ras、p53基因突变和蛋白表达与尿路上皮肿瘤预后的关系

目的　研究ｒａｓ、ｐ５３ 基因突变和蛋白表达及ＤＮＡ 倍体异常与尿路上皮肿瘤预后关系。　方法　应用ＰＣＲＳＳＣＰ和流式免疫荧光法检测５２ 例尿路上皮肿瘤Ｋｒａｓ 和ｐ５３ 基因突变及蛋白表达和ＤＮＡ 含量变化。　结果　Ｋｒａｓ、ｐ５３ 基因突变分别为５ 例（９ ．６ ％ ） 和２２ 例（４２ ．３ ％ ） ;ｒａｓ ｐ２１ 、ｐ５３ 蛋白阳性表达分别为３６ 例（６９ ．２ ％ ） 和３３ 例（６３ ．５ ％ ） ;异倍体和二倍体肿瘤分别为２２ 例（４２ ．３ ％ ） 和３０ 例（５７ ．７ ％ ） ;ＤＮＡ 倍体异常、ｐ５３ 基因突变及ｒａｓ ｐ２１ 和ｐ５３ 蛋白表达与肿瘤病理分级、分期及预后密切相关（ Ｐ＜ ０ ．０１） ;异倍体肿瘤其ｐ５３ 基因突变、ｒａｓ ｐ２１ 和ｐ５３ 蛋白表达率均明显高于二倍体肿瘤（ Ｐ＜ ０ ．０１） 。　结论　ＤＮＡ 倍体异常、ｐ５３ 基因突变及ｒａｓ ｐ２１ 和ｐ５３ 蛋白表达对尿路上皮肿瘤预后判断有重要意义。     

甲状腺癌中p53基因248位密码子突变及其蛋白过度表达的临…

对５４例甲状腺癌标本分别用ＤＮＡ聚合酶链反应－限制性片段长度多态性分析（ＰＣＲ－ＲＦＬＰ）法检测ｐ５３基因第２４８位密码子的突变及免疫组化法检测ｐ５３蛋白，并用时序检验分析ｐ５３基因突变和ｐ５３蛋白表达与甲状腺癌的关系。结果发现，甲状腺癌中１５例（２７．８％）有ｐ５３基因突变和３０例（５５．６％）有ｐ５３蛋白过度表达，临床分期晚、分化程度低的甲状腺癌突变率较高，ｐ５３抑癌基因突变和ｐ５３蛋白阳性病     

大肠癌患者粪便中C-erbB-2扩增和p53突变的检测及临床意义

目的　建立大肠癌患者粪便 Ｃｅｒｂ Ｂ２ 扩增及ｐ５３ 突变的检测方法,探讨大肠癌的基因诊断的临床意义。 方法　以差异聚合酶链反应（ Ｐ Ｃ Ｒ） 、 Ｐ Ｃ Ｒ单链构象多态性分析（ Ｓ Ｓ Ｃ Ｐ） 银染技术,分别检测大肠癌患者粪便 Ｃｅｒｂ Ｂ２ 扩增及ｐ５３ 突变。 结果　１４ 例中查出 Ｃｅｒｂ Ｂ２ 扩增７ 例（５０ ％ ） ,ｐ５３ 突变８ 例（５７ ％ ） ,二基因联合检出１１ 例（７９ ％ ） 。２ 例粪便潜血试验（ Ｆ Ｏ Ｂ Ｔ） 阴性中,１ 例 Ｃｅｒｂ Ｂ２扩增及ｐ５３ 突变均阳性,另１ 例ｐ５３ 突变阳性。 结论　联合多基因检测能对大肠癌的基因诊断提供更多的帮助,粪便 Ｄ Ｎ Ａ 标本的 Ｃｅｒｂ Ｂ２ 扩增及ｐ５３ 突变分析可为筛查尚无出血或 Ｆ Ｏ Ｂ Ｔ 假阴性的大肠癌及大肠癌高危个体的一种新的有效手段。     

肾癌p53蛋白表达及其点突变相关研究

目的探讨肾细胞癌发病机制中致癌基因突变规律。方法以ｐ５３蛋白表达及其点突变率为实验观测指标,对以往１１６例各类肾肿瘤以免疫组化ＡＢＣ法检测病理切片的ｐ５３表达。并以ＰＣＲＳＳＣＰ观测５９外显子点突变,并以１２例非瘤新鲜肾组织作对照。结果三类恶性肾细胞癌的ｐ５３阳性表达率介于４２％～５４％之间,且随癌细胞恶性度增加及分期增加而升高,而良性瘤及正常肾组织为阴性。点突变率介于２５％～７１％之间,与恶性度及分期相关。第９外显子点突变率明显高于其它区域,良性瘤及正常肾无点突变。结论肾癌发病机制与ｐ５３基因突变密切相关。ｐ５３蛋白阳性表达与其点突变相辅并存,协同一致,二者阳性率与生存期及预后密切相关     

甲状腺癌中p53基因248位密码子突变及其蛋白过度表达的临床意义

对５４例甲状腺癌标本分别用ＤＮＡ聚合酶链反应－限制性片段长度多态性分析（ＰＣＲ－ＲＦＬＰ）法检测ｐ５３基因第２４８位密码子的突变及免疫组化法检测ｐ５３蛋白，并用时序检验分析ｐ５３基因突变和ｐ５３蛋白表达与甲状腺癌的关系。结果发现，甲状腺癌中１５例（２７８％）有ｐ５３基因突变和３０例（５５６％）有ｐ５３蛋白过度表达，临床分期晚、分化程度低的甲状腺癌突变率较高，ｐ５３抑癌基因突变和ｐ５３蛋白阳性的病例有较高的复发率和死亡率。表明ｐ５３基因突变及其蛋白过度表达在甲状腺癌的发生和发展过程中起重要作用，是预后不良的标志。     

应用PCR－SSCP研究胆道肿瘤P53基因的突变

应用ＰＣＲ－ＳＳＣＰ（ＤＮＡ聚合酶链式反应－单链构象多态性）方法检测了１２例胆道肿瘤（胆管癌７例，胆囊癌３例，壶腹癌２例）的Ｐ５３基因第５、７外显子，结果发现２例胆管癌和１例胆囊癌Ｐ５３基因有突变。采用免疫组化方法对２２例胆管癌和８例癌旁组织Ｐ５３蛋白产物进行了检测，染色阳性者分别为１１例（５０％）和０。结果提示胆道肿瘤的发生与Ｐ５３基因的突变有关。     

人乳头瘤病毒DNA相关序列与p53基因突变对大肠癌生物学行为的影响50例报告

目的探讨人乳头瘤病毒（ＨＰＶ）ＤＮＡ相关序列与ｐ５３基因突变及ｐ５３蛋白表达的关系及其对大肠癌生物学行为的影响。方法采用ＰＣＲ方法检测大肠癌及癌旁组织、肝转移灶中ＨＰＶＤＮＡ相关序列。并应用ＰＣＲＳＳＣＰ及免疫组化技术分别检测ｐ５３基因突变及ｐ５３蛋白表达。结果在５０例大肠癌中,检出ＨＰＶ１６、１８ＤＮＡ相关序列２６例（５２０％）,其中ＨＰＶ１６ＤＮＡ４例（８０％）,ＨＰＶ１８ＤＮＡ２２例（４４０％）。ｐ５３基因突变率为５６０％。ｐ５３蛋白表达阳性率为４２０％。ＨＰＶＤＮＡ相关序列与ｐ５３基因突变及ｐ５３蛋白表达呈正比关系。结论ＨＰＶＤＮＡ相关序列可促进细胞转化、致ｐ５３基因突变、抑制细胞的凋亡,与大肠癌的发生发展有密切关系。     

胆汁中突变基因与大肠癌肝转移关系的研究

目的 探讨胆汁中的点突变基因对诊断大肠癌肝转移的价值。方法 采用ＰＣＲ－ＳＳＣＰ及ＤＮＡ测序技术,检测大肠癌患者原发灶、肝转移灶、胆汁中相同突变位点的ｐ５３、ｋ－ｒａｓ基因。结果 在５０例大肠癌中,检出有ｐ５３、ｋ－ｒａｓ基因突变３８例（７６．０％）。其中有ｐ５３基因突变２８例（５６．０％）,ｋ－ｒａｓ基因突变１６例（３２．０％）。在１２例出现肝转移者的胆汁中检出有与原发灶相同的点突变基因者１０例     

高危性人乳头瘤病毒与抑癌基因P53在膀胱癌中的表达

用ＰＣＲ法检测５２例膀胱癌组织中人高危性乳头瘤病毒（ＨＰＶ）ＤＮＡ，同时用免疫组化方法检测其Ｐ５３蛋白的过度表达，发现２８例（５３．８４％）ＨＰＶＤＮＡ阳性，１９例（３６．５４％）肿瘤细胞核Ｐ５３染色阳性，二者同时阳性者３例（５．７７％）。ＨＰＶＤＮＡ阳性者多见于高分化、非侵袭性肿瘤中，相反Ｐ５３蛋白阳性则主要表现在低分化、侵袭性肿瘤中。ＨＰＶＤＮＡ和Ｐ５３的表达呈显著性负相关（ｒ＝－０．５７６９）。结果提示：ＨＰＶ感染或Ｐ５３的过度表达与膀胱肿瘤的生物学行为有关，并可将其作为膀胱肿瘤的预后评价指标。     

p53和c—myc异常表达与胃癌细胞多药耐药性关系的研究

应用ＬＳＡＢ免疫组织化学方法研究６７例胃标本中ｐ５３和ｃｍｙｃ的表达与多药耐药性（ＭＤＲ）的关系。结果显示：本组胃癌中ｐ５３阳性３２例，阳性率４７８％；ｃｍｙｃ阳性３７例，阳性率５５２％；Ｐｇｐ阳性３９例，阳性率５８２％。ｐ５３的异常表达与ｍｄｒ１基因表达呈显著正相关（ｒ＝０６３，Ｐ＜００５），而ｃｍｙｃ和ｍｄｒ１的表达无明显相关。提示ｐ５３异常表达可增加ｍｄｒ１基因的表达，从而使胃癌细胞获得ＭＤＲ表型。     

IMMUNOHISTOCHEMICAL DETECTION OF p53 PROTEIN OVEREXPRESSION VERSUS GENE SEQUENCING IN URINARY BLADDER CARCINOMAS

PURPOSE: Mutations of p53 tumor suppressor gene and nuclear accumulation of p53 protein are common in bladder tumors. The prognostic significance of p53 alterations in bladder tumors has not been established. The aim of the present study was to evaluate an immunohistochemical (IHC) method for the routine determination of p53 protein overexpression in human bladder tumors and to determine the relation between nuclear accumulation of p53 with the traditional prognostic indicators and patient survival. MATERIALS AND METHODS: 104 transitional cell carcinomas of the bladder were analyzed simultaneously by immunohistochemistry for p53 protein overexpression and direct DNA sequencing for p53 gene mutations. RESULTS: The overexpression of p53 protein was reported in 30.8% of the cases and mutations of p53 gene in 23.0%. A significant association was observed between p53 alterations established either by IHC or direct DNA sequencing and stage (p<0.0001), grade (p<0.001), vascular invasion (p = 0.0005), DNA ploidy (p = 0.0002) and carcinoma in situ (p<0.0001). The correlation between the p53 gene mutations and p53 nuclear reactivity as detected by IHC was highly significant (p<0.0001). Univariate statistical analysis showed that the expression of p53 was significantly correlated to poor prognosis (p<0.0001). However, in multivariate analysis, only stage was significantly correlated to prognosis (p<0.0001). CONCLUSIONS: The IHC method was highly sensitive and specific and simple to apply for the routine examination of p53 overexpression in bladder tumors. However, overexpression of p53 as determined immunohistochemically, does not appear to have a better predictive prognostic value than stage in bladder tumors.     

p16/p38 MAPK/p53/Wip1通路在乳腺癌发生、发展中的作用及其临床意义

目的 探讨p16/p38 MAPK/p53/Wip1通路在乳腺癌发生、发展中的作用及其临床意义.方法 应用免疫组织化学方法检测70例乳腺癌组织、癌旁组织、20例正常乳腺组织中Wip1、p53、p38 MAPK、p16蛋白的表达,并对Wip1蛋白高表达与p53、p38、p16蛋白表达进行相关分析.结果 3种组织中Wip1蛋白高表达率分别为62.9%(44/70)、2.9%(2/70)、0(0/20).乳腺癌组织比癌旁组织、正常乳腺组织明显升高(P＜0.01).Wip1蛋白高表达与p53、p38、p16蛋白表达呈负相关(P＜0.01,等级相关系数rs分别为-0.529、-0.626、-0.499).结论 p16/p38 MAPK/p53/Wip1是负反馈通路,它可能在乳腺癌发生发展中起重要作用.     

p53 Gene mutations in superficial bladder cancer

OBJECTIVES: To assess the presence of p53 gene mutations in superficial tumors of the urinary bladder (transitional cell carcinoma) and their relationship to classic prognostic factors for cancer recurrence and progression. To analyze the implication of these mutations on the P53 protein structure. MATERIALS AND METHODS: Observational, cross-sectional study of 90 consecutive patients, 60 with superficial transitional cell carcinoma (pTa and pT1) and 30 without neoplastic disease (control group). Samples of bladder tumor and control normal mucosa were analyzed by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) to detect p53 mutations in exons 5-9. Automatic sequencing was used to characterize the mutations and their effect on the P53 protein was analyzed. Bivariate analysis was used to assess the association with other prognostic factors. RESULTS: PCR-SSCP found no mutations in any control group patient, whereas 38.3% of patients with superficial transitional cell carcinoma had one or more mutations in the exons analyzed. Thirty mutations were sequenced; all were point mutations and 86.67% were considered relevant for the P53 structure. A total of 93.3% of the mutations were located in highly conserved regions and 73.3% in mutational hot spots. The highest cell differentiation grades and pT1 stage were associated with a higher incidence of p53 gene mutations. Previous recurrences and other tumor-related histological variables were not associated with a higher percentage of mutations. CONCLUSION: Mutations at p53 did not appear in healthy bladder mucosa and were significantly more frequent in pT1 and high-grade (G-II and G-III) tumors. All mutations detected were point mutations and most caused considerable P53 structural abnormalities, implying major repercussions on P53 function. These data suggest that certain p53 mutations may have prognostic value, even though they were not associated with other classic recurrence and tumor progression parameters. Future analyses of the progress of patients with superficial bladder transitional cell carcinoma and mutated p53 will help clarify this aspect.     

p53 Protein and Gene Alterations in Pathological Stage C Prostate Carcinoma

Purpose

We determined the extent of p53 immunoreactivity in pathological stage C prostate cancer as well as its correlation to tumor grade, substage, recurrence and proliferation rate. To define better the temporal relationship of p53 nuclear reactivity in prostate cancer p53 immunoreactivity was evaluated in all associated prostatic intraepithelial neoplasia lesions.

Materials and Methods

Using immunohistochemistry p53 status and proliferation rate were determined in 96 tumors from patients with pathological stage C prostate cancer. Single strand conformational polymorphism in exons 5 to 8 was used in a subset of specimens to assess the association of p53 nuclear accumulation with mutations in the p53 gene.

Results

p53 Nuclear reactivity was demonstrated in 10 tumors (10.4%), including 6 with high and 4 with low level nuclear reactivity. Of the tumors 86 (89.6%) had no evidence of p53 immunoreactivity. Each of the 6 tumors with high level p53 reactivity had associated areas of prostatic intraepithelial neoplasia that also showed p53 nuclear reactivity. Furthermore, pathological stage C substage (C1, 2 or 3) was significantly associated with p53 nuclear reactivity (p = 0.04). Proliferation rates were correlated with p53 nuclear reactivity (p = 0.09), while there was no association with tumor grade or recurrence. p53 Gene alterations were noted in 2 of the 3 p53 positive tumors versus no alterations in the p53 gene of 3 p53 negative tumors.

Conclusions

p53 Nuclear accumulation is uncommon in pathological stage C prostate cancer and its presence in premalignant prostatic intraepithelial neoplasia lesions suggests that it may be an early event in a subset of prostate cancers.     

变性凝胶电泳和自动DNA序列分析检测结直肠肝转移癌p53基因突变的研究

目的　了解变性梯度凝胶电泳(DGGE)和自动DNA序列分析方法检测肿瘤基因变异及p53基因、p53蛋白在大肠癌发生、转移过程中的动态变化。方法　以DGGE及自动DNA序列分析法检测41例大肠癌原发病灶和肝转移灶p53外显子5～11的基因突变。以免疫组织化学方法检测p53蛋白表达。结果　41例中24例有p53基因突变(62%),其中6例仅在肝转移灶发现p53基因突变,其余均为原发灶、转移灶有一致性的突变。另有3例原发灶即有p53基因突变的病人,在转移灶除保留原有突变外,还出现新增加的突变。在原发、转移灶同时有突变的16例中,14例呈现突变的p53碱基峰和正常峰之比在肝转移灶明显高于大肠癌原发灶(P＜0.001)。p53免疫组织化学染色结果和DGGE、DNA序列分析结果高度一致。但在基因分析呈无义突变的癌灶,免疫组织化学显示p53蛋白的过度表达。结论　在大肠癌肝转移过程中,p53基因突变主要开始于肠癌原发灶,并被保持于转移至肝脏的癌细胞内,在转移灶其含量或含突变型p53癌细胞量明显增加。p53基因突变与p53蛋白过度表达呈正相关关系。DGGE和自动序列分析法只有在于免疫组织化学方法结合使用时,才能对基因改变作出最全面的判断。     

大肠癌细胞p53基因突变与细胞凋亡及细胞倍性的关系

目的　了解ｐ５３ 基因突变、细胞倍性、细胞凋亡三者的关系,探讨突变型ｐ５３ 基因在肿瘤发生中的作用机制。方法　采用 Ｐ Ｃ Ｒ Ｓ Ｓ Ｃ Ｐ（ 单链构象多态性） 检测ｐ５３ 基因突变,用流式细胞仪 Ｆａｃｓｃａｎ 检测大肠癌细胞染色体倍性、凋亡率及其在细胞周期各相中的分布情况。结果　大肠癌标本ｐ５３基因突变率为５０ ％ ;其中突变组肿瘤细胞凋亡率（２２ ．１１ ％ ） 明显低于未突变组细胞凋亡率（４０ ．５７ ％ ） ,（ Ｐ＜ ０ ．０５） ;突变组异倍体细胞百分率（ 占７６ ．９ ％ ） 明显高于未突变组（ 占４６ ．２ ％ ） ,（ Ｐ＜ ０ ．０５） ;异倍体肿瘤细胞凋亡率为３４ ．０ ％ ,二倍体肿瘤细胞凋亡率为４０ ．７ ％ ,二者差异无显著性（ Ｐ＞ ０ ．０５） 。结论　ｐ５３ 基因突变使肿瘤细胞凋亡率下降,进而促进肿瘤发生、发展。ｐ５３ 基因突变多见于异倍体细胞,细胞凋亡率与细胞倍性无关。     

Detection of human papillomavirus DNA and p53 gene mutations in human prostate cancer

The relationship between integration with human papillomavirus (HPV) and p53 gene mutations in tissues of prostate cancer was examined. Tissue samples analyzed were obtained by total prostatectomy (29 stage B cancer cases) and from autopsy (22 endocrine therapy-resistant metastatic disease cases). HPV DNA was detected in 8 of 51 (16%, 5 in stage B and 3 in autopsy cases) by polymerase chain reaction (PCR) using consensus primers on L1 region. Genotypes of HPV were entirely type 16. Structural abnormalities of p53 gene were detected in 7 of the 22 autopsy cases (32%) by PCR-single-strand conformation polymorphism analysis and direct sequencing. No p53 gene mutation was found in stage B cancer cases. Analysis of mutation spectra revealed clear differences between Japanese and Westerners. There was a significant difference in the mutation frequency between stage B and autopsy cases (P < 0.01, Fisher's exact test). One case showed both integration of HPV and p53 gene mutation in different cancer foci. However, the other cases revealed an inverse correlation between the presence of HPV DNA and p53 gene mutations. These data show that p53 genetic alteration is correlated with the progression of prostate cancer, in contrast to the integration of HPV that may occur in a relatively early stage. In conclusion, this study may indicate that either p53 gene mutation or the presence of HPV's oncogenic protein E6 is involved in the development of prostate cancer. © 1996 Wiley-Liss, Inc.     

结直肠癌患者血清p53抗体与肿瘤p53蛋白表达关系的研究

目的　探讨结直肠癌患者血清p5 3抗体与肿瘤组织p5 3蛋白表达的关系 ,评价其能否间接反映p5 3基因的突变。方法　对 13 2例手术治疗的结直肠腺癌患者和 3 6例非肿瘤患者进行血清p5 3抗体定性检测。结直肠癌标本行p5 3蛋白免疫组化染色。结果　结直肠癌组中 5 3例 (4 0 .2 % )血清p5 3抗体阳性 ,而对照组仅 1例 (2 .9% )阳性 (P <0 .0 1)。在 75例p5 3蛋白染色阳性者中 5 1例 (68.0 % )血清p5 3抗体阳性 ;而 5 7例p5 3蛋白染色阴性的患者中 ,仅 2例 (3 .5 % )血清p5 3抗体阳性。结直肠癌血清p5 3抗体阳性率与肿瘤组织p5 3蛋白免疫组化染色阳性反应有明显关系 (P<0 .0 1)。结论　血清p5 3抗体能反映结直肠癌p5 3蛋白的阳性表达 ;检测血清p5 3抗体是间接判定P5 3基因突变的一种可靠而简便的方法。     

p53 mutations in prostate cancer bone metastases suggest that selected p53 mutants in the primary site define foci with metastatic potential

PURPOSE: This study was undertaken to establish the pattern of specific p53 gene mutations in prostate cancer within primary tumors and distant metastases. MATERIALS AND METHODS: We performed polymerase chain reaction-single-strand conformation polymorphism and sequencing analyses of p53 exons 5-8 in DNA extracted from 22 formalin-fixed, paraffin-embedded tissues from 17 patients. Samples from three patients included specimens from primary and metastatic sites (paired specimens). RESULTS: G:C-to-A:T transitions were the most common point mutations (64%). Six (55%) of 11 G:C-to-A:T transitions occurred at CpG dinucleotides in five hot-spot codons (175, 245, 248, 273, and 282). Sequencing analysis of the paired samples revealed that two of the three pairs had the same mutation in both. Sequencing analysis of DNA from a different area of one of the primary tumors revealed a different mutation in the p53 gene. CONCLUSIONS: Our results suggest that specific p53 mutations participate in the progression of human prostate cancer. These findings support those of others that indicate that the primary cancer is heterogeneous and clonal expansion occurs during the progression of clinically detectable prostate cancer. Our data also imply that p53 mutations at the primary site may be predictive of metastases.     

p53上调促凋亡因子在胃癌组织中的表达及其临床意义

目的 研究p53 上调促凋亡因子(p53 up-regulated modulator of apoptosis,PUMA)在胃癌绀织中的表达及其与临床病理学指标的关系.方法 应用组织芯片和免疫组化Envision方法 检测84例胃癌及配对的癌旁正常胃黏膜组织中PUMA和p53蛋白表达;分析PUMA表达与临床病理学指标的关系;分析PUMA表达与p53表达的相关性.结果 PUMA在胃癌组织中的阳性率(33%)低于癌旁正常胃黏膜组织(62%),两者之间筹异有统计学意义,P<0.01;PUMA的表达与肿瘤浸润深度、pTNM分期及预后有关(P<0.05).p53在胃癌组织中的表达(45.2%)显著高于癌旁正常胃黏膜组织(0),两者之间差异有统计学意义,P<0.01;PUMA与p53表达有显著的相关性(P<0.05).结论 PUMA低表达与胃癌浸润深度、胃癌中、晚期及预后不良相关,有可能成为胃癌预后的预测指标.