras和C-erbB-2在非小细胞肺癌中的表达与病理学相关性研究

 目的　研究p16、p53、p21ras和C-erbB-2在非小细胞肺癌中的表达与临床病理学关系。方法　用免疫组化方法检测87例肺癌中p16、p53、p21ras和C-erbB-2的表达,和组织学分型、分化程度、淋巴结转移、肿瘤大小及TNM分期的关系。结果　p16、p53表达阳性与淋巴结转移及临床分期等因素有关。p21ras、C-erbB-2均在腺癌表达阳性并与分化程度相关。结论　p16表达阳性与肺癌的发生与发展有关,与肿瘤分化较好、恶性程度较低等预后好的生物学因素有关。p21ras表达与组织学分型及分化程度有关。p53表达阳性表明肿瘤分化差及淋巴结转移的高危性。C-erbB-2蛋白可作为腺癌的一种标志物,且提示预后较差。     

INK4系列抑癌基因在白血病中纯合子缺失的实验研究

目的：探讨p16基因家族失活与白血病发生、发展及预后的关系。方法：PCR检测p16、p15、p18、p19基因在白血病中纯合子缺失。结果：p16、p15基因外显子1在AL组纯合子缺失率分别为22．37％、17．05％,在ALL组为45．95％、32．43％,在ANLL组均为5．88％;在CML的慢性期均为0。p16、p15基因外显子2在AL组纯合子缺失率分别为12．5％、5．68％;在ALL组为24．32％、10．81％;在ANLL组为3．92％、1．96％;在CML和对照组二者均无缺失。p18、p19基因外显子1在AL组纯合子缺失率分别为1．14％、0;在ALL组分别为2．70％、0;在ANLL和对照组均无缺失。结论：p16、p15基因纯合子缺失在AL的发生频率较高,ALL缺失率明显高于ANLL,复发一LLL组基因失活率最高。p16、p15基因纯合子缺失是AL,尤其是ALL的发病重要因素之一。p18、p19基因在AL组中几乎未见纯合子缺失。在ALL中,p16纯合子缺失率高于p15纯合子缺失率。两个基因的纯合子缺失常伴随存在。在ANLL中,p16、p15基因的纯合子缺失均少见。     

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.     

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。     

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。     

NBP is the p53 homolog p63

We previously identified a non-p53, p53-responsive DNA element (p53RE)-binding protein named NBP, functionally analogous to p53, from human cervical carcinoma Hela cells. Here we report a biochemical study demonstrating that this activity is the recently cloned p53 analog p63. NBP was purified through conventional and DNA affinity chromatography to apparent homogeneity with a prominent polypeptide migrating in between the 43 and 68 kDa positions on a SDS gel. This polypeptide immunoreacted with monoclonal anti-p63 but not anti-p53 or anti-p73 antibodies. Also, NBP co-purified with p63 through each step of fractionation, as detected with anti-p63 antibodies. DNA-protein complexes formed with purified NBP and p53RE-containing oligomers derived from the p21(waf1) promoter were supershifted by anti-p63 but not anti-p53 antibodies. Thus, these results demonstrate that NBP is encoded by the p53 homolog p63 gene.     

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.     

原发性肝细胞癌中p21WAF1及p16INK4a 的表达与HBV的关系

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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.     

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.