利用组织芯片技术分析肝细胞癌组织P21WAF1/CIP1、PCNA和Ki-67的表达及意义

利用组织芯片技术检测P21WAF1/CIP1、PCNA与Ki-67在肝细胞癌组织中的表达与分布,初步探讨p21 1WAF1/CIP1、PCNA与Ki-67在肝细胞癌发生机制以及预后评估方面的作用.采用免疫组化技术结合临床病理资料检测了61例肝癌和50例癌旁组织的P21WAF1/CIP1、PCNA、Ki-67的表达情况.PCNA和Ki-67表达与肝癌的分化程度相关,PCNA与肝癌的侵袭转移潜力有关(P＜0.05),p21WAF1/CIP1在实验中未见有明显差异,但在表达程度与肝癌组织的分化程度之间亦存在着一定的相关关系.应用组织芯片高效检测临床组织样本具有快速、方便、经济、准确的优点.P21WAF1/CIP1、PCNA与Ki-67的异常表达可能与肝癌的发生发展密切相关.     

Role of p53 and p21/WAF1 detection in patient selection for preoperative radiotherapy in rectal cancer patients

BACKGROUND: Recent studies showed that p53 and p21 may play major roles in determining tumor radiosensitivity through the apoptosis pathway. The aim of this study was to investigate the predicting value of radiosensitivity in human rectal carcinoma. METHODS: p53 and p21/WAF1 expressions in formalin fixed, paraffin-embedded, preradiation biopsy samples from 49 patients with primary rectal carcinoma were analyzed immunohistochemically. p53 and p21 expressions and their relationships with histopathologic changes after radiation and other clinical features were evaluated. RESULTS: Expressions of p53 and p21/WAF1 were 49 and 28.6 percent, respectively. In 36.7 percent of total tumors, significant histopathologic effect can be observed. There was a significant inverse expression of p53 and p21. Most of the p53(+) or p21(–) tumors were radioresistant, and the majority of p53(–) or p21(+) tumors were radiosensitive. Tumors size in the radiosensitive, p53(–), or p21(+) group decreased more significantly than in radioresistant, p53(+), or p21(–) group (P<0.01), and patients with radioresistant, p53(+), or p21(–) tumors had more local recurrence, more distant metastasis, and a shorter five-year survival rate than those with radiosensitive, p53(–), or p21(+) tumors, but without statistic significance. No statistically significant correlation can be observed between other tumor clinical features and radiosensitivity, p53, or p21 expressions. CONCLUSION: Immunohistochemistry detection of p53 and p21 expressions may be useful parameters for more radiosensitive patients selected for preoperative radiotherapy.Supported by a Grant-in-aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.Presented at the meeting of the Asian-Pacific Congress of Gastroenterology, Yokohama, Japan, September 19 to 23, 1996.     

肝胆管癌丙型肝炎病毒感染与p53和p21WAFD/CIP1异常表达的关系

为探讨丙型肝炎病毒（ＨＣＶ）感染与Ｐ５３和Ｐ２１ＴＷＡＦ－／ＣＩＰ１基因的关系。采用 化技术对２９例原发性肝胆管癌中ＨＣＶ抗原（ＮＳ５－Ａｇ）、ｐ５３和ｐ２１＾ＷＡＦＩ＝／ＣＩＰ１蛋白表达进行研究。结果：２９例胆管癌中ＮＳ５－Ａｇ、ｐ５３及Ｐ２１ＴＷＡＦＩ／ＣＩＰＩ蛋白表达进行研究。结果：２９例胆管癌中ＮＳ５－Ａｇ、Ｐ５３display structure     

Expression and prognostic relevance of p21WAF1 in stage III esophageal squamous cell carcinoma

The prognostic effect of p21^WAF1 expression on esophageal squamous cell carcinoma patients is controversial. Further clarifying the effect of this protein is beneficial for optimizing the patient outcomes. In the current study, we investigated the expression of p21^WAF1 protein in 189 specimens of stage III ESCC by immunohistochemistry. As shown by the Kaplan–Meier curve, the overall survival rate of the positive‐expression group was significantly higher than that of the negative‐expression group (P < 0.05). No significant correlation was observed between p21^WAF1 expression and clinicopathological parameters in terms of gender, age, tumor location, tumor grade, pathological stage, and number of regional lymph node metastases (P > 0.05). We concluded that p21^WAF1 played an intricate role in the tumorigenesis and development of ESCC. p21^WAF1 could serve as a positive prognostic predictor for stage III ESCC patients.     

p21~(WAF1/CIP1/SDI1)在大鼠肾脏组织的增龄性变化及意义

目的 研究p21野生型p53活化片段1/细胞周期蛋白依赖性激酶影响蛋白1/衰老细胞衍生抑制剂1(p21WAF1/CIP1/SDI1)在大鼠肾脏中随年龄增长的表达变化规律. 方法 取3月龄、12月龄及24月龄健康雄性Wistar大鼠肾组织进行衰老相关β-半乳糖苷酶(SA-β-gal)活性染色,TUNEL法检测细胞凋亡,采用逆转录多聚酶链反应(RT-PCR)和Western印迹法(Western blot assay)分别在基因及蛋白质表达水平上检测肾脏组织中p21WAF1/CIP1/SDI1的表达变化,并用免疫组化法检测p21WAF1/CIP1/SDI1在肾脏组织中的表达与定位. 结果 大鼠肾脏组织SA-β-gal活性随年龄增长逐渐增强,凋亡细胞也随年龄增长逐渐增加(P＜0.05);p21WAF1/CIP1/SDI1 mRNA表达随年龄增长逐渐增强,不同月龄比较差异有统计学意义(P＜0.05).Western印迹亦显示p21WAF1/CIP1/SDI1蛋白表达随鼠龄增加逐渐增强(P＜0.05).免疫组化结果显示,p21WAF1/CIP1/SDI1蛋白表达于大鼠肾小球足细胞,其在肾小管与间质细胞中也有表达,且随年龄增长表达增加(P＜0.05). 结论 p21WAF1/CIP1/SDI1在大鼠肾脏组织中的表达随年龄增加而增强,可作为肾脏组织中重要的衰老指标.     

P21/WAF1蛋白在肝细胞癌中的表达及DNA 含量分析的意义

目的　探讨肝细胞癌p2 1/WAF1蛋白的表达情况及DNA含量的定量分析及临床意义。方法　应用免疫组织化学和多功能真彩色图像分析仪检测 60例肝细胞癌 p2 1蛋白表达量和DNA含量 ,同时取 2 0例癌旁组织作对照。结果　 60例肝细胞癌 p2 1蛋白阳性表达率为 47% (2 8/60 ) ,癌旁组织为 15 % (3 /2 0 )。 60例肝细胞癌中高分化癌阳性表达率为 13 % (2 /15 ) ,低分化癌为 5 8% (2 6/4 5 )。可见p2 1蛋白在肝细胞癌中的表达明显高于癌旁肝组织 (P <0 .0 5 ) ,低分化癌明显高于高分化癌 (P <0 .0 1)。肝细胞癌DNA相对倍体均值 (U值 )范围为 1.0 8～ 3 .17,U值随病理分级的增高而增高 (P <0 .0 5 )。结论　p2 1蛋白的表达情况与肝细胞癌的发生发展及恶性程度有关 ,肝细胞癌DNA含量与病理学特征及临床生物学行为之间有密切的关系。两者相结合可作为肝细胞癌诊断和预后的参考指标。     

KLF6、p21^WAF1/CIP1、CyclinD1在结直肠癌中的表达及意义

目的研究转录因子KLF6、p21WAF1/C IP1及Cyc linD1在结直肠癌中的表达及意义。方法应用免疫组化Envision法对58例结直肠癌组织、20例结直肠黏膜慢性炎症组织中的KLF6、p21WAF1/C IP1及Cyc linD1蛋白表达进行检测。结果结直肠癌组织KLF6、p21WAF1/C IP1及Cyc linD1蛋白表达率均与结直肠黏膜慢性炎症组织比较有统计学差异（P〈0.05）;KLF6、p21WAF1/C IP1及Cyc linD1蛋白在结直肠癌组织中的表达与其浸润深度及预后有关（P〈0.05）。结论 KLF6、p21WAF1/C IP1及Cyc linD1在结直肠癌的发生发展中可能起重要作用。     

HnRNP A2/B1及p21WAF1蛋白在非小细胞肺癌中表达及意义

目的探讨核内不均一核糖核蛋白A2/B1（hnRNP A2/B1）和p21WAF1蛋白在人非小细胞肺癌（NSCLC）组织中的表达,并进行相关性分析。方法采用免疫组化S-P法检测53例NSCLC组织、40例癌旁组织、7例肺良性病变组织中hnRNPA2/B1和p21WAF1表达。结果 NSCLC组织中hnRNP A2/B1和p21WAF12种蛋白的阳性表达率分别为66.0%和45.2%,显著高于癌旁肺组织和肺良性病变肺组织,差异有统计学意义（P〈0.05）。它们均与分化程度、淋巴结转移有关（P〈0.05）,与组织类型、临床分期无关（P〉0.05）。两种蛋白在NSCLC表达呈显著负相关（P=0.001）。结论 hnRNP A2/B1可能通过下调p21WAF1的表达而缩短细胞周期,促进NSCLC的癌细胞增殖。     

p21WAF1/CIP1 基因转染对胃癌细胞生物学活性的影响

目的探讨p21^WAF1／CIP1基因(p21基因)对人胃癌细胞系(BGC)生物学活性的影响。方法应用分子克隆技术构建p21^WAF1／CIP1基因真核表达载体，然后用脂质体法将其导入到人胃癌细胞BGC中，经G418筛选获得可稳定表达p21^WAF1／CIP1的人胃癌细胞克隆，用中性红摄入法观察细胞生长速率，流式细胞仪(FcM)检测细胞周期变化。结果p21导入胃癌BGC细胞后，肿瘤细胞增值能力明显受到抑制，并出现细胞周期G1期阻滞。结论p21基因具有抑制胃癌细胞增殖的作用，可作为胃癌基因治疗的靶基因．     

p21<Superscript>WAF1/CIP1</Superscript> is more effective than p53 in growth suppression of mouse renal carcinoma cell line Renca in vitro and in vivo

Purpose Although there are many controversial reports about the effect of p53 and p21^WAF1/CIP1 overexpression in different human tumor cells, the p53 gene is shown to be a more effective candidate for cancer gene therapy because of its more pronounced ability to induce apoptosis. In the present study, we present the effect of p53 and p21^WAF1/CIP1 overexpression on mouse renal carcinoma cells in vitro and in vivo.Methods p53 and p21^WAF1/CIP1 genes were introduced into Renca cells using adenoviral vectors (Ad5CMV-p53 and Ad5CMV-p21). The induction of apoptosis was measured using Annexin V assay and DNA fragmentation analysis. The expression of proteins was examined using immunocytochemistry and Western blot methods. The ability of adenoviral vectors to inhibit tumorigenicity of Renca cells, as well as the growth of pre-established tumors was measured.Results In vitro growth assays revealed higher growth suppression after Ad5CMV-p21 infection. Although both vectors induced apoptosis, Ad5CMV-p53 was slightly more efficient. In vivo studies in Balb/c mice, demonstrated that tumorigenicity was completely suppressed by Ad5CMV-p21. Besides this, Ad5CMV-p21 significantly inhibited the growth of established tumors, while Ad5CMV-p53 did not.Conclusions These data suggest that p21^WAF1/CIP1 is a more potent growth suppressor than p53 of mouse tumor cells Renca. The divergent responses of tumor cells to p21^WAF1/CIP1 overexpression could be due to various networks that differ between species.     

术前中性粒细胞与淋巴细胞比值对行根治性切除术的肝细胞癌患者预后的影响

目的评价术前中性粒细胞与淋巴细胞比值(NLR)对经外科根治性切除的肝细胞癌(HCC)患者预后的影响及其预测价值。方法回顾性分析245例2004年1月1日-2009年12月31日在中国人民解放军总医院接受根治性切除术的HCC患者的临床资料。通过Log-rank单因素分析法评估各临床病理特征包括NLR对总生存时间的影响;采用Cox比例风险模型将有意义的变量进一步行多因素分析。结果以NLR值为1.5、2及3作为分界点,将研究对象分为4组,分别为NLR1.5、1.5≤NLR2、2≤NLR3及NLR≥3,各组中位总生存时间分别为39.6、38.3、25.4和19.9个月,差异具有统计学意义(P=0.003)。多因素分析显示术前外周血NLR、AFP、ALT水平、肿瘤结节数量、肿瘤组织最大径以及门静脉癌栓是影响HCC患者预后的独立危险因素(P值均0.05)。结论术前外周血NLR的水平可以作为外科切除术后HCC患者的新的预后标志物。     

Immunohistochemical analysis of tumor biological factors in hepatocellular carcinoma: relationship to clinicopathological factors and prognosis after hepatic resection

Background The relationship between patient prognosis and various tumor biological factors has been reported previously, and prognostic factors of tumor biology may improve predictions of prognosis after hepatectomy for hepatocellular carcinoma (HCC) and may contribute to a new staging classification. This study was designed to provide an immunohistochemical analysis of tumor biological factors in patients who underwent hepatectomy for HCC.Methods Factors analyzed included p53 overexpression, microvessel counts, proliferating cell nuclear antigen, and expression of nm23. We examined 81 HCCs from patients with chronic liver diseases.Results In patients who underwent chemoembolization before surgery, or those a who had confluent multinodular tumor, p53 expression tended to be higher than in patients without chemoembolization (33% vs 11%) or those with a simple nodular tumor (28% vs 10%), but the difference was not statistically significant (P = 0.051 and P = 0.092, respectively). A lower tumor microvessel count and negative nm23 expression were significantly associated with poor disease-free survival by univariate analysis (P 0.01 and P 0.05, respectively). A lower tumor microvessel count was found to be a significant prognostic factor for disease-free and overall survivals (risk ratios, 2.44 and 3.13, respectively; P 0.05), in addition to tumor size, vascular invasion, and longterm ascites, by Coxs multivariate analysis.Conclusions Tumor microvessel count appears to be a useful prognostic marker for predicting HCC recurrence and patient survival.