p53蛋白在卵巢癌组织中的表达及其意义

目的：探讨ｐ５３蛋白表达与ＤＮＡ倍体、临床分期、病理分级及预后的关系。方法采用流式细胞仪双参数检测了５４例临床卵巢肿瘤手术标本的ＤＮＡ含量和Ｐ５３蛋白表达。其中良性肿瘤６例,卵巢癌４８例。同时采用免疫组化方法检测上述样本２０例的石蜡切片。观察Ｐ５３蛋白表达。结果经统计分析,采用免疫组化ＡＢＣ法和流式细胞仪法检测的Ｐ５３蛋白表达者有明显一致性（Ｐ〈０．０１）。良恶性肿瘤组织中ｐ５３蛋白表达有显著性差     

人乳腺癌组织p53基因突变及过度表达的临床意义

应用聚合酶链反应－单链构象多态（ＰＣＲ－ＳＳＣＰ）分析和流式细胞术（ＦＣＭ），研究３０例患者乳膛癌组织Ｐ５３基因５￣８外显子的突变情况、Ｐ５３蛋白免疫组化检出情况、ＤＮＡ倍体及雌激素受体（ＥＲ）含量；并与各生物学参数进行了相关研究，以探讨Ｐ５３基因与乳腺癌发生、发展的关系。结果提示：１．全组１４例（４６．７％）发现Ｐ５３基因突变，其中１例为原位导管癌。２．Ｐ５３蛋白检出阳性率为５３．３％（１６／３     

肺癌组织抑癌基因P53异常分析

Ｐ５３基因５－８外显子ＰＣＲ产物的ＤＮＡ序列分析及石蜡包埋组织Ｐ５３鼠抗人单克隆抗体（Ｄｏ－１）免疫组化分析发现：５７例原发性肺癌中３７例有Ｐ５３蛋白积累。ＤＮＡ／ＰＣＲ产物直接测序发现：９／１２例小细胞肺癌（ＳＣＬＣ），８／１５非小细胞肺癌（ＮＳＣＬＣ）Ｐ５３基因突变，错义突变为１２／１７，Ｇ→Ｔ突变为６／１７。免疫组化发现：１１／１９例ＳＣＬＣ，１９／３７例ＮＳＣＬＣ Ｐ５３蛋白染色阳性。４／     

散发性结直肠癌中p53蛋白表达与微卫星不稳定性的关系

目的 探讨散发性结直肠癌中微卫星不稳定性（ＭＩＮ）发生与Ｐ５３蛋白表达的相关性及其意义。方法：采用微卫星ＤＮＡ－ＰＣＲ－银染色法检测６７例散发性结直肠癌４条染色体上６个微卫星位点的ＭＩＮ,应用ＳＡＢＣ方法检测Ｐ５３蛋白表达。结果 以２个或２个以上位点有ＭＩＮ定义为复制误差阳性（ＰＣＲ＋）,ＲＥＰ＋率４１．８％（２８．．６７）。５０例Ｐ５３蛋白表达阳性６０．６７,７４．６％）。ＲＥＰ＋组Ｐ５３蛋白阳     

胃癌P53基因突变和蛋白表达的研究

为了解中国人胃癌Ｐ５３基因突变情况，应用银染聚合酶链反应－单链构象多态（ＰＣＲ－ＳＳＣＰ）技术对Ｐ５３基因第５、６、７、８外显子进行检测。对来自癌组织ＤＮＡ和癌旁正常组织ＤＮＡ的ＰＣＲ－ＳＳＣＰ电泳带迁移作对比分析，发现３７例胃癌中，１１例胃癌样品电泳带迁移异常，依据ＤＮＡ单链构象与分子电泳迁移的关系，研究结果表明：胃癌Ｐ５３基因５－８外显子的突变率为３０％。微波处理ＡＢＣ法检测突变型Ｐ５３蛋白表     

p53功能在肝癌中表达的意义

为探讨Ｐ５３功能及某些生物学行为肝癌中表达的意义。方法：收集１９９７年１月至１９９８年１２月５２例原发性肝癌的临床资料，并对这５２例手术切除的肝癌标本，分别用Ｗｅｓｔｅｒｎ ｂｌｏｔ结合ＤＮＡ损伤诱导的方法检测野生型Ｐ５３的功能，免疫组化检测Ｐ５３蛋白的表达，ＰＣＲ／ＳＳＣＰ方法检测Ｐ５３基因在第５－８外显子的丢失情况及免疫组化检测肿瘤增殖细胞核抗原（ＰＣＮＡ）指数。结果：５２例标本中，３１例（５     

胆管癌p53基因突变和蛋白表达的研究

目的了解胆管癌ｐ５３基因突变和蛋白表达的变化。方法应用聚合酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）银染技术检测胆管癌中ｐ５３基因突变，并用亲和素－生物素－过氧化物酶复合物（ＡＢＣ）免疫组织化学染色方法检测胆管癌ｐ５３蛋白的表达。结果１１例胆管癌中，３例胆管癌样品ＰＣＲ－ＳＳＣＰ单链电泳带迁移异常，依据ＤＮＡ单链构象与分子电泳迁移的关系，胆管癌ｐ５３基因５～８外显子的突变率为２７％；６例有ｐ５３蛋白异常表达，阳性率为５５％。结论导致ｐ５３蛋白异常表达的ｐ５３基因突变是胆管癌的一种分子结构改变，可能在胆管癌的发生和发展中起着一定的作用。     

p53和c—myc异常表达与膀胱癌多药耐药性

为探讨ｐ５３和ｃ－ｍｙｃ基因异常表达与膀胱癌多药耐药性（ＭＤＲ）的关系，应用ＡＢＣ免疫组织化学方法检测６７例膀胱癌癌细胞中ｐ５３和ｃ－ｍｙｃ的表达产物及多药耐药基因（ｍｄｒ－１）产物Ｐ－ｇｐ。结果显示ｐ５３突变蛋白蓄积的膀胱癌中Ｐ－ｇｐ阳性表达率９２．３％；ｐ５３阴性者Ｐ－ｇｐ阳性率为３６．６％。Ｐ－ｇｐ表达与ｐ５３突变蛋白蓄积呈显著正相关（ｒ＝０．６４，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，使膀胱癌细胞获得ＭＤＲ表型     

银染PCR—SSCP方法检测恶性纤维组织细胞瘤P53基因点突变

应用银染ＰＣＲ－ＳＳＣＰ方法检测常规石蜡包埋恶性纤维组织细胞瘤（ＭＦＨ）中ｐ５３基因点突变。结果显示，１６例ＭＦＨ中，９例ＳＳＣＰ阳性，表明这些病例相应ＤＮＡ片段中发生了点突变，其中第５、６、７、８外显子ＳＳＣＰ阳性出现的例次数分别为１、４、４、３次（２例转移的病例同时有６、７、８三个外显子异常）。微波处理ＡＢＣ法检测突变型ｐ５３蛋白表达，１０例为阳性，ＳＳＣＰ与ｐ５３蛋白表达的阳性符合率为９０．０％，ｐ５３基因点突变及蛋白表达与ＭＦＨ的组织学亚型无关。ｐ５３基因突变可能在ＭＦＨ的发生发展及转移中起作用。本研究结果表明，银染ＰＣＲ－ＳＳＣＰ是一简便、快速、有效的检测基因点突变的方法。     

胃癌P53基因表达与肿瘤细胞增殖活性的研究

目的：研究胃癌Ｐ５３基因表达以及与ＤＮＡ含量和细胞增殖活性的关系。方法：采用流式细胞术和细胞免疫荧光肝癌Ｐ５３蛋白、ＤＮＡ、和细胞周期。结果：５６例胃癌Ｐ５３蛋白阳性３２例（５７．１２％），正常胃粘膜和良性病变组均为阴性。Ｐ５３蛋白阳性者多为异倍体肿瘤，ＤＩ值、ＳＰＦ和ＰＩ值均明显高于阴性才，两组差异显著。结论：Ｐ５３基因突变可能是促进胃癌发生发展和导致肿瘤细胞增异常活跃的重要因素。     

新鲜肿瘤标本活细胞与固定细胞的 DNA含量分析比较

目的：建立一种以新鲜细胞进行临床肿瘤DNA含量分析的方法。方法：血液肿瘤取开肝素抗凝外固血或骨髓样本，用淋巴细胞分层液分离纯 化单个核细胞；实体瘤组织则用机械法制成单细胞悬液。将细胞分成两份，一份酒精固定过夜，流式细胞术常规方法测定DNA含量，另一份直接用PIPES（piperazine-N,N‘-bis[2-ethanesulfonic acid]disodium salt)缓冲液配制的碘化丙锭（propidium iodide,PI)染色，以流式细胞术进行DNA含量测定，并与同期固定细胞的DNA含量检测结果进行比较。结果；同类型的肿瘤细胞，其固定前后G0／G1期细胞峰的变异系数（ocefficient of variation,CV)没有明显差异（t检验，P＞0.05）。重复实验结果显示，新鲜细胞DNA含量分析结果稳定。结论：用PIPES缓冲液配制的PI染液染色新鲜细胞，可以对临床肿瘤的细胞增殖状况和DNA倍性进行分析。     

Nuclear DNA content-derived parameters correlated with heterogeneous expression of p53 and bcl-2 proteins in clear cell renal carcinomas

BACKGROUND: p53 and bcl-2 are two key genes involved in cell cycle and cell death regulation. Altered expression or mutation of these genes has been found in human cancers and also has been identified in clear cell renal carcinoma (RCC). Their role in RCC progression, however, is still unclear. By contrast, the prognostic significance of ploidy and S-phase fraction (SPF) have been studied extensively in RCC. To better characterize the biologic role of p53 and bcl-2 oncoproteins in RCC, we offer a multisample correlative analysis of the expression of these two proteins with ploidy and SPF. METHODS: Ploidy and SPF along with p53 and bcl-2 expression were analyzed in 296 specimens, selected by multiple sampling of 33 consecutive operable RCCs. The expression of p53 and bcl-2 proteins was studied by immunohistochemistry, and SPF and tumor ploidy were studied by flow cytometry. RESULTS: In our study, 4 of 32 (12.5%) were found to be diploid, and 28 of 32 (87.5%) cases showed an abnormal DNA content. Among the aneuploid tumors, 14 of 28 (50%) were multiploid. Heterogeneous DNA content was detected in 21 of 32 (65.6%) tumors and was correlated with the more advanced Robson stage tumor (P = 0. 03). Intratumor heterogeneity also was detected for p53 and bcl-2 protein expression. Expression of p53 protein correlated with the lack of bcl-2 protein expression (P = 0.0032), aneuploidy (P < 0. 0001), and high SPF (P = 0.0006), whereas bcl-2 expression was associated with a normal DNA content (P < 0.0001) and low SPF (P = 0. 035). CONCLUSIONS: Within each RCC, p53 and bcl-2 expression is markedly heterogeneous. Our results depicted a scenario in which bcl-2 protein, expressed by normal renal parenchyma, is still present in euploid cell clones of RCC but disappears during the progression of renal neoplasm toward a more aggressive phenotype characterized by overexpression of p53 protein, aneuploidy, and high SPF.     

乳腺癌P53基因蛋白的表达与临床预后的关系

应用流式细胞术对４０例乳腺癌细胞Ｐ５３蛋白的表达进行了定量研究，并探讨其与病理组织学分级，ＤＮＡ倍体和临床预后的关系。结果表明，Ｐ５３表达与乳腺癌组织学分级有关。ＤＮＡ异倍体乳腺癌Ｐ５３表达量（ＦＩ值）明显高于二倍体乳腺癌。Ｐ５３阳性表达的乳腺癌，其生存期（４．７±２．９年）和五年生存率（３４．６％）明显低于Ｐ５３阴性表达的乳腺癌。Ｐ５３基因的突变，可能是一个重要的、新的预后指标。     

滤泡性淋巴瘤与反应性淋巴滤泡增生的免疫组化及DNA图像分析研究

目的：观察滤泡性淋巴瘤（ＦＬ）与反应性淋巴滤泡增生（ＲＦＨ）中滤泡生发中心细胞（ＧＣＣｓ）核ＤＮＡ含量、倍体情况以及ｂｃｌ－２癌基因蛋白、免球蛋白轻链（ＩｇＬ）表达情况，探讨它们对于二都鉴别诊断的意义。方法：对２１例ＦＬ及２１例ＲＦＨ进行ｂｃｌ－２蛋白、Ｋｆｐｐａ、Ｌａｍｂｄａ轻链蛋白免疫组化检测及ＤＮＡ图像细胞分析。结果：６１．９％ＦＬ中ＧＣＣｓ有ｂｃｌ－２表达而ＲＦＨ的ＧＣＣｓ均为ｂｃｌ－２阴     

Lack of diagnostic-value of DNA content and p53 immunostaining in normal, hyperplastic and neoplastic parathyroid tissue

The aim of this project was to study the diagnostic value of DNA content and p53 protein expression in normal, hyperplastic and neoplastic parathyroid lesions. Tissue samples of 74 parathyroid glands from 34 patients with primary hyperparathyroidism were studied by DNA flow cytometry and p53 immunostaining. In 9 of 23 patients (39%) with parathyroid adenoma, a nondiploid cell population was present. Some normal looking glands removed from the same patients also had a nondiploid DNA index. Multiglandular hyperplasia was found in 11 patients, and in 5 of these (45%) the histograms showed nondiploid cells. The proliferative activity was generally low and S-phase fraction did not differ in glands with hyperplasia or adenoma, when compared with normal looking glands. One single case of hyperplasia showed a weak p53 positivity in scattered nuclei, probably representing wild type p53 protein. Thus, our present results suggest that DNA content and p53 protein staining are of no value in the routine work up of parathyroid glands removed from patients with primary hyperparathyroidism.     

卵巢粘液性肿瘤P53蛋白的表达及与DNA·论著·含量关系的研究

目的研究卵巢粘液性肿瘤中Ｐ５３蛋白的表达及与病理组织学分型,分级,临床分期,ＤＮＡ含量的关系。方法应用免疫组织化学及图像分析技术对６５例各种组织学类型卵巢粘液性肿瘤中Ｐ５３蛋白的表达及ＤＮＡ倍体进行了观察。结果Ｐ５３蛋白表达率在囊腺瘤为阴性,交界性囊腺瘤为１７．４％,囊腺癌为５０．０％,囊腺癌的表达率显著高于囊腺瘤（Ｐ＜０．００１）和交界瘤（Ｐ＜０．０２５）。Ｐ５３蛋白的表达与囊腺癌不同组织学分级和临床分期之间无相关性（Ｐ＞０．０５）。图像定量分析显示,囊腺癌Ｐ５３蛋白阳性细胞平均积分光密度值（３．６３±２．２０）显著高于交界瘤的值（１．６８±３．４９）,（Ｐ＜０．００１）。ＤＮＡ含量图像分析显示,Ｐ５３蛋白阳性组,肿瘤细胞ＤＮＡ指数总体平均值（１．８８±０．２３）显著大于Ｐ５３蛋白阴性组的ＤＮＡ指数的总体平均值（１．３４±０．３３）,（Ｐ＜０．００１）;≥５ｃ的肿瘤细胞百分率（３４．１％）显著高于阴性组（６．９％）（Ｐ＜０．００５）。结论Ｐ５３基因突变参与了卵巢粘液性肿瘤由良性向恶性转化的过程,Ｐ５３蛋白的表达可能作为该肿瘤恶性转化的标志,Ｐ５３蛋白表达与ＤＮＡ含量关系密切。     

p53蛋白在低剂量电离辐射诱导EL-4细胞适应性反应中的变化

目的：研究低剂量电离辐射诱导EL-4细胞适应性反应中p53 mRNA及蛋白的表达,为探讨低剂量电离辐射诱导细胞适应性反应的DNA损伤修复机制提供实验依据。方法：将EL-4细胞分为空白对照组,单纯照射组（其照射剂量分别为1 Gy、2 Gy和3 Gy）,以及诱导适应性反应照射组（其照射剂量分别为75 mGy＋1 Gy、75 mGy＋2 Gy、75 mGy＋3 Gy）。应用流式细胞仪和RT-PCR方法分别检测p53蛋白及mRNA水平表达。结果：单纯照射组p53蛋白水平较对照组显著升高（P〈0.01）,预先给予75 mGy照射诱导的适应性反应组其p53蛋白水平较单纯照射组则显著降低（P〈0.01）。p53 mRNA水平表现出与p53蛋白表达相同的变化。结论：p53蛋白在诱导适应性反应照射组中低表达,说明其在低剂量电离辐射诱导适应性反应中可能起重要作用。     

Expression of p53 protein and DNA flow cytometry in gastric adenocarcinoma: implications in patients treated with adjuvant etoposide, adriamycin and cisplatin

AIMS AND BACKGROUND: We evaluated the prognostic value of p53 protein, DNA content and S-phase fraction in patients with adenocarcinoma of the stomach or the gastroesophageal junction treated with adjuvant etoposide, doxorubicin and cisplatin. METHODS AND STUDY DESIGN: Thirty-five consecutive patients with stage II or III gastric or gastroesophageal junction adenocarcinoma treated with at least two cycles of adjuvant etoposide, doxorubicin and cisplatin after curative gastric resection were included. The expression of p53 protein was determined by immunohistochemistry and DNA content by flow cytometry. The presence of p53 expression and DNA content was compared with clinicopathological features. RESULTS: Median age was 54 years (range, 31-71). P53 expression was detected in 42.9% (15 of 35) of gastric cancer tissues of the patients. Aneuploidy was observed in 31.4% of patients, and S-phase fraction was more than 10% in 22.9%. P53 immunoreactivity (33.3% vs 47.8%) was more common in advanced disease. There was no association among p53 immunoreactivity, DNA content and S-phase fraction. We also found no significant relationship between p53 immunoreactivity, DNA content, S-phase fraction or other clinicopathological parameters. In univariate analysis, the involvement of lymph nodes was a significant predictor of a poor outcome (P = 0.001). Also, p53-positive patients had a poor survival close to the level of significance (P = 0.051). Likewise, p53 immunoreactivity (P = 0.0071), in addition to lymph node involvement (P = 0.0016), were the independent prognostic factors in multivariate analysis. CONCLUSIONS: This trial supports the results of previous reports that p53 immunoreactivity is a prognostic factor for patients with adenocarcinoma of stomach or gastroesophageal junction treated with adjuvant chemotherapy.     

RNA干扰p53对肝细胞癌SK-Hep-1细胞细胞周期的影响

背景与目的: 肝细胞癌是严重威胁人类健康的恶性肿瘤之一, p53又是重要的抑癌基因,而RNA干扰(RNAinterference, RNAi) 不仅是一种经济、快捷、高效的抑制基因表达的技术手段, 而且有可能在肿瘤基因治疗方面提供新的途径。因此, 我们将外源性p53双链小RNA(smalldoublestrandedRNA, dsRNA) 瞬时转染肝癌SK- Hep -1细胞系, 观察其对肝癌细胞P53和P21蛋白表达及细胞周期的影响。方法: 合成的p53dsRNA和EGFPdsRNA, 分别以200ng和400ng的p53dsRNA用脂质体转染法转染SK- Hep- 1细胞, 同时设0. 9%NaCl空白对照和EG- FP及EGFP+EGFPdsRNA阳性对照组, 每组3个复孔, 分别在转染24h和48h时用流式细胞仪进行细胞周期检测, 对转染p53dsRNA48h后应用WesternBlot检测P53和P21蛋白的表达,初步探讨p53的dsRNA干扰对肝癌细胞影响的机制。结果: SK- Hep -1细胞在转染200ngp53dsRNA后G0 -G1 期细胞明显减少, 24h时与空白对照相比减少了52 .53%, 与转染同剂量EG FP+EGFPdsRNA的阳性对照组相比也减少了50 .29% (P<0. 05); S期细胞明显增多, 24h时与空白对照相比增加了146 8%, 与转染同剂量EGFP+EGFPdsRNA的阳性对照组相比增加了128. 62% (P<0 .05); G2-M期细胞也明显增多, 24h时与空白对照相比增加了30. 56% (P<0. 05),