应用PCR—SSCP—EB染色法检测结直肠肿瘤P53基因突变

运用ＰＣＲ－ＳＳＣＰ－ＥＢ染色法对３０例结直肠腺癌和１８例结直肠癌瘤中Ｐ５３基因外显子５、６、７、８突变分别进行检测，结果显示腺癌，腺瘤中Ｐ５３基因突变率分别为６６．７％和３３．３％（Ｐ〈０．０５），高、中、低分化腺癌中Ｐ５３基因突变率分别为３７．５％，６２．５％和８５．７％（Ｐ〈０．０５）。结果提示Ｐ５３基因突变与结直肠腺癌分化程度有关，在结直肠腺瘤向腺癌恶突过程中可能起着重要作用，ＰＣＲ－ＳＳ     

前列腺癌中P53和bcl-2蛋白的表达及意义

应用免疫组化ＬＳＡＢ法，检测１５例良性前列腺增生症（ＢＰＨ）和３５例前列腺癌（Ｐｃａ）组织中Ｐ５３、ｂｃｌ－２蛋白的表达水平。结果发现：（１）ＢＰＨ组与Ｐｃａ组中Ｐ５３、ｂｃｌ－２蛋白的阳性表达率分别为６．７％、１３．３％和３４．３％、４２．９％，显示Ｐｃａ组Ｐ５３、ｂｃｌ－２蛋白阳性率明显高于ＢＰＨ组（分别Ｐ〈０．０５、Ｐ〈０．０５）。（２）Ｐｃａ组中Ｐ５３蛋白阳性１２例，其中Ⅰ、Ⅱ级肿瘤阳性５     

前列腺癌MDR－PGP170与突变P53表达

应用单克隆抗体（ＭＤＲ－Ｃ２１９、ＰＡｂ－ＤＯ－１）免疫组化技术对３８例石蜡包埋的前列腺癌与１５例良性前列腺增生（ＢＰＨ）组织切片中的ＭＤＲ－ＰＧＰ与突变Ｐ５３的表达进行了比较研究。结果显示：在３８例前列腺癌切片中ＭＤＲ－ＰＧＰ与突变Ｐ５３蛋白的阳性表达率为７１．０％和６０．５％，１５例ＢＰＨ组织中仅３例（２０．０％）显示ＭＤＲ－ＰＧＰ阳性染色，突变Ｐ５３在所有ＢＰＨ中未见阳性染色。在高分化、中等分化和低分化前列腺癌中ＭＤＲ－ＰＧＰ的阳性表达率为５３．８％、７８．５％和８１．１％。突变Ｐ５３的阳性表达率为３０．８％、７１．４％和７２．７％。结果提示：（１）ＭＤＲ－ＰＧＰ的过度表达是前列腺癌产生抗药的主要机制；（２）ＭＤＲ－ＰＧＰ与突变Ｐ５３的过度表达可能是前列腺癌细胞适存、转化、增殖与进展的重要方式；（３）突变Ｐ５３的过度表达对ＭＤＲ－１基因可能具有调节与增强效应。     

前列腺增生和前列腺癌雌,孕,雄激素受体研究

采用荧光组织化学法测定５０例良性前列腺增生（ＢＰＨ组）、６例前列腺癌（ＰＣ组）和１２例正常前列腺（Ｎ组）的雌激素受体（ＥＲ）、孕激素受体（ＰＲ）和雄激素受体（ＡＲ）。结果显示ＢＰＨ组和ＰＣ组ＥＲ阳性率高于Ｎ组（Ｐ＜０．０５）。ＢＰＨ组ＡＲ阳性率高于ＰＣ组（Ｐ＜０．０５）。提示前列腺组织中性激素受体（ＳＨＲ）的改变与ＢＰＨ和ＰＣ有密切关系，ＰＣ和ＢＰＨ中ＳＨＲ阳性者适用于内分泌治疗。     

一例前列腺癌P53基因第6外显子中的移码突变

以ＰＣＲ单链构象多态（ＰＣＲＳＳＣＰ）分析及ＰＣＲ直接测序技术，结合免疫组织化学方法，对１例前列腺癌患者治疗前的前列腺穿刺组织及内分泌治疗后复发的手术切除标本中不同部位随机取材的肿瘤组织进行Ｐ５３基因第５，６外显子及Ｐ５３蛋白表达的检测，发现在第６外显子的第２０８密码子存在碱基插入突变（ＧＡＣ→ＧＡＴ），免疫组织化学检测中未发现Ｐ５３蛋白的过量表达。前列腺癌中尚未见有此种Ｐ５３基因突变的文献报道。     

前列腺癌MDR—PGP170与突变P53表达

应用单克隆抗体（ＭＤＲ－Ｃ２１９，ＰＡｂ－ＤＯ－１）免疫组化技术对３８例石蜡包埋的前列腺癌与１５例良性前列腺增生（ＢＰＨ）组织切片中的ＭＤＲ－ＰＧＰ与突变Ｐ５３的表达进行了比较研究。结果显示：在３８例前列腺癌切片中ＭＤＲ－ＰＧＰ与突变Ｐ５３蛋白的阳性表达率７１．０％和６０．５％，１５例ＢＰＨ组织中仅３例（２０．０％）显示ＭＤＲ－ＰＧＰ阳性染色，突变Ｐ５３在所有ＢＰＨ中未见阳性染色。在高分化，中等分     

应用PCR—SSCP银染技术检测结直肠癌P53基因突变

应用ＰＣＲ－ＳＳＣＰ银染技术检测３０例结直肠癌Ｐ５３基因第５、６、７和８外显子的突变情况，结果发现１６例（５３．３３％）发生Ｐ５３基因突变，提示：（１）结直肠癌普遍存在Ｐ５３基因突变。（２）比较传统的同位素标记ＰＣＲ－ＳＳＣＰ技术，ＰＣＲ－ＳＳＣＰ银染技术不仅灵敏度相当，而且简便，快速，低耗，重复性好，尤其适于其样本筛选，可在临床实验室中推广。     

肾细胞癌bc1－2、P53、ER、PR的表达及临床意义

为了更好地选择估价肾细胞癌预后的指标，应用免疫组化技术检测５４例肾细胞癌中ｂｃ１－２、Ｐ５３蛋白及雌、孕激素受体的表达。ｂｃ１－２阳性３８例（７０．４％），Ⅰ、Ⅱ级肿瘤阳性３３例，Ⅲ、Ⅳ级肿瘤阳性５例（Ｐ＜０．０５）；Ｐ５３蛋白阳性１０例（１８．３％），对照组无一例阳性（Ｐ＜０．０５）；ＥＲ、ＰＲ阳性分别为２９例（５７．７％）、３９例（７３．７％）。ｂｃ１－２蛋白表达与ＥＲ、ＰＲ呈正相关（Ｐ＜０．０１，ｒ＝０．４５２），与Ｐ５３蛋白表达呈负相关（Ｐ＜０．０１，ｒ＝－０．４２１）。结果提示：ｂｃ１－２蛋白的表达有助于肾细胞癌预后的评估     

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、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。     

P53 and Ki-67 immunoreactivity in human prostate cancer and benign hyperplasia.

Mutation of the p53 gene is one of the commonest genetic abnormalities found in solid human tumours. This gene is probably concerned with the control of cellular proliferation and in view of this we carried out a study of human prostate cancer and benign prostatic hyperplasia, comparing the expression of mutated p53 with measurement of growth fractions as assessed by staining with Ki-67. A series of 29 patients with prostate cancer (CaP) were compared with 34 men with benign hyperplasia (BPH); 22 of 29 prostate cancers (76%) contained Ki-67 immunoreactivity compared with 10 of 34 (29%) BPH. With respect to p53 staining, significantly more prostate cancers (17%) were stained than BPH (0%). The mean Ki-67 score in cancers positive for p53 (4.3%) was greater than that found in cancers negative for p53 (1.2%), but no statistically significant relationship was found between tumour grade and Ki-67 staining. The use of Ki-67 and p53 staining may allow identification of tumours with a higher rate of cell growth and may permit development of prognostic factors.     

P53,Rb抑癌基因突变产物在前列腺癌中的表达

用Ｐ５３和Ｒｂ，通过免疫金－银染色方法，对３０例前列腺癌及２０例前列腺增生症进行检测。结果显示：前列腺癌Ｐ５３、Ｒｂｃ１５的阳性率分别为６０．０％和５０％，前列腺增生症仅有１０％和１５％，二者之间有显著性差异。前列腺正常组织阳性率Ｐ５３ 和Ｒｂ为０，与肿瘤区比较Ｐ＜０．００１；３０例前列腺癌中，有１０例Ｐ５３和Ｒｂ同为阳性，Ｐ５３在低分化和未分化癌中阳性表达明显高于高分化腺癌（Ｐ＜０．０１）。     

p53, c-erbB-2 and the epidermal growth factor receptor in the benign and malignant prostate.

Expression of the p53, the epidermal growth factor receptor (c-erbB-1) and c-erbB-2 protein was studied in 34 men with benign prostatic hyperplasia and 29 men with locally advanced prostate cancer by means of an immuno-histochemical method. Strong staining for p53 was found in five of 29 prostate cancers (17%; mean 21% +/- 7% of malignant cells stained in the positive tumours), but no staining was found in benign prostatic hyperplasia (p less than 0.05). On the other hand, the epithelium in benign glands was stained positively for c-erbB-2 in 18% (6/34) and for the epidermal growth factor receptor in 88% (30/34); whereas malignant epithelium stained strongly for c-erbB-2 in 21% (6/29) and for the epidermal growth factor receptor in only 17% (5/29). Prostate cancer was associated with a significant decrease in epidermal growth factor receptor staining (p less than 0.0001) and a significant increase in p53 staining (p less than 0.05). Most of the tumours were advanced and no significant relationship was observed between tumour stage and grade and expression of p53, the epidermal growth factor receptor or c-erbB-2. These findings demonstrate that altered expression of the epidermal growth factor receptor and p53 protein occurs in prostate cancer, but were not associated with other features of prognostic importance such as stage or grade.     

良、恶性前列腺病变组织P53、C-erbB-2和EGF-R表达及其临床意义

目的 探讨P_(53)、C-erbB-2和EGF-R在良、恶性前列腺病变组织中的表达差异及其临床意义。方法 采用免疫组织化学ABC法,分别对34例良性前列腺增生(BPH)、29例前列腺癌(Pca)组织中P_(53)、C-erbB-2和EGF-R表达进行研究。结果 在29例Pca中P~(53)阳性染色5例,阳性率为17％(5/29),而在良性前列腺组织中则无一例阳性染色(P<0.05)。在良性前列腺增生征中C-erbB-2和EGF-R阳性表达率分别为18％(6/34)和88％(30/34);而在前列腺癌中C-crbB-2和EGF-R的阳性率分别为21％(6/29)和17％(5/29),前列腺癌分期和分级与P_(53)、C-erbB-2和EGF-R的表达之间没有明显的相关性(P>O.05)。结论 尽管P_(53)、EGF-R在良性,恶性前列腺病变之间表达有差异(P<0.05),但是他们与肿瘤的分期和分级等预后因素无关。     

P504S immunohistochemical detection in 405 prostatic specimens including 376 18-gauge needle biopsies

P504S is a recently described, prostate cancer-specific gene that encodes a protein involved in the beta-oxidation of branched chain fatty acids. A recent study has shown that immunohistochemical detection of P504S gene product is a sensitive and specific marker of prostatic carcinoma in formalin-fixed, paraffin-embedded tissues. We performed a detailed analysis of P504S protein expression in a large series of prostate and bladder specimens with special emphasis on staining in specific morphologic patterns of prostatic adenocarcinoma, posthormonal and radiation therapy cases, and invasive urothelial carcinoma. A total of 366 prostate needle core biopsies from 124 patients with prostate cancer, 10 biopsies from 2 patients without prostate cancer, 28 prostatectomy specimens (16 with specific morphologic patterns, 7 posthormonal therapy and 5 postradiation therapy specimens), 5 bladder specimens with invasive urothelial carcinoma, and a single transurethral resection specimen from a patient with hormonally treated prostate cancer and invasive urothelial carcinoma were stained with P504S monoclonal antibody at a 1:250 dilution using standard heat-induced epitope retrieval and avidin-biotin technique. Extent (0, no staining; 1+, 1-10% staining; 2+, 11-50% staining; 3+, > or =51% staining) and location (luminal, subluminal, and diffuse cytoplasmic) of immunoreactivity in carcinoma and benign tissues were recorded. A total of 153 of 186 biopsies (82%) with prostatic adenocarcinoma stained for P504S. Pseudohyperplastic, atrophic, ductal, and mucinous prostatic carcinomas stained similarly, as did cases treated with hormone or radiotherapy. In 81 of 377 (21%) foci of benign prostatic tissue there was staining that was almost always focal, faint, and noncircumferential. Seminal vesicles did not stain for P504S. Five of six (83%) specimens with invasive urothelial carcinoma had 2+ staining and one case had focal staining. We conclude that immunohistochemistry for P504S has potential utility in the diagnosis of prostate cancer, including those treated by hormones and radiation. Circumferential luminal to subluminal and diffuse cytoplasmic staining is the most specific staining pattern for prostatic carcinoma and is almost never associated with benign prostatic tissue. However, a negative P504S immunostain does not automatically rule out prostate cancer, as 18% of cases were negative. Additionally, occasional benign glands, high-grade prostatic intraepithelial neoplasia, atypical adenomatous hyperplasia, and urothelial carcinoma may express P504S. Therefore, we think that P504S is best used only in conjunction with strict light microscopic correlation and preferably with high molecular weight cytokeratin immunostaining.     

前列腺癌中P53的表达及其临床意义

目的:探讨P53在前列腺癌中的表达及其临床意义。方法:应用免疫组化Elivision法,检测45例前列腺癌和10例前列腺增生组织标本P53的表达,分析P53表达与前列腺癌分期、分级、前列腺特异性抗原(PSA)水平、内分泌治疗效果及预后的关系。结果:P53在前列腺癌和前列腺增生中阳性表达率分别为51.1%和10.0%(P<0.05);D期和A~C期前列腺癌中阳性表达率分别为70.0%和25.0%(P<0.05);Gleason评分≤7分和>7分前列腺癌中阳性表达率分别为14.3%和56.7%(P<0.05);PSA≤10μg/L和>10μg/L前列腺癌中阳性表达率分别为20.0%和60.0%(P>0.05);在内分泌治疗有效和无效前列腺癌中阳性表达率分别为25.0%和72.3%(P<0.05);LogRank检验分析P53阴性表达前列腺癌患者的生存时间明显长于P53阳性表达前列腺癌患者(P<0.05)。结论:P53表达可以作为判定前列腺癌预后的分子标记,同时可以预测内分泌治疗的效果。     

Mutations within the tumour suppressor gene p53 are not confined to a late event in prostate cancer progression. a review of the evidence

Mutations in the p53 tumour suppressor gene are generally believed to be a late event in the progression of prostate cancer, and are associated with androgen independence, metastasis, and a worse prognosis. In this review, we examine the current literature available on p53 mutations and focus on stages A (T1) and B (T2) of prostate cancer. We report here that p53 mutations can be found in approximately one third of prostate cancers that are clinically localized to the prostate. In addition, high levels of p53 mutation are found in normal prostate tissue of prostate cancer patients, prostatic intraepithelial neoplasia, and benign prostatic hyperplasia. The limitations of techniques used to determine p53 mutations are discussed, as well as other modes of p53 loss in early stage prostate cancer.     

肿瘤标记物P504s在前列腺癌中的表达及应用价值

目的探讨肿瘤标记物P504s在不同前列腺组织中的表达情况,及在诊断前列腺癌中的应用价值。方法采用免疫组化二步法,观察P504s和基底细胞标记物p63在不同前列腺病变组织中的表达情况。结果①P504s表达在前列腺癌根治标本、高等级上皮内瘤标本、前列腺增生标本中的阳性率分别为91.4%（32/35）、87.0%（20/23）和0（0/30）,良恶性组织的差别有显著性,在前列腺癌穿刺标本中阳性率85.7%（72/84）。p63染色在全部前列腺癌中呈阴性,在高等级上皮内瘤中为不连续或连续的阳性染色,在前列腺增生中为连续的阳性染色。②P504s表达与前列腺癌的分化程度、激素治疗、转移有关。结论P504s是一个特异的前列腺癌标记物,结合p63在鉴别诊断前列腺癌中有重要的临床应用价值。     

同源框基因NKX3.1在前列腺癌组织中的表达及意义

目的　探讨同源框基因NKX3.1在前列腺癌中的表达意义。　方法　采用半定量RT PCR检测前列腺癌、良性前列腺、非前列腺组织标本NKX3.1mRNA表达状况并对其扩增产物进行测序。　结果　 76例前列腺组织中 ,NKX3.1表达 75例 ,表达率 98.7%。 96例非前列腺组织标本中 ,除 2例睾丸组织、1例乳腺组织有NKX3.1表达外 ,肾、膀胱、肝、回肠、脂肪、皮肤组织无 1例表达。前列腺癌组织NKX3.1表达量明显增高 ,良性前列腺增生组织表达量明显减弱 (P <0 .0 1) ;晚期前列腺癌组织NKX3.1表达明显高于早期前列腺癌 (P <0 .0 5 ) ;低分化前列腺癌表达高于中高分化前列腺癌 (P <0 .0 5 ) ;激素依赖性前列腺癌表达高于激素非依赖性前列腺癌 (P <0 .0 1)。NKX3.1表达高低与前列腺体积和血清总PSA浓度无关 (P >0 .0 5 )。　结论　NKX3.1基因具有前列腺组织特异性 ,其表达状况与前列腺癌分期、分级相关。NKX3.1表达对于判断前列腺癌病理生物学特性和前列腺癌治疗有重要意义。     

Expression of alpha-Methylacyl-CoA racemase (P504S) in atypical adenomatous hyperplasia of the prostate

Atypical adenomatous hyperplasia (AAH) of the prostate, also known as adenosis, is characterized by a proliferation of prostatic glands with abnormal architectural patterns, but without significant cytologic atypia. In some cases it may be difficult to distinguish AAH from prostatic carcinoma. Additionally, it is not clear whether AAH is a precursor lesion of prostatic adenocarcinoma. P504S, a protein highly expressed in prostatic adenocarcinoma, has been recently shown to be a marker of prostate cancer. The goal of this study is to examine the expression of P504S in AAH by immunohistochemistry. A total of 80 prostate specimens, including 40 cases of AAH (prostatectomy N = 30, biopsy N = 6, transurethral resection N = 4), 20 cases of prostatic adenocarcinomas, and 20 cases of benign prostatic hyperplasia, were studied. Immunohistochemistry for a prostate cancer marker alpha-methylacyl-CoA racemase (P504S) and a basal cell-specific marker 34betaE12 was performed in all the cases. The 34betaE12 stain confirmed the presence of patchy basal cells in all 40 cases of AAH. P504S was undetectable in the majority of AAHs (33 of 40, 82.5%), focally expressed in four of 40 (10.0%), or diffusely positive only in three of 40 (7.5%) cases of AAH. Interestingly, two of seven P504S-positive AAHs were found adjacent to adenocarcinoma. In contrast, all benign prostatic hyperplasias (20 of 20, 100%) were negative for P504S, and all 20 cases of prostatic carcinomas (100%) showed a diffuse P504S staining pattern. These findings suggest that AAH is a heterogenous entity. The biologic significance of P504S expression in a small subset of AAH remains to be determined. Because most cases of AAH are negative for P504S, immunostaining of P504S is also of diagnostic value in distinguishing the majority of AAHs from prostatic adenocarcinoma.