大肠腺癌P53基因cDNA突变与突型P53基因蛋白表达的

探讨大肠腺癌突变型Ｐ５３基因蛋白表达与Ｐ５３基因ｃＤＮＡ突变间的相互关系和意义。方法对１００例新鲜大肠腺癌组织采用ＰＡｂ２４０单克隆抗体免疫组织化学染色检测突变型Ｐ５３基因蛋白表达、ＲＴ－ＰＣＲ－ＳＳＣＰ检测Ｐ５３基因ｃＤＮＡ突变，并比较它们间相互关系。     

p53基因蛋白在大肠腺癌中表达的病理学意义

ｐ５３基因蛋白在大肠腺癌中表达的病理学意义＊１季晨阳２季丹３ＤｕｎｃａｎＲＳｍｉｔｈ３Ｈａｋ－ＳｕＧｏｈ野生型ｐ５３基因蛋白是抑癌基因蛋白，可以调控细胞生长、分化和ＤＮＡ修复并促进凋亡〔１，２〕。在发生肿瘤时，ｐ５３基因失活（突变、杂合性丢失），不但...     

用非同位素PCR－SSCP方法检测石蜡包埋乳腺癌p53基因点突变

应用一非同位素ＰＣＲ－ＳＳＣＰ方法检测常规石蜡包埋乳腺癌组织中ｐ５３基因点突变。结果显示６０例乳腺癌中，１４例有异常电泳带，表明这些病例相应ＤＮＡ片段中发生了点突变，其中４例位于第５～６外显子，３例位于第７外显子，７例位于第８外显子。免疫组化法检测突变型ｐ５３蛋白的表达，１３例为阳性，其中１１例ＳＳＣＰ检测到异常电泳带，另２例突变可能发生在所检测的基因片段以外。而５例ＳＳＣＰ分析发现基因突变者，免疫染色却为阴性，主要可能由于石蜡包埋组织中ｐ５３抗原性减弱或消失所致，因此，ＳＳＣＰ法检测ｐ５３基因点突变更为准确、可靠，而且可初步将突变定位在基因某一片段。本研究结果表明非同位素ＰＣＲ－ＳＳＣＰ是一简便、快速、有效的检测基因点突变的方法，特别适用于对大量标本基因点突变的筛选；ｐ５３基因点突变与突变型ｐ５３蛋白免疫染色阳性呈相关关系，但并不完全吻合。     

免疫组化和PCR—SSCP检测p53基因异常的一致性探讨

目的：探讨免疫组化检测ｐ５３蛋白表达和ＰＣＲ－ＳＳＣＰ检测ｐ５３基因突变的一致性。方法：单克隆抗体ＤＯ－７检测８５例非小细胞肺癌（ＮＳＣＬＣ）的ｐ５３蛋白表达，ＰＣＲ－ＳＳＣＰ检测其中３１例腺癌的ｐ５３基因突变。结果：８５例ＮＳＣＬＣ中ｐ５３蛋白表达阳性率为６８％（５８／８５），３１例腺癌中ｐ５３蛋白表达阳性率为６１％（１９／３１），１４例（４６％）出现ｐ５３基因５～８外显子突变，ｐ５３蛋白免疫组化和ＰＣＲ－ＳＳＣＰ检测ｐ５３基因突变无显著相关（χ２＝０．１，Ｐ＝０．７６），其一致率为５２％。结论：ｐ５３蛋白表达并不能很好地反映ｐ５３基因突变。     

肺癌中MTS1/p16和p53基因产物的表达与细胞增殖的关系

目的：研究肺癌中ＭＴＳ１／ｐ１６和ｐ５３基因产物的表达与细胞增殖的关系。方法：应用Ｓ－Ｐ免疫组织化学方法研究６２例肺癌组织中ｐ１６蛋白和ｐ５３蛋白的表达情况，并进行增殖细胞核抗原（ＰＣＮＡ）检测，计算细胞增殖指数（ｐｒｏｌｉｆｅｒａｔｉｏｎｉｎｄｅｘ，ＰＩ）。结果：６２例肺癌组织中ｐ１６蛋白和ｐ５３蛋白阳性率分别为５８．１％和５９．７％。腺癌ｐ１６蛋白的阳性率明显高于小细胞癌（Ｐ＜０．０５）；淋巴结转移阳性组ｐ１６蛋白的表达显著低于阴性组（Ｐ＜０．０５）；ＰＩ分级为Ⅱ级的ｐ１６蛋白表达显著高于Ⅳ级（Ｐ＜０．０５）。不同组织类型肺癌中ｐ５３蛋白的表达未见明显差异，淋巴结转移阳性组ｐ５３蛋白的表达高于阴性组（Ｐ＜０．０１）；不同ＰＩ分级中ｐ５３蛋白的表达，Ⅳ级明显高于Ⅰ级（Ｐ＜０．０５）和Ⅱ级（Ｐ＜０．０５），Ⅲ级明显高于Ⅰ级（Ｐ＜０．０５）和Ⅱ级（Ｐ＜０．０１）。ｐ１６蛋白低表达和ｐ５３蛋白过表达之间未见明显相关。结论：ｐ１６蛋白低表达和ｐ５３蛋白过表达均有促进肺癌细胞增殖的作用，ｐ１６蛋白的表达与肺癌的细胞分化有关，ｐ５３蛋白过表达对肺癌细胞的转移起重要作用。抑癌基因ｐ５３对ＭＴＳ１／ｐ１６基因无明显调控作     

DukesC期大肠癌原发灶与淋巴结转移灶中p53基因突变的比较

目的探讨大肠癌转移与ｐ５３基因突变的关系。方法用ＰＣＲ－ＳＳＣＰ检测７３例ＤｕｋｅｓＣ期大肠癌及４０例相应的淋巴结转移灶ｐ５３基因第５至第９外显子的突变。结果７３例原发灶有２７例（３７％），４０例淋巴结转移灶中有１９例（４７．５％）检测到ｐ５３基因突变，总突变率为４４％。在原发灶和转移灶同时检测的４０例中，有２１例（５２．５％）两者均未见ｐ５３突变；另１２例（３０％）两者均可见完全相同的突变条带；余下的７例（１７．５％）两者ＰＣＲ－ＳＳＣＰ的结果不一致。这７例经ＤＮＡ直接测序确定了ｐ５３基因突变位点。其中５例原发灶未见ｐ５３突变，而转移灶ｐ５３有明确突变；７例中２例转移灶可见ｐ５３有二个外显子突变，而原发灶仅一个或无突变点。结论多数ｐ５３基因突变发生在转移之前并持续存在于大肠癌进展至转移的整个时期，大肠癌的转移可伴有ｐ５３基因的新突变，或转移对ｐ５３基因突变有一定选择，提示ｐ５３基因突变可能与大肠癌转移相关。     

大肠肿瘤中抑凋亡基因bcl－2和p53蛋白产物的表达和意义

用免疫组化方法观察４５例大肠腺瘤和６１例大肠腺癌中ｂｃ１－２和ｐ５３蛋白的表达。结果显示：正常大肠粘膜中ｂｃ１－２和ｐ５３均未见表达，而大肠腺瘤及大肠腺癌阳性率均较正常组织明显增加（Ｐ＜０．０１）．大肠腺瘤中ｂｃ１－２表达位于不典型增生较重区域。大肠腺瘤ｐ５３表达随腺瘤直径增加而增加，其中≥２０ｍｍ组阳性率显著高于＜１０ｍｍ组（ｐ＜０．０５）。ｐ５３蛋白阳性率也随不典型增生程度增加而增高。ｐ５３表达与大肠腺癌分化程度及Ｄｕｋｅｓ分期有关。大肠腺瘤中ｂｃ１－２和ｐ５３蛋白的表达呈负相关。结果表明，ｂｃ１－２蛋白表达对大肠腺瘤的增殖有一定意义，ｐ５３在大肠腺瘤癌变和大肠腺癌进展中起重要作用。     

c-erbB-2、p53、bcl-2和nm23-H1在肺癌中的表达

目的：探讨ｃｅｒｂＢ２、ｐ５３、ｂｃｌ２和ｎｍ２３Ｈ１基因在肺癌发生发展过程中的作用。方法：用免疫组化ＡＢＣ法对原发性肺癌组织中４种基因的表达和突变进行检测。结果：５８例肺癌中,３１例（５３４５％）ｐ５３过度表达,１８例（３１０３％）ｂｃｌ２过度表达。ｃｅｒｂＢ２与ｎｍ２３Ｈ１在１０例小细胞肺癌（ＳＣＬＣ）中未见表达。而在４８例非小细胞肺癌（ＮＳＣＬＣｓ）中两者过度表达率均为５０％。ｃｅｒｂＢ２与ｎｍ２３Ｈ１表达呈正相关（Ｐ＜００５）。腺癌ｎｍ２３Ｈ１的表达明显高于鳞癌（Ｐ＜００５）。ｐ５３、ｂｃｌ２蛋白表达在肺癌分化程度中呈负相关（Ｐ＜００５）。ｎｍ２３Ｈ１、ｐ５３和ｂｃｌ２的表达与患者的生存率有关（Ｐ＜００５）。结论：ｃｅｒｂＢ２、ｐ５３、ｂｃｌ２和ｎｍ２３Ｈ１基因蛋白产物的检测对肺癌患者的诊治和预后评估有积极意义。     

DMBA-TPA-MNNG诱发小鼠皮肤肿瘤中p53基因突变

目的探讨ｐ５３基因与化学致癌剂诱发小鼠皮肤肿瘤的关系。方法运用ＰＣＲＳＳＣＰ和直接ＤＮＡ测序法，检测３７例ＤＭＢＡＴＰＡＭＮＮＧ诱发的小鼠皮肤肿瘤［包括６例乳头状瘤，１５例高分化鳞状细胞癌（ＳＣＣＩ）和１６例中分化鳞状细胞癌（ＳＣＣⅡ）］中，ｐ５３基因第５～８外显子的序列改变。结果乳头状瘤均未见ｐ５３基因突变，而２５８％（８／３１例）ＳＣＣ测得突变，其中ＳＣＣⅠ和ＳＣＣⅡ突变率分别为２６６％（４／１５例）和２５％（４／１６例）。在这８例肿瘤中共有９个突变，其中７个位于第８外显子，有７个为Ｇ→Ａ转换。结论ｐ５３基因突变发生于该三步化学致癌法诱发的小鼠皮肤乳头状瘤向ＳＣＣ恶性转化过程中。     

p53基因与溃疡性结肠炎

野生型ｐ５３蛋白具有抗细胞增殖功能，它可使ＤＮＡ有损伤的细胞生长停止于Ｇ１／Ｇ０期。散发性结直肠癌ｐ５３基因中常见点突变发生，发生于溃疡性结肠炎基础上的结直肠癌（ＵＣＡＣＲＣ）亦有很高的ｐ５３基因突变率。另一等位基因的杂合性缺失（ｐ５３ＬＯＨ）的发生也增高，加强溃疡性结肠炎病人ｐ５３基因点突变和ｐ５３ＬＯＨ的研究，可为监测ＵＣＡＣＲＣ的形成提供新方法。     

p53 expression in colorectal adenomas.

To assess the expression of p53 in premalignant lesions, we examined by immunohistochemistry benign colorectal adenomas (n = 72, measuring more than 6 mm and less than 95 mm in diameter) from patients without (group I, n = 23) or with (group II, n = 49) concurrent sporadic colorectal carcinomas. Using a panel of three monoclonal antibodies (PAb 240, PAb 421, PAb 1801) and two polyclonal antibodies (CM1, C19) immunohistological staining was demonstrated in 26% of the cases (19 of 72 adenomas, 7 of 23 from group I and 12 of 49 from group II). In the majority of the cases, p53 positive foci in the adenomas occurred in the most dysplastic areas, although focal positivity was detected in glands that were histologically normal. Expression of p53 protein was also detected in 21 of 30 (70%) colorectal carcinomas of group II. In two cases focal positive staining was observed in the polyps but not in the concurrent carcinomas. Non-neoplastic colonic mucosa and stromal lymphoid cells were negative in all cases examined. Over-expression of p53 in neoplastic tissues detected by immunocytochemistry is generally believed to correlate with the presence of mutation in the gene. This may not be an absolute rule, because in some hemopoietic malignancies, there is evidence that p53 protein may be detectable in the absence of an underlying mutation. These findings therefore represent the highest incidence in colorectal adenomas of abnormalities in the p53 protein expression, probably largely due to underlying mutations. This study also suggests that immunocytochemical demonstration of p53 protein may be a suitable method for the routine detection of subpopulations of cells which, by clonal expansion, could acquire a growth advantage within an adenoma during the neoplastic process.     

Absence of p53 gene mutation and infrequent overexpression of p53 protein in hepatoblastoma

Ten cases of hepatoblastoma were studies for overexpression of p53 protein by immunohistochemistry and for possible p53 gene mutation by single strand conformation polymorphism (SSCP) analysis and direct DNA sequencing of the polymerase chain reaction products. Only one case of the macrotrabecular type at stage IV showed overexpression of p53 protein. No DNA mobility shift was found in any of the cases studies by SSCP analysis. DNA sequencing performed on the case showing overexpression of p53 protein revealed no mutation within exons 5 to 8. The associated adrenal cortical carcinoma of the same case also showed overexpression of p53 protein, but no mutation of the p53 gene. These results indicate that mutation of the p53 gene is infrequent in hepatoblastoma. This observation supports the view that mutation of the p53 gene is not as important in the oncogenesis of childhood neoplasms as in adult cancers.     

骨肉瘤患者p53基因的改变及其与预后的关系

目的：研究人骨肉瘤中ｐ５３蛋白表达及Ｐ５３基因突变及其与预后的关系。方法：免疫组织化学染色观察人骨肉瘤中Ｐ５３蛋白表达，银染ＰＣＲ－ＳＳＣ民埋从瘤中Ｐ５３基因５－８外显子点突变状态。预后分析用Ｋａｐｌａｎ－Ｍｅｉｅｒ生存曲线及Ｌｏｇｒａｎｋ时序检验。结果：人骨肉瘤中Ｐ５３蛋白阳性率为２９．１７％，ＳＳＣＰ检测Ｐ５３基因５－８外显子占突变率为２３．９６％，Ｐ５３蛋主ＳＳＣ生患者与其阴性患者间总和生存     

Elevated content of p53 protein in the absence of p53 gene mutations as a possible prognostic marker for human renal cell tumors

P53 tumour suppressor gene expression was estimated immunohistochemically using DO-1 monoclonal antibody (recognising both wild-type and mutant p53) in 88 human renal tumours. Single strand conformation polymorphism (SSCP) analysis of possible mutations within exons 4–8 of the p53 gene was performed in 29 of the tumours (mostly immunostaining-positive cases). Obviously elevated p53 content was detected with DO-1 antibody in chromophobic cell carcinomas and most clear/chromophilic cell tumours (in chromophilic cell populations). In contrast, clear cell carcinomas demonstrated either complete absence of p53 expression or the presence of single immunopositive nuclei. Oncocytomas were completely negative. Additional immunostaining of the positive samples with mutant p53-specific Pab240 monoclonal antibody failed to detect immunopositive material. No p53 mutation was found in any of the samples analysed by SSCP. Our results suggest that the elevated p53 content in human renal cell carcinomas does not result from gene mutation and that p53 gene alterations are probably not an important mechanism in the development of human renal cell carcinomas. Accumulation of the wild-type p53 protein may be a useful prognostic marker indicating neoplastic progression and malignancy.     

大肠癌组织p14ARF与p53基因变异研究

目的：观察大肠癌组织p14ARF基因遗 传和表观遗传变化及p53基因突变状况，探讨两者改变的关系及p14ARF-p53通路功能 破坏在大肠癌发生中的作用。方法：应用PCR、直接测序、甲基化特异PC R和RT-PCR分别检测56例原发性大肠癌及相应癌旁正常组织p14ARF基因纯合性缺失、 突变、5′CpG岛甲基化、mRNA表达及p53基因突变状况。结果：①大肠 癌组织p14ARF总变异率为27% (15/56)，其中1例纯合性缺失，14例5′CpG岛甲基化,未见突变发生。②15例p14ARF基因变异的大肠癌组织其相应mRNA表达阴性（13例）或 低水平表达（2例），41例未发生基因变异的大肠癌组织和所有癌旁正常组织p14ARF mRNA均显示明显表达。③大肠癌组织p53突变率为48% (27/56)。④56例大肠癌组织中，12例 仅发生p14ARF变异，24例仅显示p53突变，3例同时有p14ARF 5′CpG岛甲基化 和p53突变，17例p14ARF和p53均无变异，p14ARF-p53通路总变异率为70%（39/ 56），p14ARF 5′CpG岛高甲基化与野生型p53状态有关（P＜0.05）。⑤低分化 腺癌组p14ARF变异率（44%，7/16）明显高于高中分化腺癌组（20%，8/40）（P＜ 0.05），但两组间p53突变率无显著差异（P＞0.05）。结论:①大肠癌p14ARF基因5′ CpG岛高甲基化是其表达失活的主要机制；②大肠癌p14 ARF基因高甲基化与p53基因突变可能是相互独立的事件，两者的失活可能各自出现于不同 的大肠癌亚组中；③p14ARF高甲基化或p53突变引起的p14ARF-p53通路功能 破坏在大肠癌的发生中具有重要作用。     

p53 protein expression in non-Hodgkin's lymphomas is infrequently related to p53 gene mutations

Accumulation of the p53 protein has been found In several types of lymphomas. However, p53 gene mutations have been infrequently demonstrated in some specific types of lymphomas. In the present study, a correlation between p53 immunoreactivity and p53 gene mutations in a large panel of non-Hodgkin's lymphoma (NHL) cases is attempted. A panel of 202 cases of NHL was evaluated by immuno-hlstochemical staining for p53 protein. All cases that were immunohistochemically positive for p53 protein were analyzed by the polymerase chain reaction (PCR) single strand conformation polymorphism (SSCP) method to identify mutations within the pS3 gene. In order to confirm the mutation, sequencing of PCR-amplified p53 gene segments was performed. Overexpression of p53 protein was found in 59 of the 202 cases of NHL, but only four of these 59 cases showed a shift on SSCP analysis, and point mutations were detected in three of them by the subsequent sequencing. p53 Immunoreactivity was generally greater in high-grade lymphoma. The results of this study suggest that Immunohistochemlcal reactivity for p53 protein is not a reliable Indicator of the presence of their structural alterations of p53 gene exons 4–9 in NHL.     

肺鳞状细胞癌中p53蛋白表达及p53基因突变的检测

目的 通过检测肺鳞状细胞癌及癌旁组织中Ｐ５３蛋白积聚及相同癌组织中Ｐ５３基因的突变，探讨Ｐ５３蛋白积聚及Ｐ５３基因突变在肺鳞癌发病中的意义。方法 采用免疫组化和银染－ＰＣＲ－ＳＳＣＰ方法检测１２０例肺鳞癌及癌旁肺组织中Ｐ５３蛋白的状况及相同鳞癌组织中，Ｐ５３基因５、６、７、８外，显子突变的情况。结果 Ｐ５３蛋白了性率为５２．５％，Ｐ５３基因突变率为５６．７％，突变便数在第５、６、７、８外显子的分布     

The relationship between the gene mutation of p53 and the protein expression of p53 and Ki-67 in non-Hodgkin's lymphomas

The relationship between the mutation of the p53 gene and the expression of the p53 protein and the Ki-67 antigen has been investigated in 115 cases with non-Hodgkin's lymphoma, using the immunohistochemical double staining technique, single-strand conformational polymorphism and DNA sequencing methods. Eighteen cases showed more than 10% of p53+ cells and the others showed a few p53+ cells presented sporadically. Alterations in the p53 gene were detected in six cases with B cell type, consisting of five cases with point mutation and one case with point mutation and 15 base pairs deletion. These six cases showed a high percentage of p53+ cells and five cases revealed that the percentage of p53+ cells was higher than that of Ki-67+ cells (p53+ cells > Ki-67+ cells). Excluding the six cases with mutation of the p53 gene, all cases revealed that the percentage of p53+ cells was less than that of Ki-67+ cells (p53+ cells < Ki-67+ cells). Moreover, there was a positive correlation between expression of the p53 protein and of the Ki-67 antigen in histologic types of B cell lymphomas and of T cell lymphomas, respectively, except in small non-cleaved (Burkitt's) and lymphoblastic types. Therefore, sporadic cases showing p53+ cells > Ki-67+ cells revealed alteration of the p53 gene, and expressed abnormal p53 protein (mutant form). Most cases showing p53+ cells < Ki-67+ cells expressed normal p53 protein (wild type), and may reflect the rapid proliferation rate.     

Expression of p53 protein in infiltrating and in-situ breast carcinomas.

Five antibodies directed against the whole or part of p53 protein have been used to detect the protein immunohistochemically in 70 infiltrating breast carcinomas and 10 ductal carcinomas in situ. Mutations are known to occur in different conserved domains, and the antibodies employed spanned the expected sites. p53 protein was identified in 53 per cent of infiltrating carcinomas using the antibodies PAb 240, PAb 1801, C19, and JG8. The antibody PAb 421 detected the protein in 31.5 per cent; all positive with the other antibodies. Well-differentiated oestrogen receptor-positive tumours had a low incidence of p53 detection. Variation in the percentage of reactivity was seen between carcinomas and in some cases between different antibodies in the same cancer. Those carcinomas with a high percentage of positive cells with all antibodies were more likely to have metastasized to nodes, be at an advanced stage, and be oestrogen receptor-negative/epidermal growth factor receptor-positive. There was no significant correlation with c-erbB-2 protein expression or retinoblastoma protein loss. p53 protein was detected in a high proportion of cells in three of the six comedo ductal carcinomas in situ studied but either not at all or at a lower level in tumours of the cribriform type. p53 mutations are common in breast carcinomas, but heterogeneity within individual tumours is frequent. Marked expression of p53 appears to relate to tumour progression.     

Ⅲ期鼻咽癌p53蛋白表达与生物学行为及预后的关系及其与p53基因突变、潜伏膜蛋白-1表达的相关研究

目的：探讨Ⅲ期鼻咽分化型非角化性鳞癌中P53蛋白的表达与生物学行为及预后的关系,以及P53蛋白表达与P53基因突变和潜伏膜蛋白（LMP）－1蛋白表达的关系。方法：应用免疫组织化学EnVison法和聚合酶链反应－单链构象多态性分析（PCR－SSCP）检测58Ⅲ例鼻咽分化型非角化性鳞癌标本。结果：58份Ⅲ期鼻咽分化型非角化性鳞癌中,P53蛋白总阳性率为65．5％（3858）,生存期在5年以下的患者阳性率为82．1％（32／39）,5年以上患者31．6％（6／19）,两组间P53蛋白表达差异有显著性意义（P＜0．05）,P53蛋白表达在有预底侵袭组为74．4％（29／39）,无颅底侵袭组为47．4％（919）,两组间差异有显著性意义（P＜0．05）,PCR－SSCP检测15例P53蛋白阳性的病例,P53基因5－8外显子突变率为0（0／15）,LMP－1蛋白免疫组织化学总阳性率为72．4％（42／58）,与P53蛋白表达呈正相关关系（r=0.504)。结论：本组Ⅲ期鼻咽分化型非角化性鳞癌患者中P53蛋白表达与P53基因突变无相关关系。与LMP－1蛋白的表达相关,P53蛋白的表达阳性率随着患者生存期的延长而降低,有颅底侵袭组P53蛋白表达阳性率高于无颅底侵袭组。