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     

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。     

ras和p53在甲状腺癌中的表达

目的 探讨ｒａｓ和ｐ５３基因过度表达在甲状腺癌发生中的相互作用,以及甲状腺癌的临床病理标准与ｒａｓ和ｐ５３基因过度表达的关系。方法 采用ＬＳＡＢ免疫组化法检测了ｒａｓ和ｐ５３在８０例甲状腺病变中的表达,其中５４例为甲状腺癌,２６例为良性甲状腺结节性病变。结果 ｒａｓ和ｐ５３在甲状腺癌和良性甲状腺结节性病变中的表达阳性率分别为９０．７％及２３．０％和５５．５％及３０．７％,两者相比较,差异有显著性意     

膀胱癌组织中p53基因突变的研究

作者应用聚合酶链反应－单链构象多态性分析（ＰＣＲ－ＳＳＣＰ）和直接测序等分子生物学技术，分析检测了２１例膀胱移行上皮癌ｐ５３基因５～８外显子。结果，在１４例浅表性膀胱癌中，发现１例（Ｔ＿１期）ｐ５３基因突变；７例浸润性膀胱癌中，４例发现ｐ５３基因突变，其中Ｔ＿２期１例，Ｔ＿３期２例，Ｔ＿４期１例。４例Ｇ＿１级膀胱癌无一例检出ｐ５３突变，５例Ｇ＿２级膀胱癌１例检出ｐ５３突变，１２例Ｇ＿３级肿瘤４例发现ｐ５３基因突变。结果表明：ｐ５３基因突变在膀胱癌的发生发展中起着重要作用。     

肝细胞癌组织中p53基因突变的研究

目的探讨黄曲霉素（ＡＦＢ１）污染严重的南宁地区的肝细胞癌（ＨＣＣ）与ｐ５３基因突变的关系。方法采用聚合酶链反应限制性片段长度多态性分析,检测６５例ＨＣＣ患者的ｐ５３基因第７外显子点突变。结果３８例ＨＣＣ的ｐ５３基因第２４９密码子有集中的点突变,突变频率为５８５％;伴有乙型肝炎病毒阳性者ｐ５３基因突变率５７４％,阴性者６３６％;高度分化、中度分化和低度分化ＨＣＣ的ｐ５３基因突变率分别为１００％、５５２％和８０８％;伴ＨＣＣ转移者ＨＣＣ的ｐ５３基因突变率为８８９％,未转移者为４６８％。结论ＡＦＢ１是产生ｐ５３基因突变热点的主要原因,而乙型肝炎病毒则具有协同作用;ｐ５３基因突变与ＨＣＣ的分化程度及转移有关,并预示着ＨＣＣ的恶性程度,可能是一个具有重要意义的预后指标     

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 …

目的 探讨人乳头瘤病毒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。     

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。     

胃癌组织中p16,p53蛋白的表达及其临床意义

为探讨ｐ１６，ｐ５３蛋白在胃癌发生、发展中的作用及其临床意义。应用免疫组化方法，对６０例胃癌、１８例胃良性疾病及１５例胃正常组织中ｐ１６，ｐ５３蛋白的表达情况进行检测。结果显示：胃癌组织中ｐ１６蛋白的阳性率（２５％）低于胃良性疾病（６１．１１％）及胃正常组织中的阳性率（６３．３３％），有显著性差异（Ｐ＜０．０５）；而胃癌组织中ｐ５３蛋白的阳性率（６１．６７％）高于胃良性疾病（１１．１１％）及胃正常组织（０），差异非常显著（Ｐ＜０．００１）。ｐ１６，ｐ５３蛋白的表达与胃癌的组织学分化、临床分期及淋巴结转移密切相关，与患者年龄、性别、肿瘤位置无关；ｐ１６蛋白表达还与肿瘤大小有关。研究结果表明，ｐ１６蛋白的失表达发生在胃癌发生、发展的晚期阶段，ｐ１６，ｐ５３蛋白可作为判别胃的良、恶性疾病及胃癌患者预后的一个指标。     

胃癌组织中p53蛋白表达的临床意义

目的：观察ｐ５３蛋表达通融作为判断胃癌病的预后指标,为该指标的合理应用提供科学依据。方法 利用免疫组化技术检测１７１例胃癌切除标本内ｐ５３蛋白的表达,分析ｐ５３蛋白表达与临床病理及承后之间的关系,探讨ｐ５３蛋白表达与传统ＴＮＭ分期在预后中的关系。结果 ４３．３％的胃癌组织有ｐ５３蛋白表达,弥漫型胃癌中的ｐ５３蛋白表达率高于肠型胃癌（Ｐ〈０．０５）;有ｐ５３蛋白表达的胃癌组织易于发生洒巴结转移（Ｐ〈     

在胆管癌组织中检测p53,p21WAF1的表达

目的 研究胆管癌中ｐ５３,ｐ２１＾ＷＡＦ１基因表达以及它们之间的关系。以期为找到一种早期诊断及判断胆管癌预后的方法找下一个基础。方法 本研究采用３０例人胆管癌及癌旁组织配对资料,应用原位杂交及免疫组化方法检测、ｐ５３、ｐ２１＾ＷＡＦ１基因表达经们之间的关系。进而分析它们与临床病理资料之间的相关性。结果 原位杂交检测３０例胆管癌中有１４例（１４／３０,４６．７％）呈ｐ５３阳性,１２例（１２／３０,４０．０％）ｐ２１＾ＷＡＦ１阳性。ｐ５３,ｐ２１＾ＷＡＦ１原位杂交结果与胆管癌的临床病理资料无明显相关性。免疫组化检测３０例胆管癌中,Ｐ５３蛋白表达阳性率明显高于无局部淋巴结转移组;临床Ⅲ期的病人Ｐ５３蛋白阳性率明显高于临床Ⅰ和Ⅱ期的病人,并且有Ｐ５３蛋白表达的病人其生存期明显短于无Ｐ５３蛋白表达的病人。ｐ２１＾ＷＡＦ１     

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。     

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.     

P53 Overexpression and Proliferative Potential in Malignant Meningiomas

Summary  Meningiomas are generally benign, but some meningiomas show malignancy with invasion and high recurrence rates. We investigated whether alterations in p53 protein may contribute to malignant progression in meningiomas. Immunostaining for p53 protein was performed on paraffin and frozen sections from 61 patients with different grades of meningiomas using monoclonal antibodies (mAbs) DO-1 and pAb240. Immunoblot analysis was performed to quantitate the amount of p53 protein. Mutations in p53 genes were assessed by single-strand conformational polymorphism (SSCP) analysis. MIB-1 immunostaining was used to detect proliferative potentials of meningiomas. We found an overexpression of p53 protein in all of five cases of anaplastic meningiomas by immunohistochemistry using DO-1 mAb. No p53 positive cells were recognized in atypical meningiomas, and several cells were weakly stained in only two of 52 benign meningiomas.  p53 staining index and immunoblot analysis indicated increasing amounts of p53 protein associated with subsequent recurrences of anaplastic meningiomas. The MIB-1 staining index was positively correlated with tumour grade and p53 protein overexpression. Immunostaining of frozen sections using the mutant-specific mAb pAb240, as well as mutation gene analysis by SSCP, indicate that the overexpressed p53 protein is not a mutant- but wild-type p53 protein. Four atypical meningiomas did not recur after surgical removal and radiation, while 4 anaplastic meningiomas with overexpressed p53 protein recurred repeatedly at short intervals even after radiation. Our results suggest that accumulation of p53 protein associated with highly proliferative potentials is a common and characteristic feature that may indicate malignant biological behaviour in meningiomas.     

p53 overexpression and K-ras gene mutations in primary sclerosing cholangitis-associated biliary tract cancer

Cholangiocarcinoma occurs frequently in patients with primary sclerosing cholangitis (PSC). We evaluated the incidence and prognostic significance of p53 protein overexpression and K-ras gene mutations in patients with biliary tract cancer and PSC. p53 protein expression was determined in specimens from 12 patients with biliary tract cancer, using the antibody, D07. K-ras mutations were detected using DNA sequencing and a mutation ligation assay. Accumulation of p53 protein was detected in 6 of 12 tumors (50%). K-ras mutations were detected in 4 of 12 tumors (33%). Overall survival in patients with p53-negative tumors was significantly longer (P < 0.05) than that in patients with p53-positive (mutant) tumors. Similarly, overall survival was significantly longer (P < 0.05) in the absence of a K-ras mutation than in patients with a tumor containing a K-ras mutation. Mean interval from the time of diagnosis of PSC until the diagnosis of biliary tract cancer was significantly shorter (P < 0.05) in patients with p53 overexpression than in those patients without p53 overexpression (2 versus 47 months). p53 overexpression and K-ras mutations occur commonly in patients with PSC and biliary tract cancer and are associated with a shortened survival. Patients with longstanding PSC are less likely to have these genetic alterations and may have a better prognosis. Received: September 24, 1999 / Accepted: June 6, 2000     

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。     

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、p16在胆囊癌的表达及其临床意义

本研究应用免疫组化ABC法对53例胆囊癌组织及因其他疾病手术切除的正常胆囊组织15例(均经病理证实)进行了p53、p16基因产物的联合检测。结果:p53基因改变发生于胆囊肿瘤的早期,临床Ⅳ期和低分化腺癌的的表达水平分别高于Ⅰ期和高分化腺癌,提示p53的检测可能作为胆囊肿瘤预后及腺癌恶性程度判断的潜在指标。p16表达的改变主要发生在肿瘤的中、晚期,可能与肿瘤的转移和腺癌恶性程度有关,对判断肿瘤转移倾向有一定帮助。联合检测p53、p16可提高对肿瘤的生物学特性的反映。     

Association of p53 tumor suppressor gene with paraclinical and clinical modalities of gliomas patients in Malaysia

Summary Background. Alteration of the tumor suppressor gene p53 is considered to be a critical step in the development of human cancer. Changes in this gene have been detected in a wide range of human tumours, including gliomas. In glioma, the presence of p53 gene alterations has been associated with worse prognosis.Methods. Forty-seven Malaysian adult glioma patients of the Malay race were prospectively studied over a period of 3 years where the presence of p53 mutation using cold-SSCP method and their clinical and paraclinical response were correlated.Findings. Among these glioma patients, p53 mutations were detected in 12 tumors, an incidence rate of 25.5%. Mutations were found in 2 patients of grade II, and 5 patients both in grade III and grade IV. The sequencing results revealed the presence of base-substitutions (7) (58.3%) and frameshifts mutations (5) (41.7%). Of the base-substitutions, 57.1% were transversions and 42.9% were transitions.Interpretation. Our analysis shows that 3 factors were associated with p53 mutations i.e. grade, site and consistency of tumour using univariate analysis although multivariate analysis revealed no positive on predictors of mutation. In conclusion, although p53 genetic alterations are involved in glioma patients in Malaysia, it has no impact on prognosis.     

MDM2/p53 protein expression in the development of colorectal adenocarcinoma

The murine double minutes 2 (MDM2) oncoprotein inhibits p53-mediated tumor suppression. MDM2 has been shown to be overexpressed in sarcomas and more recently was implicated in the pathogenesis of carcinomas. The purpose of this study was to determine the expression pattern of MDM2 in adenomas and colorectal adenocarcinomas and decide whether there is a correlation between MDM2 and p53 protein status. Paraffin-embedded tissues from 52 colorectal cancer (CRC) specimens and their adjacent normal tissue (N-CRC) were studied. In addition, 56 sporadic adenomas were investigated for the immunohistochemical expression of MDM2 and p53 proteins. Immunoreactivity of p53 indicating p53 gene mutation (p53 +) was significantly higher in CRC (44%) compared to adenomas (23.2%) (P <0.01). None of the N-CRC specimens expressed the immunoreactive p53 protein. MDM2 overexpression (MDM2+) was similar in adenomas (30.3%) and CRC (25%), but only 2 (3.8%) of 52 N-CRC specimens showed overexpression of MDM2. In most cases MDM2 expression was associated with negative p53 expression (wild-type p53) in both adenomas (r = 0.59, P <0.001) and CRC (r = 0.69, P <0.0001). No correlation was found between MDM2, p53 expression, and either the histologic grade, nodal stage or morphology of the tumors. There is greater p53 mutation in CRC compared to adenomas and N-CRC. The data indicate that MDM2 is overexpressed in CRC and is significantly associated with wild-type p53 compared to N-CRC specimens from the same patient. The MDM2 expression pattern is similar in adenomas and CRC, which may suggest that MDM2 overexpression is an early event in the progression of CRC. Supported by a grant from The Methodist Hospital Foundation, Houston, Tex. Presented at the Fortieth Annual Meeting of The Society for Surgery of the Alimentary Tract, Orlando, Fla., May 16–19, 1999.