Molecular detection of genetic defects in congenital adrenal hyperplasia due to 21-hydroxylase deficiency: A study of 27 families

Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase (21-OHase) deficiency is inherited as an autosomal recessive trait. Patients can present with the salt wasting, simple virilizing or a non-classical form of the disease. The gene for P450C21, the enzyme carrying 21-OHase activity, has been mapped to the major histocompatibility complex on chromosome 6p. Using molecular hybridisation techniques we have studied the genetic defect in 27 families with one or more affected off-spring diagnosed and treated at the University Hospital of Essen. DNA samples were digested with restriction endonucleaseTaqI,PvuII,BglII, andEcoRI and analysed by Southern blot hybridisation with the cDNA probe pC21/3c. Eleven of 40 haplotypes associated with the salt wasting form were found to have a large deletion of 30 kb affecting the 5 end of the active 21-OHase gene and the 3 end of the closely linked pseudogene. Results in another 11 cases are compatible with gene conversion; 18 cases were not informative. The 30 kb deletion was associated with a combination of the HLA antigens Bw47 and DR7 in 7 of 11 cases. In the haplotypes with gene conversion, no linkage disequilibrium to HLA antigens was found. No apparent gene alterations were detected in simple virilizing and non-classical haplotypes. The direct detection of the genetic defect in 55% of the salt wasting haplotypes may help to improve predictive testing in families with CAH.     

葡萄胎p53、p21~(CIP1)及p185蛋白表达与恶变的关系

目的 :探讨葡萄胎p5 3、p2 1CIP1及p185蛋白表达与恶变的关系及其临床特点。方法 :免疫组化法检测葡萄胎标本中p5 3、p2 1CIP1及p185蛋白的表达 ,以侵蚀性葡萄胎及绒癌为对照 ,并回顾性分析其临床资料。结果 :葡萄胎组p5 3、p2 1CIP1、p185蛋白阳性表达率分别为 35 % (35 / 10 0 ) ,71% (71/ 10 0 )及 36 % (36 / 10 0 ) ,与恶性对照组相比均有显著性差异 (P <0 .0 5 ) ;而完全性与部分性葡萄胎 ,完全性葡萄胎恶变组与非恶变组之间差异均无显著性 (P >0 .0 5 ) ,但恶变组p2 1CIP1表达有降低趋势。结论 :p5 3、p2 1CIP1及p185蛋白表达改变与葡萄胎恶变无确定关系 ,但可作为晚期现象出现于恶性滋养细胞肿瘤中 ,其中p2 1CIP1蛋白表达降低提示滋养细胞有向恶性转化的倾向     

Utility of peptide–protein affinity complexes in proteomics: identification of interaction partners of a tumor suppressor peptide*

Abstract: We used a N‐biotinylated peptide analog of the C‐terminal domain of the tumor suppressor protein, p21^cip1/waf1 to elucidate peptide/protein interacting partners. The C‐terminal domain of p21^cip1/waf1 protein spanning 141–160 amino acid residues is known to bind PCNA and this interaction is important in many biological processes including cell‐cycle control. This C‐terminal 20‐mer efficiently extracts PCNA in the presence of a variety of N‐ or C‐terminally attached affinity tags. Using difference silver stained 2D gels combined with in‐gel tryptic digests, we identified the difference spots using MALDI‐TOF mass spectrometry‐based peptide mass fingerprinting followed by a database search using profound against NCBIs human nonredundant protein sequence data bank. Identified spots include the p48 subunit of chromatin assembly factor‐1, the heat shock 70 protein analog BiP, calmodulin, nucleolin and a spot similar in size to dimeric PCNA. In contrast, microcapillary ion‐trap LC‐MS/MS analysis of a tryptic digest of entire affinity extracts derived from both control and experimental runs followed by database searches using sequest confirmed the presence of most of the above proteins. This strategy also identified hnRNPA1, HPSP90α, HSP40 and T‐complex protein 1, a protein similar to prothymosin, and a possible allelic variant of the p21^cip1/waf1 protein. The use of N‐biotinylated peptide derived from the C‐terminal domain of p21^cip1/waf1 protein in proteomic analysis exemplified here suggests that peptides obtained from intracellular functional screens could also potentially serve as efficient baits to discover new drug targets.     

乳腺癌ER、c-erbB2、P21、PCNA表达及其意义

[目的]探讨ER、c-erbB2、p21、PCNA在乳腺癌的表达及其临床意义.[方法]应用免疫组化SP法,检测300例乳腺癌ER、c-erbB2、p21、PCNA表达.[结果]ER、c-erbB2、p21、PCNA在乳腺癌的阳性表达率分别为57.5%、47.3%、56%、663%;c-erbB2、p21、PCNA的表达随着组织分级升高而增强,ER的表达随着组织分级升高而降低;ER表达与生存期亦未见明确的关系,而c-erbB2、p21、PCNA阳性表达率与生存期呈负相关关系.[结论]ER、c-erbB2、p21、PCNA是判断乳腺癌分化程度、预后有价值的指标.     

非小细胞肺癌患者血清CEA及CYFRA21-1水平动态分析

目的:探讨治疗前后不同时期非小细胞肺癌患者血清CEA、CYFRA21蛳1水平的变化规律及临床意义。方法:分别于化疗前、后采集非小细胞肺癌患者血清样本,采用放射免疫分析法检测患者血清CEA,CYFRA21蛳1含量,并对检测结果进行比较分析。结果:非小细胞肺癌患者CEA阳性率为59.2 %,其中鳞癌47.6 %,腺癌72.9 %。CYFRA21蛳1阳性率为65.1 %,其中鳞癌71.9 %,腺癌57.1 %。二者均随临床TNM分期增加而升高,手术后及化疗有效者CEA、CYFRA21蛳1逐渐降低。结论:动态检测患者CEA,CYFRA21蛳1水平对非小细胞肺癌的诊断、病程及转移情况、疗效及预后判断、监测复发有重要意义。     

卵巢癌细胞DNA倍体、p21及p53基因表达产物定量分析

目的　探讨卵巢癌DNA倍体、p2 1及 p5 3基因定量表达情况及其临床意义。 方法　应用流式细胞术 (FCM )和荧光免疫技术 ,对 3 9例卵巢癌组织的卵巢癌细胞和 9例正常卵巢组织的卵巢细胞的染色体倍体、p2 1基因和 p5 3基因表达产物进行定量分析。结果　在卵巢癌组织中DNA的异倍体率、p2 1、p5 3基因蛋白产物的阳性表达率均明显高于对照组 ,两者比较均有非常显著性差异 (P <0 .0 1)。结论　应用FCM定量测定卵巢癌细胞中的DNA倍体、p2 1及 p5 3基因表达产物 ,在临床上具有重要的参考价值     

P21WAF1, P27KIP1, TP53 and C-MYC analysis in 204 ovarian carcinomas treated with platinum-based regimens.

BACKGROUND: The prognostic and predictive value of cell cycle regulatory proteins in ovarian cancer has not been established. We evaluated the clinical and biological significance of P21(WAF1), P27(KIP1), C-MYC, TP53 and Ki67 expressions in ovarian cancer patients. MATERIALS AND METHODS: Immunohistochemical analysis was performed on 204 ovarian carcinomas of International Federation of Gynecology and Obstetrics (FIGO) stage IIB to IV treated with platinum-based chemotherapy. Multivariate analysis with Cox and logistic regression models was performed in the whole group, and in the TP53-negative and TP53-positive subgroups. RESULTS: High P21(WAF1) labeling index (LI) was an independent positive predictor of platinum-sensitive response (P = 0.02). Overall survival was positively influenced by P21(WAF1) LI (P = 0.02) or by P21(WAF1) plus P27(KIP1) LI (P = 0.004) in the TP53-negative group only. Ki67 LI showed borderline association with disease-free survival (P = 0.05). Growth fraction was negatively associated with P21(WAF1) and P27(KIP1) indices in the TP53-negative group (P = 0.023 and 0.008, respectively), and these associations were borderline or lost in the TP53-positive group. Endometrioid and clear cell carcinomas differed from other carcinomas by having a low incidence of TP53 accumulation, a high incidence of C-MYC overexpression (70%) and a low median Ki67 LI (all with P <0.001). CONCLUSIONS: We have shown an independent predictive value of P21(WAF1) LI in ovarian carcinoma patients. The prognostic value of P21(WAF1) and P21(WAF1) plus P27(KIP1) LI was determined by TP53 status. A high frequency of C-MYC overexpression in endometrioid and clear cell carcinomas may suggest its role in the development of these tumor types.     

VEGF、p21ras在鼻咽癌组织中的表达及其临床意义的研究

目的 :研究 VEGF及 p2 1ras蛋白表达与鼻咽癌临床生物学特性及预后的关系 ,更好地评估预后 ,指导临床治疗。方法 :用免疫组化法测定 81例鼻咽低分化鳞癌 VEGF及 p2 1ras蛋白表达 ,对阳性和阴性病例进行统计分析。结果 :VEGF在不同分期、颈淋巴结转移、放疗后远处转移病例中的表达有显著性差异 (P<0 .0 5 ) ,p2 1ras蛋白高表达 ,但与不同分期、局部复发、颈淋巴结转移、远处转移无关。单因素分析显示影响鼻咽癌生存的预后因素有临床分期 (P<0 .0 5 )和癌组织中 VEGF表达水平 (P<0 .0 5 )。 Cox逐步回归模型分析显示临床分期是鼻咽癌的独立预后因素。结论 :鼻咽癌组织中 VEGF表达与临床 N分期、颈淋巴结转移、远处转移有显著相关 ;单因素分析 VEGF表达与鼻咽癌的预后呈正相关 ,可考虑作为预测鼻咽癌患者放射治疗后远处转移和预后的因素之一。 p2 1ras蛋白高表达 ,但与颈淋巴结转移、远处转移、局部复发无关 ,可作为联合检测的指标之一用于肿瘤的早期诊断。 VEGF表达与 p2 1ras蛋白表达无相关性。     

Haptoglobin-related protein as a serum marker in malignant lymphoma

A novel serum 21 kDa haptoglobin-related protein (Hpr) was investigated in patients with malignant lymphoma, to evaluate its correlation with clinical and histologic features at presentation and its possible role as a tumor marker for patient outcome. One hundred fifty eight serum samples were taken from 88 patients with non-Hodgkin’s lymphoma (n=58) and Hodgkin’s disease (n=30) at presentation and in the course of follow-up. Sera from 61 healthy volunteers served as normal controls. Serum Hpr levels in the lymphoma patients (median 430xl0³ u/ml, range 0-4000xl0³) were significantly higher than in the control group (median 68xl0³ u/ml, range 0-180xl0³) (p=0.0001). Higher median Hpr values were detected in patients with advanced disease (p=0.013), “B” symptoms (p=0.029) and in males (p=0.053). There was also a significant correlation between Hpr and erythrocyte sedimentation rate (p=0.028). Serial determinations showed a significant decrease of the initial Hpr values obtained after treatment in 41 patients, 38 of whom achieved complete remission. In the follow-up period additional Hpr measurements were taken from 17 patients. Three of them eventually relapsed, and showed increased Hpr levels at the time of relapse. Hpr levels remained low in 11 of 14 patients who maintained complete remission, and increased in three. In conclusion, serum Hpr is a new serum tumor marker of potential use in the clinical setting of lymphoma. This work is dedicated to the memory of Dr. Arie H. Bartal, a dedicated oncologist and friend. This work was supported by Chemotech Thechnologies Ltd., by grant no. 3676 from the Chief Scientist’s Office of the Ministry of Health, Israel, and by the Fund for Promotion of Research in the Technion.     

CYFRA21—1,CEA联合测定对癌性胸水的诊断价值

检测１１８例胸水ＣＹＦＲＡ２１－１和ＣＥＡ浓度以及ＡＤＡ活性。其中癌性不６７例，结核性胸水４０例，其它良性渗漏液１１例。ＣＹＦＲＡ２１－１对癌胸水的敏感度为６２．６％，特异度为９０．２％，ＣＥＡ的敏感度为５５．２％，特异度为１００％。联合测定ＣＹＦＲＡ２１－１和ＣＥＡ对癌性胸水的敏感度可提高到８５．１％，其临床应用值值得重视。