P－糖蛋白在非小细胞肺癌中的表达

采用生物素－亲和素过氧化物酶复合物法，以抗多药抗药性（ＭＤＲ）Ｐ－糖蛋白（Ｐ－ｇｐ）单克隆抗体４Ｅ＿３，检测Ｐ－ｇｐ在２４例非小细胞肺癌中的表达。在１４例腺癌中，Ｐ－ｇｐ表达阳性者１３例，阳性率为９２．９％，１０例鳞癌中，Ｐ－ｇｐ表达阳性者３例，阳性率为３０．０％。腺癌中Ｐ－ｇｐ表达阳性率显著高于鳞癌，提示Ｐ－ｇｐ表达是非小细胞肺癌产生内在抗药性的机制之一。     

p53 Protein Accumulation in Non-Invasive Lesions Surrounding p53 Mutation Positive Invasive Breast Cancers

p53 mutation is a common event in sporadic breast cancer being found in 15–50% of invasive carcinomas. The purpose of this study was to determine the earliest histologic stage at which p53 mutation could be detected with a widely used anti-p53 antibody (DO7, Novocastra) which recognizes both wild type and mutant forms. p53 expression was assessed immunohistochemically in 12 primary breast carcinomas with known p53 mutations and in all pre-malignant epithelial lesions surrounding these invasive cancers. Strong p53 nuclear staining was found in all of the tumors known to have missense mutations and none of the tumors with truncation mutations. In cases with intense staining in the invasive carcinoma, a similar quality of staining was also seen in all areas of DCIS (ductal carcinoma in situ) and was representative of missense p53 mutations. Lighter nuclear staining intensity was observed in up to 40% of cells in areas of hyperplasia and in up to 30% of normal breast lobules irrespective of the type of mutation found in the invasive carcinoma. This weak staining was not specific to mutated p53 and may indicate increased amounts of normal p53 protein.We conclude that p53 inactivation occurs prior to invasion in breast carcinogenesis, with mutations being uniformly identified in DCIS associated with p53-mutated invasive carcinomas. In contrast, there is no evidence that epithelial hyperplasia or epithelial cells of the terminal duct lobular unit harbor the same mutations as their associated invasive carcinoma.     

p53 Accumulation in Colorectal Cancer with Hepatic Metastasis

The prevalence of immunoreactive p53 and argyrophilic nucleolar organizer region (AgNOR) numbers were compared between colorectal cancers with (n=44) and without (n=51) hepatic metastasis for at least 5 years. At the same time, the distribution of p53-positive cells in primary, metastatic, and xenografted tumors from the same individuals were studied. Overall, p53 positivity was found more frequently in the cases with hepatic metastasis than in non-metastatic controls, regardless of the distribution pattern ( P <0.05), whereas AgNOR counts were not different between the two groups. Significant heterogeneity in the distribution of p53 immunoreactivity was noted in both the primary and metastatic lesions. The intratumor distribution patterns of p53 immunoreactive cells in the primary (n=33), metastatic (n=33), and xenografted (n=7) tumors of the same individuals were consistent in the majority of cases. There were a few cases in which the p53 immunoreactive cells were more dominant in the metastatic tumor cells. Our observations suggest that p53 accumulation in colorectal cancer is associated with increased risk for hepatic metastasis, while cell proliferation as represented by AgNOR numbers is not. In addition, heterogeneity of abnormal p53 accumulation in the tumor is maintained during the course of metastasis and even after implantation in nude mouse. p53-Immunoreactive cells in the population of colorectal cancer cells do not necessarily have higher metastasizing potential.     

Bortezomib activity and in vitro interactions with anthracyclines and cytarabine in acute myeloid leukemia cells are independent of multidrug resistance mechanisms and p53 status

Purpose The proteasome inhibitor bortezomib may be effective in combination with cytarabine and anthracyclines in the treatment of acute myeloid leukemia (AML) by virtue of targeting aberrantly activated NF-κB in AML stem cells. We tested whether bortezomib cytotoxicity is affected by multidrug resistance (MDR) proteins expressed in AML cells. We also tested whether bortezomib interactions with cytarabine and anthracyclines are affected by p53, because proteasome inhibition stabilizes p53 and may thus cause cell cycle arrest. Experimental design Bortezomib sensitivity of cell lines overexpressing P-glycoprotein, multidrug resistance protein-1, breast cancer resistance protein and lung resistance protein was studied in the presence and absence of established modulators of these transport proteins. Drug interactions during simultaneous and sequential exposure to bortezomib and anthracyclines or cytarabine in diverse ratios were evaluated by isobologram and combination index analyses in AML cell lines with wild type and inactive p53 and were correlated with cell cycle perturbations induced by bortezomib. Results Of the MDR mechanisms studied, only P-glycoprotein conferred resistance to bortezomib, and resistance was only twofold. Interactions between bortezomib and anthracylines and cytarabine changed from antagonistic to additive or synergistic with increasing drug activity levels and were not affected by p53 status. Conclusions MDR proteins and p53 do not affect bortezomib cytotoxicity or in vitro interactions with anthracyclines or cytarabine, but these interactions are concentration-dependent, and this concentration-dependency should be considered in the design of combination regimens.     

乳腺增生病p53基因第8外显子突变及p53蛋白表达

目的：探讨p53基因在乳腺癌发生早期的作用。方法：用免疫组化检测36例乳腺单纯增生，31例不典型增生，30例乳腺癌组织中p53蛋白表达，用PCR－RFLP检测p53基因第8外显子第278密码子突变。结果：乳腺单纯性增生，不典型增生和乳腺癌中p53蛋白表达率分别为0，22．6％，46．7％。p53基因第8外显子的突变率分别为0，3．2％，10．0％，均为杂合子突变。结论：乳腺癌不典型增生中存在p53蛋白表达和该基因第8外显子突变，该突变可能在乳腺增生病向乳腺癌进展过程中起一定作用。     

Nuclear Accumulation of p53 Protein Correlates with Mutations in the p53 Gene on Archival Paraffin-embedded Tissues of Human Breast Cancer

Fifty invasive ductal carcinomas of the breast were analyzed by immunohistochemistry, polymerase chain reaction-single strand conformationl polymorphism (PCR-SSCP) and direct sequencing after microdissection of conventional formalin-fixed, paraffin-embedded tissues. A highly significant association between the presence of p53 gene mutation and nuclear accumulation of p53 protein was found ( P < 0,0001). Of 13 tumors that demonstrated p53 gene mutations, 11 (84.6%) showed nuclear accumulation of p53 protein. However, of 37 tumors in which gene mutations were not detected, only 5 (13.5%) showed nuclear accumulation of p53 protein. There was a statistically significant association between the nuclear accumulation of p53 protein and a higher histological grade ( P < 0.001) or mitotic index ( P <0.01). In addition, gene mutations had a statistically significant association with a higher histological grade ( P <0.05) or mitotic index ( P > 0.0001). Therefore, p53 abnormalities might be associated with an aggressive phenotype in breast cancer. We conclude that the immunohistochem-ical detection of nuclear p53 protein accumulation is highly associated with p53 gene mutations in archival paraffin-embedded tissues, and that this method is useful for rapid screening of p53 abnormalities.     

High Incidence of p53 Gene Mutation in Human Ovarian Cancer and Its Association with Nuclear Accumulation of p53 Protein and Tumor DNA Aneuploidy

Using the poylmerase chain reaction and single-strand conformation polymorphism analysis, p53 gene mutations were examined in 24 cases of ovarian tumor including 14 ovarian carcinomas and 2 borderline cases of common epithelial type, 7 germ cell tumors, and one stromal tumor. Abnormal bands indicating mutations were detected in 12 (50%) of the cases examined, being present most frequently in common "epithelial" ovarian carcinoma (71%, 10/14). One case each of squamous cell carcinoma originating in a dermoid cyst and anaplastic dysgerminoma were positive for mutation. Direct sequencing confirmed 12 mutations and revealed G→A and G→C nucleotide changes in 5 and 3 cases (42% and 25%), respectively. The mutation was localized at the CpG site of the gene in 3 cases. Immunohistochemical examination of p53 protein in 21 cases and DNA flow-cytometrical analysis in 17 cases were also performed. Nuclear accumulation of the p53 protein and DNA aneuploidy pattern were detected in 11 (52%) and 9 (53%) cases, respectively. These were significantly correlated with p53 gene mutation ( P <0.01 and P <0.05, respectively; Fisher's exact test). Neither mutation of the p53 gene, nuclear accumulation of p53 protein nor DNA aneuploidy was detected in borderline cases of common "epithelial" type, typical dysgerminoma and immature teratoma. These results suggest that p53 gene mutation, nuclear accumulation of the protein and the DNA aneuploidy pattern are events occurring almost simultaneously in the progression of ovarian tumors, and that p53 abnormalities seem to be correlated with a high grade of malignancy.     

抑癌基因p53在胃癌组织的表达及临床意义

目的：探讨ｐ５３基因在胃癌的表达与临床、病理因素及预后的关系,探索ｐ５３基因异常表达在胃癌发生过程中的作用。方法：使用抗ｐ５３蛋白单克隆抗体ＤＯ－７,对９９例人胃癌组织ｐ５３的表达进行免疫组化研究。结果：４８例胃癌组织ｐ５３基因表达阳性（４９％）。ｐ５３基因表达与胃癌的大体类型、分化工、淋巴结转移等指标均无明显关系。在１０例不典型增生的癌旁组织中,２例呈阳性反应;在４例有肠上皮化生的癌旁组织中及其     

大肠癌p53抑癌基因突变与组织类型,Dukes‘分期的关系

用ＰＣＲ－ＳＳＣＰ方法对３０例大肠癌组织ｐ５３基因突变状况进行了检测。结果表明：３０例大肠癌ｐ５３基因的突变率为２０％（６／３０），其中１例在第４外显子，５例在第５外显子，第２、３外显子未检出突变。高分化和中等分化肠癌ｐ５３基因突变率均为１／８（１２．５％），低分化为３／１０（３０．０％），未分化为１／４（２５．０％）；在Ｄｕｋｅｓ分期中的突变率分别为：Ｂ２期为１／１０（１０．０％），Ｃ１期为２／１２（１６．７％），Ｄ期为３／８（３７．５％）。结果提示ｐ５３基因突变与Ｄｕｃｋｅｓ分期有关，且肠癌的ｐ５３基因突变很少发生在第２、３外显子     

大肠癌组织中nm23和p53蛋白的表达及其意义

目的 探讨ｎｍ２３和ｐ５３蛋白在大肠癌组织中的表达及其意义。方法 采用免疫组化ＬＳＡＢ法检测ｎｍ２３和ｐ５３蛋白在５６例大肠癌组织中的表达情况。结果 大肠癌组织中ｎｍ２３蛋白和ｐ５３蛋白表达率分别为２７．７％（１８／６５）和５２．３％（３４／６５）,两者的阳性表达呈负相关（γ＝－０．３１,Ｐ〈０．０５）;ｎｍ２３蛋白和ｐ５３高表达均与大肠癌淋巴结转移、Ｄｕｋｅ’ｓ分期及预后有关,与组织分化程度无关     

Absence of a Mutation of the p21/WAF1 Gene in Human Lung and Pancreatic Cancers

The expression of a negative regulator of the cell cycle, p21^WAF1 protein, is trans-activated hy wild-type p53, but not by the mutant protein. Therefore, mutations of the p 53 and WAF1 genes may be complementary. We examined DNAs from 70 human primary lung (63 of NSCLC and 7 of SCLC) and 24 pancreatic cancers (19 primary cancers and 5 cell lines) for mutations of the WAF1 gene. No mutations were detected in any samples examined, regardless of the mutational state of the p 53 gene. The results suggested that aberrations of the coding sequence of the WAF1 gene are not associated with carcinogenesis in lung and pancreatic cancers.     

散发性结直肠癌、腺瘤组织中hMLH1和突变型p53表达

[目的]探讨hMLH1和p53基因在散发性结直肠癌发生机制中所起的作用。[方法]采用PV-6000二步法免疫组化检测技术对60例散发性结直肠癌，45例大肠腺瘤和26例正常大肠黏膜石蜡切片进行hMLH1和p53表达的检测。[结果]（1）60例散发性结直肠癌hMLH1蛋白阴性表达和p53蛋白阳性表达分别为11例（18．3％）和32例（53.3％），hMLH1蛋白阴性表达和p53蛋白阳性表达与患者年龄、肿瘤部位、组织学类型和分化程度密切相关（P〈0．05或P〈0．01），而与患者性别、肿瘤大体类型、肿块大小、浸润深度、淋巴结及远处转移与否和患者的Duke’s分期均无显著相关性（P〉0．05）。（2）大肠腺瘤患者p53蛋白阳性表达与患者腺瘤部位、组织学类型和不典型增生程度密切相关（P〈0．05），而与患者性别、年龄无显著相关性（P〉0．05）。（3）散发性结直肠癌中hMLH1蛋白阴性表达组p53蛋白阳性表达率（18.2％，2/11）低于阳性表达组（61．2％，30／49）（P〈O．05）。[结论]（1）大部分散发性结直肠癌是沿染色体不稳定途径发生，其中抑癌基因p53的突变起重要作用。同时，一定比例的散发性结直肠癌中存在错配修复基因的缺陷，并具有相对特殊的临床病理特征。（2）大肠腺瘤组织中有一定比例的hMLH1蛋白阴性表达和p53蛋白阳性表达，提示hMLH1和p53基因的突变可能是散发性结直肠癌发生的早期事件之一。     

Allelic Frequency of p53 Gene Codon 72 Polymorphism in Urologic Cancers

Alterations in the p53 tumor suppressor gene appear to be important in the development of many human tumors. The wild-type p53 gene has a polymorphism at codon 72 that presents the arginine (CGC) or proline (CCC) genotype, which recently has been reported to be associated with genetically determined susceptibility to smoking-related lung cancers. To determine whether this p53 genotype influences individual risk of urologic cancer and/or its progression, we used polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis to assay the allelic frequencies of this polymorphism in 85 renal cell carcinoma patients, 151 urothelial cancer patients, 33 testicular cancer patients, 28 prostatic cancer patients and 56 patients without neoplastic disease. The allelic distributions of the three genotypes (Arg/Arg, Arg/Pro, Pro/Pro) in patients with renal cell carcinoma (29.4%, 55.3%, 15.3%), urothelial cancers (45.7%, 39.7%, 14.6%), testicular cancer (45.4%, 48.5%, 6.1%) or prostate cancer (42,9%, 50.0%, 7.1%) did not differ significantly from those in the normal controls. However, Pro/Pro genotype in renal cell carcinoma and urothelial cancer (smoking-related cancers) was more frequent than that in prostate cancer and testicular cancer (smoking-unrelated cancers) with borderline significance (P=0.0881). There was no particular correlation between frequency of the three genotypes and grade or stage of each type of tumor. The association of genetic predisposition to urologic cancers with p53 gene codon 72 polymorphism is not so clear as the previous study of Japanese lung cancer patients, but this polymorphism may play some role in urothelial cancers and renal cell carcinoma, in which smoking is an epidemiological risk factor.     

肺癌患者GSTM1基因型与p53基因突变的相关关系研究

目的:探索肺癌患者Ⅱ相代谢酶主要亚型GSTM1的基因型与p53基因突变的关系.方法:应用双重PCR和PCR-SSCP(单链构像多态性分析)技术,检测了63例肺癌患者和47例健康对照中GSTM1基因缺失及肺癌组织p53基因突变的情况,并分析其相关关系.结果:肺癌组GSTM1基因缺失率为71.4%(45/63),对照组为51.1%(24/47),差异有显著统计学意义,OR为2.40(95%CI为1.09-5.29).p53基因突变率在63例肺癌组织中为49.2%(31/63),在15例对照组织中仅为6.6%(1/18).58.1%肺癌病例同时存在p53基因突变和缺失GSTM1基因(P＜0.05).结论:GSTM1基因缺失与p53基因突变相关,GSTM1基因缺失可能增加p53基因突变的几率,从而导致肺癌患病危险性的增加.     

Alteration of p53 Clonality Accompanying Colorectal Cancer Progression

The aim of this study was to clarify whether or not the status of gene alteration is heterogeneous in intramucosal carcinoma and homogeneous within invasive carcinoma. We selected 10 colorectal carcinoma cases (1 mucosal, 5 submucosal and 4 advanced carcinomas including 2 cases with lymph node metastasis) and analyzed the p53 gene sequence. Six colorectal cancers in this study showed heterogeneity in p53 mutations in cells from the intramucosal part. In the invasive part of a carcinoma, p53 mutation status was homogeneous intratumorally in all cases. These data indicate that, in regard to p53 gene alterations, colorectal cancers can be composed of various subclones when limited to the mucosa, but clonal selection occurs when one of these subclones commences invasion to the submucosa, generating a monoclonal invasive carcinoma.     

乳腺癌中抗癌基因p53突变及其临床预后

本研究以ｐ５３ｃＤＮＡ为探针，检测２９例散发性乳腺癌中的ｐ５３基因突变。２９例乳腺癌中有２０例乳腺癌有ｐ５３基因杂合性缺失（ＬＯＨ），缺失率为６９％。比较不同分期、肿瘤大小及ＥＲ状态的乳腺癌，ｐ５３基因缺失率无显著性差异（Ｐ＞０．０５）；而不同腋淋巴结状态的乳腺癌，ｐ５３基因缺失率有显著性差异（Ｐ＜０．０５）。上述结果表明ｐ５３基因突变是乳腺癌进程中的早期事件；ｐ５３基因的突变可能预示肿瘤具有较强的侵袭能力。     

大肠癌MDR—1与p53表达关系及其意义

目的探讨大肠癌中多药耐药基因ＭＤＲ１与ｐ５３表达之间的关系。方法利用免疫组化研究６０例大肠癌及２０例大肠腺瘤ＭＤＲ－１,ｐ５３的表达。结果显示６０例大肠癌切片中ＭＤＲ－１与ｐ５３的阳性表达率为５７％和３７％,１０例正常大肠组织和２０例大肠腺瘤仅２０％显示ＭＤＲ－１阳性表达,ｐ５３在正常大肠组织,管状腺瘤中全部阴性,仅一例有不典型增生的绒毛状腺瘤为阳性表达。同时显示ｐ５３在不同分化的大肠癌中与ＭＤＲ表达呈正相关。结论１）通过检测ＤＭＲ－１可以对大肠癌病人进行化疗敏感性预测。２）ｐ５３过度表达对ＭＤＲ－１基因可能具有诱导和增加效应。３）ｐ５３和ＭＤＲ－１一起可用作判断病人预后,生物学行为的一个有用参数。     

p53 Status in Multiple Human Urothelial Cancers: Assessment for Clonality by the Yeast p53 Functional Assay in Combination with p53 Immunohistochemistry

Multifocal synchronous or metachronous tumor development is a common observation in human urothelial cancer cases. However, the underlying mechanism has remained obscure. We have employed a new tool to investigate the p53 gene status, the yeast p53 functional assay, in combination with immunohistochemistry in a total of 50 tumor samples from 32 cases with urothelial cancers, including 8 with multiple synchronous tumor development and 2 demonstrating metachronous tumors. p53 mutations were found in 13 cases (9 with missense mutations, 3 with deletion, 1 with splicing mutation) by the yeast p53 functional assay. p53 protein overexpression was seen in all 9 cases with missense mutations, but in only one of the 4 cases with nonsense mutations. Two tumors without p53 mutation also showed positive p53 immunoreactivity. Overall, p53 abnormalities including mutations and/or protein overexpression were found in 15 (47%) cases. p53 abnormalities were significantly more frequent in non-papillary and in high grade tumors. Loss of the wild type allele in addition to a p53 mutation was suggested in 8 of the 15 (53%) cases. All 4 cases with mutations in multiple synchronous tumors had identical p53 mutations in the separate urothelial cancers, strongly suggestive of monoclonality. The one case with multiple metachronous tumors, in contrast, was characterized by variation in the p53 status, indicative of different clonal origins. In conclusion, combined assessment for p53 status as used here (yeast p53 functional assay plus immunohistochemistry) may provide insights into the molecular mechanisms of urothelial carcinogenesis.     

乳腺癌易感癌基因BRCA1与p53的免疫组化研究

目的：探讨ＢＲＣＡ１在家族性乳腺癌中的作用及其与ｐ５３的相关性。方法：对家族性乳腺癌手术标本及非家族性乳腺癌手术标本共８４例ＢＲＣＡ１及ｐ５３蛋折免疫组化（Ｓ－Ｐ法）检测。结果：ＢＲＣＡ１在家族乳腺癌中及非家族性乳腺中的阳性率分别为６１．９％和１１．９％,ｐ５３在家族性乳腺癌及非家族乳腺癌中的阳性率分别为４０．５％和２６．２％,而在家族性乳腺癌ＢＲＣＡ１阳性病例中有５７．７％的病例ｐ５３阳性,在非