Molecular diagnosis and treatment for esophageal carcinoma with p53

We reviewed molecular diagnosis and molecular treatment using p53 as a target for esophageal squamous cell carcinoma (SCC). First, we analyzed serum p53 antibodies (s-p53 Abs) in patients with esophageal SCC. Positive rate was 31% in all patients analyzed (n = 292) and 23% in stage I patients (cTNM/UICC stages) (n = 48). Presence of s-p53 Abs was significantly associated with p53 protein overexpression in resected tumor specimens. Seropositive patients were more likely to be resistant to chemoradiation and had a worse prognosis than seronegative patients. Second, we performed a clinical study of p53 gene therapy in 10 patients with unresectable chemoradiation-resistant esophageal SCC. In 9 patients, stability of the local tumor was achieved. No serious adverse events related to Ad5CMV-p53 have occurred in these patients, and the trial was safely conducted. Thus, intratumoral injection of Ad5CMV-p53 is safe, feasible, and biologically active when administered in multiple doses to patients with esophageal cancer. Observations from these clinical studies indicate that p53 is a useful molecular target in the diagnosis and treatment of esophageal SCC. Proceeding of a symposium at the 60th annual meeting of the Japan Esophageal Society: “Clinical aspects of molecular biology for the diagnosis and treatment of esophageal carcinoma.”     

河南安阳地区p53基因第72密码子多态性与HPV相关食管癌的研究

目的探讨河南安阳地区p53基因第72密码子多态性与人乳头瘤病毒(HPV)相关食管癌的关系.方法收集安阳肿瘤医院食管癌病例110例,用PCR方法检测HPV,PCR-RFLP方法分析p53基因第72密码子多态性,病例对比研究方法分析在食管癌病例中HPV感染与p53基因第72密码子多态性的关系.结果 PCR检测结果表明,河南安阳地区食管癌组织中HPV检出率为59.1%;HPV阳性者中,Arg/Arg基因型的频率是60.0%,HPV阴性者中仅为17.8%,两者有统计学显著性(P＜0.05).结论 p53基因第72密码子多态性可能是安阳地区HPV相关食管癌的易感因素之一,携带p53Arg/Arg基因型的个体更容易发生HPV相关的食管癌.     

结肠癌合并血吸虫感染患者癌细胞p53基因的表达

目的 研究原发性结肠癌细胞p53基因mRNA的表达水平及合并血吸虫感染后的差异,探讨其与患者临床病理特征的关系。 方法 38例原发性结肠癌患者分为两组,A组（合并血吸虫感染）20例和B组（未合并血吸虫感染）18例。应用实时荧光定量 PCR和相对定量分析法检测患者肿瘤组织中的p53 mRNA。 结果 p53 mRNA在两组中均可检出,A组中的基因表达水平显著高于B组（P＜0.05）。p53基因mRNA表达水平与年龄、性别相关无显著性,与肿瘤大小、有无淋巴结转移相关具有显著性。结论 p53基因mRNA的高水平表达与结肠癌的侵袭和发展具有显著性差异,血吸虫感染可能对结肠癌患者p53基因的突变有一定影响。     

p53 Mutations in human cholangiocarcinoma: a review.

The reported mortality from intrahepatic bile duct tumours is increasing markedly in industrialised countries, for reasons that remain unknown. Inactivation of the tumour suppressor gene p53, is the commonest genetic abnormality in human cancer and has been implicated in the genesis of cholangiocarcinoma in various immunohistochemical and molecular epidemiological investigations, including gene sequencing studies. The structure and function of p53 and its role in linking cancer to specific carcinogens by way of mutational signatures is reviewed. The findings of previous p53 studies and their relevance in human cholangiocarcinoma are summarised.     

p53与人类肺癌关系研究的进展

p53基因是目前人类所发现的与肿瘤发病相关性最大的抑癌基因(tumorsuppressorgene,TSG)。p53基因的点突变、缺失及灭活在肺癌的发生和进展过程中起着关键性的作用。随着各项研究的不断进展以及高新检测技术的出现,p53在肺癌的早期诊断中的价值得到进一步的肯定。同时,利用p53进行基因治疗及评价肺癌患者预后的研究也逐渐开展起来并取得一定成效。因此深入探讨p53基因与人类肺癌的关系具有重大的意义。本文就p53与人类肺癌关系研究的进展作一论述。     

微囊藻毒素LR促肝癌过程中对p53基因突变与表达的影响

目前所检测到的微囊藻毒素(microcystin,MC)异构体将近80种[1]。其中,微囊藻毒素LR (MCLR)因其毒性大、分布广,已成为目前研究热点,但对其致毒机制,尤其致肝肿瘤机制尚不清楚。我们根据二阶段致癌理论建立MCLR中期动物模型,并分析与细胞周期密切相关的基因p53、p21waf1基因在MCLR促肝癌过程的变化,为更深入了解MCLR的促癌机制提供帮助。     

重组人p53腺病毒感染对携带不同p53状态胃癌细胞增殖和凋亡的影响

目的研究重组人p53腺病毒感染不同p53状态胃癌细胞对其p53蛋白表达、生长抑制率、细胞周期与凋亡率的影响。方法不同浓度重组人p53腺病毒感染3种不同p53状态胃癌细胞,即含野生型p53基因的细胞(wild-type)、含突变型p53基因的细胞(mutant-type)、含空载质粒即p53基因缺失的细胞(vector-cell)。48 h后,用Western blotting法检测p53蛋白在3种胃癌细胞中的表达;用MTT法测定重组人p53腺病毒感染3种胃癌细胞的生长抑制率,用流式细胞仪检测细胞周期分布和凋亡率。结果rAd-p53感染3种胃癌细胞48 h后p53蛋白表达阳性,对照组p53基因缺失的胃癌细胞无表达,对照组含野生型p53基因的细胞和含突变型p53基因的细胞弱表达。rAd-p53对3种胃癌细胞的生长抑制效应在一定的浓度范围内呈剂量依赖性,而与细胞内在的p53状态无关。含野生型p53基因的细胞、含突变型p53基因的细胞和p53基因缺失的细胞感染rAd-p53后诱导G2/M期阻滞与细胞凋亡率分别增加2.5、3.6、3.2倍。结论腺病毒介导p53基因感染3种不同p53状态胃癌细胞改变细胞内在的p53状态,p53蛋白表达、生长抑制率、细胞周期分布、凋亡率均与细胞内在的p53状态无关。     

食管癌组织芯片p53、p16和环氧合酶-2表达的研究

目的食管癌的发生发展是多步骤、多基因变化的演化过程，本研究利用高通量的组织芯片技术，对食管癌组织及癌旁组织的p53、p16和环氧合酶（COX）-2蛋白异常表达进行分析，探讨其相关性及临床意义。方法利用组织芯片技术结合免疫组化法检测86例食管癌组织、40例癌旁组织中p53、p16、COX-2蛋白的表达。结果食管癌组织中p53、COX-2的阳性表达率均显著高于癌旁组织（P〈0．05）。食管癌组织中p16阳性表达率为5．81％，癌旁组织中没有发现p16蛋白表达，差异无统计学意义。p53与p16、p53与COX-2、p16与COX-2蛋白表达均存在差异（P〈0.05）。p53或COX-2表达阳性时组织芯片病理类型为癌性的概率增加，但p16、p53和COX-2三者不存在交互作用。结论p53、COX-2对预测和早期诊断食管癌具有重要意义。     

Expression of p53, p16 and cyclooxygenase-2 in esophageal cancer with tissue microarray

OBJECTIVE: The aim of this study was to obtain a comprehensive survey on the expression of p53, p16 and cyclooxygenase‐2 (COX‐2) in esophageal cancer progression and their clinical significance. METHODS: A tissue microarray containing 86 specimens from esophageal cancer and 40 specimens from adjacent non‐cancer tissue was constructed to survey the expression of p53, p16 and COX‐2 by immunohistochemistry. The influence of each biomarker on the histotype of esophageal lesion was assessed by logistic regression analysis. RESULTS: The expression of p53 and COX‐2 was significantly higher in tumorous tissue than in non‐tumorous tissue. As to p16, no significant difference was detected between tumorous and non‐tumorous tissue. A significant correlation was observed among p53, COX‐2 and p16 expression. Logistic regression analysis revealed that the risk factors of a tumorous histotype were the positive expression of p53 (odds ratio [OR] = 18.214) or COX‐2 (OR = 42.703), and no reciprocal relationship to neoplastic progression was recognized with p53, p16 and COX‐2. CONCLUSIONS: p53 and COX‐2 were independent predictors in esophageal carcinogenesis. Esophageal tissue with a positive expression of p53 or COX‐2 was more likely to develop esophageal cancer.     

p 53 gene rearrangements in chronic myelocytic leukemia

Summary We performed Southern-blot analysis of the p 53 gene in 41 consecutive patients with typical chronic myelocytic leukemia (CML). In two of them, we were able to study cells during both the chronic and the accelerated phases. Only one of the 29 chronic-phase samples had rearrangement of the p 53 gene, whereas three of the nine accelerated-phase samples and one of the five patients in blast crisis exhibited rearrangements. Gene deletion was observed in two patients, one in accelerated phase and the other in blast crisis. One patient with a nonrearranged p 53 gene in chronic phase showed rearrangement after progression to the accelerated phase. On the other hand, one patient in accelerated phase exhibited rearrangements which disappeared after reversion to chronic phase with successful treatment. Our findings support the opinion that alterations of the p 53 gene may play an important role in CML evolution.     

食管癌组织中VEGF和P53及血管密度的研究

目的　探讨VEGF、p5 3及血管密度 (MVD)在食管鳞癌组织中的表达及其与预后的关系。方法　应用免疫组化技术检测食管鳞癌组织中VEGF、p5 3及MVD的表达。结果　 41例VEGF阳性病例MVD平均为 5 2 3 5± 14 13 ,2 4例VEGF阴性病例MVD平均为 40 67± 10 2 7,VEGF表达与MVD呈正相关 ,P <0 0 5。在 3 9例p5 3蛋白阳性病例中VEGF阳性表达 3 0例 ,在 2 6例p5 3蛋白阴性病例中VEGF阳性表达 12例 ,p5 3蛋白阳性表达与VEGF表达呈正相关 ,P <0 0 5。 3 9例p5 3阳性病例MVD平均为 5 3 3 8± 12 16,2 6例p5 3蛋白阴性病例MVD平均为 41.2 7± 11.67,两者差异具有显著意义 ,P <0 0 5。 3 8例生存期小于 5年的患者VEGF阳性 2 7例、p5 3阳性 2 6例、MVD平均为 5 0 12± 10 64 ,2 7例生存期 5年以上病例VEGF阳性 14例、p5 3阳性 13例、MVD平均为 3 9 72±12 67,VEGF、p5 3、MVD与生存期长短呈负相关 ,P <0 0 5。VEGF、p5 3、MVD在食管鳞癌中的表达存在有异质性。结论　VEGF、p5 3、MVD与食管癌血管生成密切相关 ,可作为食管癌预后判断的一个重要指标     

端粒酶催化亚单位和p53基因在大肠癌中表达的研究

目的　研究端粒酶催化亚单位 (hTERT)和p5 3基因在大肠癌组织中的表达 ,进而探讨大肠癌发生过程中端粒酶活化的分子机制。方法　分别采用原位逆转录聚合酶链反应 (RT PCR)和端粒重复序列扩增 (TRAP)法检测 46例大肠癌及其相应正常组织中hTERTmRNA表达与端粒酶活性。用免疫组化S P法测定上述组织标本中的p5 3蛋白表达。结果　hTERTmRNA与端粒酶活性在大肠癌组织中阳性表达率分别为 87 0 % (4 0 /4 6)和 80 4% (3 7/4 6) ,均明显高于相应正常组织 (P <0 .0 1)。大肠癌中hTERYmRNA表达与端粒酶活性明显相关 (r=0 .696,P <0 .0 1)。p5 3蛋白在大肠癌中阳性表达率为 67 4% (3 1/4 6) ,而正常组织未见阳性表达 ,二者比较差异有显著性 (P <0 .0 1)。大肠癌组织p5 3蛋白表达与hTERTmRNA表达或端粒酶活性无显著相关性 (P >0 .0 5 )。结论　端粒酶激活与大肠癌的发生密切相关 ,hTERTmRNA表达上调可能在大肠癌端粒酶活性调节中发挥重要作用。p5 3蛋白表达增强可能与端粒酶激活无直接相关     

肝细胞癌基因P53异常的研究

目的：分析我国重庆地区肝细胞癌Ｐ５３基因失活机制及突变谱。方法：采用ＰＣＲ—ＲＦＬＰＰＣＲＳＳＣＰ和ＰＣＲ直接测序技术对来自我国重庆地区２８例肝细胞癌Ｐ５３抑癌基因结构异常进行了分析。结果：６１．５１％的肝癌存在ｐ５３的杂合缺失：５０％肝癌伴有ｐ５３基因突变，其突变模式为突普通散在于５６７和８外显子，其中第７外显子２４９们密友情子突弯率最高（２１％）；具有突变的肝癌多同时伴夺缺失。伴有ｐ５３基因结构异常的肝癌均属进展期。结论：我国重庆地区肝癌存在Ｐ５３基因结构异常，Ｐ５３基因结构异常，ｐ５３基因突变模式反映了该地区肝癌发生可能与肝炎病互和黄贡互素两种因素及其相互作用有关；ｐ５３基因结构异常属肝癌晚期事件，可能参加与肝癌的进展过程。     

Adenovirus mediated p53 tumour suppressor gene therapy for human gastric cancer cells in vitro and in vivo

BACKGROUND/AIMS: Gastric cancer is one of the most prevalent forms of cancer in East Asia. Point mutation of the p53 gene has been reported in more than 60% of cases of gastric cancer and can lead to genetic instability and uncontrolled cell proliferation. The purpose of this investigation was to evaluate the potential of p53 gene therapy for gastric cancer. METHODS: The responses of human gastric cancer cell lines, MKN1, MKN7, MKN28, MKN45, and TMK-1, to recombinant adenoviruses encoding wild type p53 (AdCAp53) were analysed in vitro. The efficacy of the AdCAp53 treatment for MKN1 and MKN45 subcutaneous tumours in nude mice was assessed in vivo. RESULTS: p53-specific growth inhibition was observed in vitro in two of four gastric cancer cell lines with mutated p53, but not in the wild type p53 cell line. The mechanism of the killing of gastric cancer cells by AdCAp53 was found, by flow cytometric analysis and detection of DNA fragmentation, to be apoptosis. In vivo studies showed that the growth of subcutaneous tumours of p53 mutant MKN1 cells was significantly inhibited by direct injection of AdCAp53, but no significant growth inhibition was detected in the growth of p53 wild type MKN45 tumours. CONCLUSIONS: Adenovirus mediated reintroduction of wild type p53 is a potential clinical utility in gene therapy for gastric cancers.     

河南食管癌高低发区人群食管上皮p53变化和细胞增生状况的比较研究

目的探讨食管癌高、低发区人群食管上皮肿瘤抑制基因ｐ５３的变化及其与细胞增生状况的关系，以了解食管癌变的分子学基础．方法食管癌高发区居民２２０例和低发区居民５０例分别进行食管内镜检查、粘膜活检和组织病理学检查．根据细胞和组织形态结构区分为正常、基底细胞增生和间变，并采用免疫组化ＡＢＣ法分析各级病变和正常上皮Ｐ５３蛋白及细胞增生核抗原（ＰＣＮＡ）的改变，并计数上皮单位面积内Ｐ５３和ＰＣＮＡ阳性细胞数，进行定量比较研究．结果河南高、低发区人群食管上皮细胞均出现不同程度的Ｐ５３和ＰＣＮＡ的变化．高、低发区居民食管正常上皮和病变组织ＰＣＮＡ免疫阳性细胞数和高发区Ｐ５３阳性细胞数随病变加重呈升高趋势．但是，低发区居民食管上皮Ｐ５３阳性细胞数却无此现象．结论肿瘤抑制基因ｐ５３的变化和ＰＣＮＡ改变有明显关系，均随病变的进展而升高．高发区居民食管各级病变Ｐ５３改变明显高于低发区居民，此结果有助于对食管癌地域性分布差异的分子学基础的了解．     

Differential expression of retinoic acid receptors and p53 protein in normal, premalignant, and malignant esophageal tissues

Purpose: Esophageal cancer remains a significant health problem worldwide. The very low 5-year survival rates and rapid increase in the incidence of adenocarcinoma indicate the urgent need for early identification of and new approaches to the prevention and treatment of this cancer. Methods: To find biomarkers for early identification of the disease, we analyzed nuclear retinoic retinoid receptor mRNAs, p53 protein, and the proliferation marker Ki 67 in surgical specimens of normal, mildly, and severely dysplastic and malignant esophageal tissues. Results: Nuclear retinoid receptors (RAR-α, RAR-γ, and RXR-α) were expressed in most (79%–100%) normal, dysplastic, and malignant esophageal mucosae, whereas expression of RAR-β was progressively lost from normal esophagus to carcinoma (84%–54%). In contrast, expression of p53 protein and Ki 67 were dramatically increased in severely dysplastic and cancerous tissues of the esophagus (from 5% to 62%). Conclusions: Loss of RAR-β expression and accumulation of p53 and Ki 67 proteins may serve as biomarkers for esophageal cancer. Received: 17 April 2000 / Accepted: 12 July 2000     

Alterations in p53 expression and cell proliferation in esophageal epithelia among patients from geographical areas with high or low incidence of esophageal cancer

AIM: To study changes in p53 expression and cell proliferation in esophageal epithelia of subjects from high or low esophageal cancer incidence areas in Henan Province to understand their molecular basis.METHODS: Esophageal endoscopic mucosa biopsies were acquired and histopathological examinations were performed on 220 subjects from high esophageal cancer incidence areas and 50 subjects from low incidence areas in Henan Province. Esophageal epithelia were diagnosed as normal, basal cell hyperplasia or dysplasia based on cell morphology and tissue structure. Immunohistochemistry avidin biotin peroxidase complex (ABC method) was performed to analyze alterations in p53 and proliferating cell nuclear antigen (PCNA) expression in normal epithelia and epithelia with different lesion severities. The numbers of p53-positive and PCNA-positive cells were counted.RESULTS: p53- and PCNA-positive nuclei were present in esophageal epithelia from subjects from both high and low incidence areas. The number of PCNA-positive cells gradually increased with lesion severity for both the high and low incidence areas. The number of p53-positive cells was higher in high incidence areas compared to low incidence areas, and rapidly increased with lesion severity. p53 expression positively correlated with PCNA expression.CONCLUSION: The number of both p53- and PCNA-positive cells increased with lesion severity. p53 expression was higher in subjects from high esophageal cancer incidence areas compared to those from low incidence areas. These results may shed light into the molecular basis for the geographical distribution of esophageal cancer.     

抑癌基因p53及p16单独或共转染治疗非小细胞肺癌的动 …

目的 观察抑癌基因ｐ５３和ｐ１６单独或或共转染在动物整体中治疗非小细胞肺癌（ＮＳＣＬＣ）的效果。方法 采用培养细胞移植法,将２ ＮＳＣＬＣ细胞系Ａ５６９接种于裸小鼠背部皮下,建立裸小鼠皮下肺癌移植瘤模型。将２５只荷瘤裸小鼠随机分成空白对照组、十八酰基胺阳离子（ＳＡ０对照组、ｐ５３基因组、ｐ１６基因组、ｐ５３＋ｐ１６基因组,共５组,每组５只。采用瘤体内直接注射的方法,用ＳＡ脂质体介异,将ｐ５３和（或     

肺癌患者肺组织和痰标本p53基因突变检测及其临床意义

目的　探讨肺癌患者支气管肺活检组织和痰标本中p5 3基因突变检测在肺癌诊断中意义。方法　应用聚合酶链反应 (PCR) 单链多肽性 (SSCP) 银染法检测 12 0例肺癌和 40例良性肺疾病支气管肺活检组织和痰标本p5 3基因第 5～ 8外显子突变情况。结果　 60例肺癌患者癌组织、癌旁组织和对侧支气管组织p5 3基因突变检出率分别为 60 % (3 6/ 60 )、17% (10 / 60 )和 5 % (3 / 60 ) ,三组间比较差异有显著性 (P <0 .0 0 5 )。 2 0例良性肺疾病支气管肺组织p5 3基因突变检出率为 5 %。故活检组织p5 3基因突变检出肺癌的敏感性为 60 % ,特异性为 95 % ;另外 60例肺癌痰标本p5 3基因突变检出的敏感性为 43 % (2 6/ 60 ) ,特异性为 10 0 %。肺癌痰标本p5 3基因突变联合细胞学检测使肺癌的诊断率从单纯细胞学的 48% (2 9/ 60 )提高到 68% (41/ 60 )。结论　痰标本p5 3基因检测联合细胞学检查可提高肺癌的诊断率     

Down-expression of tumor protein p53-induced nuclear protein 1 in human gastric cancer

AIM: Overexpression of tumor protein p53-induced nuclear protein 1 (TP53INP1) induces G1 cell cycle arrest and increases p53-mediated apoptosis. To clarify the clinical importance of TP53INP1, we analyzed TP53INP1 and p53 expression in gastric cancer. METHODS: TP53INP1 and p53 expression were examined using immunohistochemistry in 142 cases of gastric cancer. The apoptosis of gastric cancer cells was analyzed using the TUNEL method. The relationship between the expression of TP53INP1 and clinicopathological factors was statistically analyzed. RESULTS: TP53INP1 was expressed in 98% (139/142 cases) of non-cancerous gastric tissues and was down-expressed in 64% (91/142 cases) of gastric cancer lesions from the same patients. TP53INP1 expression was significantly decreased (43.9%) in poorly differentiated adenocarcinoma compared with well or moderately differentiated adenocarcinoma (81.6%). Cancers invading the submucosa or deeper showed lower positively (59.1%) compared with mucosal cancers (85.2%). Decrease or loss of TP53INP1 expression was significantly correlated with lymphatic invasion (54.3% vs 82.0% without lymphatic invasion) and node-positive patients (31.3% vs 68.3% in node-negative patients). P53 was expressed in 68 (47.9%) patients of gastric cancer, whereas it was absent in normal gastric tissues. A significant association was also observed between TP53INP1 status and the level of apoptosis in tumor cells: the apoptotic index in TP53INP1-positive tissues was significantly higher than that in TP53INP1-negative portions. Finally, when survival data were analyzed, loss of TP53INP1 expression had a significant effect in predicting a poor prognosis (P=0.0006). CONCLUSION: TP53INP1-positive rate decreases with the progression of gastric cancer. TP53INP1 protein negativity is significantly associated with aggressive pathological phenotypes of gastric cancer. TP53INP1 is related to the apoptosis of gastric cancer cells. The decreased expression of the TP53INP1 protein may reflect the malignant grade of gastric cancer and is regarded as an adverse prognostic factor.