上皮性卵巢癌组织中TUBB3和ERCC1的表达

目的通过检测TUBB3和ERCC1在上皮性卵巢癌组织中的表达情况,探讨两者与上皮性卵巢癌化疗疗效及预后的关系。方法采用免疫组化技术检测25例正常卵巢组织和50例上皮性卵巢癌组织中TUBB3和ERCC1蛋白的表达情况。结果 TUBB3和ERCC1在上皮性卵巢癌组织中的阳性率明显高于正常卵巢组织（P〈0.05）,两者的阳性表达率与上皮性卵巢癌的分化程度及临床分期有关。上皮性卵巢癌组织中TUBB3和ERCC1蛋白表达呈正相关关系（r=0.332,P〈0.05）。TUBB3、ERCC1阳性表达患者化疗有效率分别为34.6%和31.4%,均明显低于阴性患者的91.7%和93.3%（P〈0.05）。结论上皮性卵巢癌组织中TUBB3、ERCC1的表达可能影响化疗疗效及其预后存,两者阳性表达可能提示上皮性卵巢癌化疗耐药,可作为上皮性卵巢癌化疗耐药和预后检测的重要指标。     

切除修复交叉互补基因1、乳腺癌易感基因1及微管蛋白β3检测在晚期肺鳞状细胞癌患者GP方案治疗中的指导意义

目的 探讨切除修复交叉互补基因1(ERCC1)、乳腺癌易感基因1(BRCA1)及微管蛋白β3(TUBB3)检测在晚期肺鳞状细胞癌患者吉西他滨+顺铂（GP）方案治疗中的指导意义。方法 选取74例行GP方案治疗的晚期肺鳞状细胞癌患者,采用免疫组化法检测ERCC1、BRCA1及TUBB3表达情况。比较不同临床特征晚期肺鳞状细胞癌患者的ERCC1、BRCA1及TUBB3表达情况。根据免疫组化结果对患者进行分组,3阳性为A组（n=16）,2阳性+1阴性为B组（n=18）,1阳性+2阴性为C组（n=19）,3阴性为D组（n=21）,比较4组患者的临床特征、总有效率及疾病控制率。结果 74例晚期肺鳞状细胞癌患者肺鳞状细胞癌组织中ERCC1、BRCA1、TUBB3阳性表达率分别为45.95%、55.41%、47.30%。不同性别、临床分期、分化程度肺鳞状细胞癌患者肺鳞状细胞癌组织中ERCC1、BRCA1及TUBB3阳性表达率比较,差异均有统计学意义（P﹤0.05）。4组患者的性别、年龄、吸烟时间比较,差异均无统计学意义（P﹥0.05）;4组患者的临床分期、分化程度比较,差异均有统计学意义（P﹤0.05...     

ERCC1，TUBB3在上皮性卵巢癌中表达及其临床病理特征和相关性的研究

目的研究核苷酸切除修复交叉互补组1（ERCC1）和TUBB3编码的β-tubulin-Ⅲ（3型B微管蛋白）在卵巢上皮性癌中的表达情况、病理特征及其临床意义,为个体化治疗提供一定的理论依据。方法本研究主要通过逆转录一实时荧光定量反应（RTq-PCR）方法检测上皮性卵巢癌组织中ER．CCI和TUBB3mRNA的表达情况,进一步分析上皮性卵巢癌临床病理特征与ERCC1,TUBB3的关系及二者之间的相关性。结果ERCC1和TUBB3在上皮性卵巢癌中均呈高表达,表达率分别为28．21％（22／78）和29．49％（23／78）。单因素分析ERCC1、TUBB3基因在上皮性卵巢癌高表达患者中仅与残余肿物大小有关（P〈0．05）,二者与其他临床病理因素无关。ERCC1和TUBB3在上皮性卵巢癌中表达水平呈正相关（r=0．4249,P〈0．05）。结论上皮性卵巢癌组织存在不同水平的ERCC1和TUBB3基因mRNA,二者在癌组织中表达密切相关。ERCC1、TUBB3基因在上皮性卵巢癌中的表达与残余肿物大小呈正相关。     

非小细胞肺癌组织中ERCC1及TUBB3基因mRNA的表达及其临床意义

目的:探讨ERCC1及TUBB3基因mRNA在非小细胞肺癌(NSCLC)组织、癌旁组织和良性组织中的表达,两者的关系及其与肺癌患者临床、病理等特征的关系。方法:采用逆转录-聚合酶链反应(RT-PCR)方法检测82例非小细胞肺癌组织、36例癌旁组织及15例良性组织中ERCC1及TUBB3基因mRNA的表达。结果:ERCC1和TUBB3基因在NSCLC患者癌、癌旁及良性组织中均有表达,ERCC1在这三种组织中阳性表达率分别为54.9%(45/82)、44.4%(16/36)及73.3%(11/15),差异无统计学意义(P=0.165);而TUBB3阳性表达率分别为18.3%(15/82)、2.8%(1/36)及93.3%(14/15),三者差异有统计学意义(P=0.000),癌组织表达率18.3%高于癌旁组织2.8%(P=0.048)。ERCC1基因在腺癌表达率63.0%(34/54)高于鳞癌39.3%(11/28)(P=0.041)。TUBB3基因在腺癌表达率25.9%(14/54)高于鳞癌3.6%(1/28)(P=0.013)。但是二者在其余临床和病理等特征方面的表达均无统计学意义。癌组织中ERCC1及TUBB3表达呈中度正相关(r=0.429,P=0.000)。结论:ERCC1及TUBB3基因mRNA在NSCLC中腺癌表达均高于鳞癌,并且二者在癌组织中表达有密切的关系。     

免疫组化和qPT-PCR检测肺癌预后标记物的比较

目的:比较qRT-PCR和免疫组化检测肺癌患者的ERCC1、BRCA1、RRM1、TUBB3的表达状况及与患者预后的相关性。方法:进展期非小细胞肺癌患者取病变穿刺组织同时行qRT-PCR及免疫组化检测,根据检测结果分为高表达组和低表达组,分别观察各组生存时间,以总生存为研究终点。结果:免疫组化检测ERCC1阴性患者的OS优于ERCC1阳性患者,BRCA1阴性患者的OS优于BRCA1阳性患者,RRM1表达情况与预后无差异。TUBB3阴性患者的OS优于TUBB3阳性患者。然而qRT-PCR检测时,ERCC1、BRCA、TUBB3的表达状态与OS、DFS或RR无相关性。结论:进展期非小细胞肺癌ERCC1、BRCA1、RRM1、TUBB3的表达与预后相关,免疫组化检测优于qRT-PCR。     

基因TOPO Ⅱα、TUBB3、ERCC1在小儿肾母细胞瘤组织中的表达及其临床意义

目的 探究基因TOPOⅡα、TUBB3、ERCC1在小儿肾母细胞瘤组织中的表达及其临床意义。方法 选择40例肾母细胞瘤患儿的病理组织标本，另同期采集距离肿瘤组织2～3 cm的癌旁组织标本作为对照组。分析两者拓扑异构酶Ⅱα(TOPOⅡα)、3型β微管蛋白编码基因(TUBB3)、核苷酸切除修复交叉互补基因(ERCC1)表达情况；另分析TOPOⅡα、TUBB3、ERCC1与肾母细胞瘤患儿病理特征的相关性。结果 癌组织中TOPOⅡα、TUBB3、ERCC1高表达率均高于癌旁组织，差异有统计学意义(P<0.05)。TOPOⅡα高表达者病理分期Ⅲ～Ⅳ占比率高于中低表达者，差异有统计学意义(P<0.05);ERCC1高表达者病理分期Ⅲ～Ⅳ占比率高于中低表达者，差异有统计学意义(P<0.05)。结论TOPOⅡα、TUBB3、ERCC1在小儿肾母细胞瘤中的表达水平均高于癌旁组织，其中TOPOⅡα、ERCC1均与疾病病理分期有关，二者在肾母细胞瘤的发生及发展中具有重要作用。     

ERCC1、TYMS、RRM1、TUBB3表达与非小细胞肺癌化疗疗效的相关性和意义

[目的]探讨非小细胞肺癌(NSCLC)术后标本中ERCC1、TYMS、RRM1、TUBB3表达与化疗疗效的相关性.[方法]回顾性分析我院病理确诊的72例NSCLC患者手术切除的肿瘤标本,通过分支DNA-液相芯片法,检测ERCC1、TYMS、RRM1、TUBB3基因mRNA表达水平.[结果]ERCC1、TYMS、RRM1、TUBB3阳性表达率分别为38.9％ (28/72)、62.5％(15/72)、55.6％ (46/72)和47.2％(34/72);4者表达与年龄、性别、吸烟、组织学类型、TNM分期、淋巴结转移均无关(P＞0.05),TYMS、TUBB3在高、中分化组中的阳性表达率显著低于低分化组(P=0.003,P＜0.001).ERCC1、TYMS、RRM1和TUBB3阳性表达的化疗患者1年生存率均显著低于阴性表达者(61.5％vs95.5％,P=0.048;53.5％vs 94.9％,P=0.035;58.6％vs 92.9％,P=0.039;47.8％vs 97.4％,P=0.014).[结论]ERCC1、TYMS、RRM1、TUBB3阴性表达者化疗生存率高.ERCC1、TYMS、RRM1、TUBB3阴性表达可作为NSCLC患者临床化疗受益的指标.     

非小细胞肺癌组织中ERCCl及TUBB3基因mRNA的表达及其临床意义

目的：探讨ERCC1及TUBB3基因mRNA在非小细胞肺癌（NSCLC）组织、癌旁组织和良性组织中的表达,两者的关系及其与肺癌患者临床、病理等特征的关系。方法：采用逆转录-聚合酶链反应（RT-PCR）方法检测82例非小细胞肺癌组织、36例癌旁组织及15例良性组织中ERCC1及TUBB3基因mRNA的表达。结果：ERCC1和TUBB3基因在NSCLC患者癌、癌旁及良性组织中均有表达,ERCC1在这三种组织中阳性表达率分别为54．9％（45／82）、44．4％（16／36）及73．3％（11／15）,差异元统计学意义（P=0．165）;而TUBB3阳性表达率分别为18．3％（15／82）、2．8％（1／36）及93．3％（14／15）,三者差异有统计学意义（P=0．000）,癌组织表达率18．3％高于癌旁组织2．8％（P=0．048）。ERCC1基因在腺癌表达率63．O％（34／54）高于鳞癌39．3％（11／28）（P=0．041）。TUBB3基因在腺癌表达率25．9％（14／54）高于鳞癌3．6％（1／28）（P=0．013）。但是二者在其余临床和病理等特征方面的表达均无统计学意义。癌组织中ERCC1及TUBB3表达呈中度正相关（r=0．429,P=0．000）。结论：ERCC1及TUBB3基因mRNA在NSCLC中腺癌表达均高于鳞癌,并且二者在癌组织中表达有密切的关系。     

ERCC1、RRM1和BRCA1在非小细胞肺癌中的表达及预后意义

目的:探讨DNA修复基因家族成员ERCC1、RRM1和BRCA1在非小细胞肺癌(NSCLC)中的表达及预后意义。方法:应用实时荧光定量PCR技术对32例肺癌及16例癌旁组织中ERCC1、RRM1和BRCA1基因的mRNA进行定量检测。用非参数检验、相关分析、Kap lan-M e ier生存曲线和COX多因素回归分析进行统计分析。结果:NSCLC中ERCC1、RRM1和BRCA1在癌组织内表达量显著高于癌旁组织,且在癌内表达具有正相关性;RRM1在肺鳞癌中高于腺癌,但在不同分期中表达无差异;ERCC1和BRCA1在不同病理类型和分期中表达均无差异;RRM1和BRCA1高表达组的生存期明显长于低表达组;COX多因素回归分析示RRM1表达是影响本组患者预后的独立因素。结论:NSCLC中,ERCC1、RRM1和BRCA1在肺癌组织中的表达显著高于癌旁组织,RRM1和BRCA1高表达组的生存期长于低表达组。RRM1和BRCA1可作为判断预后的一种指标。     

ERCC1、BAG-1 及 BRCA1 基因在非小细胞肺癌中的表达及其与预后的关系

目的 探讨ERCC1、BAG-1和BRCA1基因mRNA在接受以铂类为基础辅助化疗的非小细胞肺癌（non-small cell lung cancer,NSCLC）患者中的表达情况及其与预后的关系。方法 85例NSCLC患者中74例给予以铂类为基础的术后辅助化疗,采集全组患者的肿瘤组织和其中34例患者的癌旁组织,应用半定量RT-PCR法,检测其ERCC1、BAG-1和BRCA1基因mRNA的表达情况,回顾性分析患者的临床病理资料,探讨3个基因的表达与患者总生存期（OS）之间的关系。结果 ①ERCC1、BAG-1阴性表达的患者OS显著长于阳性表达的患者（P均=0.001）。ERCC1、BAG-1阴性表达的患者接受铂类化疗的疗效显著优于阳性表达的患者（P=0.002、P=0.001）。②BRCA1阴性表达的患者OS长于阳性表达的患者,但差异无统计学意义（P=0.057）。③多因素分析显示ERCC1、BAG-1可作为NSCLC患者接受铂类化疗的预后指标（P=0.027、P=0.022）。结论 检测ERCC1和BAG-1的表达可作为指导NSCLC患者术后接受铂类化疗及预后评估的指标。     

mRNA Expression and Clinical Significance of ERCC1, BRCA1, RRM1, TYMS and TUBB3 in Postoperative Patients with Non-Small Cell Lung Cancer

Background: To explore mRNA expression and clinical significance of ERCC1, BRCA1, RRM1, TYMS andTUBB3 genes in tumor tissue of postoperative patients with non-small cell lung cancer (NSCLC). Materialsand Methods: Sixty NSCLC patients undergoing radical operation in our hospital from Nov., 2011 to Jun.,2012 were selected. Plasmid standards of ERCC1, BRCA1, RRM1, TYMS and TUBB3 were established andstandard curves were prepared by SYBR fluorescent real-time quantitative PCR analysis. Samples fromtumor centers were taken to detect mRNA expression of ERCC1, BRCA1, RRM1, TYMS and TUBB3 genesin cancerous tissue during operation. The total mRNA expression quantities were compared according todifferent clinical characteristics. Results: The total expression quantities of 5 genotypes from high to low wereERCC1>RRM1>TUBB3>TYMS>BRCA1 in turn. By pairwise comparisons, other differences showed statisticalsignificance (p<0.05 or p<0.01) except for TYMS and TUBB3 (p>0.05); the low expression rates from high to lowwere ERCC1>TYMS>TUBB3>TUBB3>RRM1>BRCA1 in turn. The expression quantities of BRCA1, RRM1 andTYMS in males, smokers and patients without adenocarcinoma were all significantly higher than that in females,non-smokers and patients with adenocarcinoma, and significant differences were present (p<0.05 or p<0.01).In terms of pathological staging, the expression quantities of BRCA1, RRM1 and TYMS in phases Ⅱa~Ⅱb andⅢa~Ⅲb had a tendency to be greater than in phases Ⅰ and Ⅳ. Conclusions: Resistance to chemotherapy andsensitivity to targeted therapy differ among patients with NSCLC. Differences in gene expression in differentindividuals were also revealed. Only according to personalized detection results can individualized therapeuticregimens be worked out, which is a new direction for oncotherapy.     

胃癌组织中ERCC1、TS和TUBB3与化疗药物体外药敏相关性的实验研究

目的:分析体外胃癌细胞中ERCC1与顺铂、TS与5-氟尿嘧啶及TUBB3与紫杉醇化疗药物敏感性的相关性。方法:选择经外科手术的胃癌患者的新鲜标本48例,每份癌组织标本均分成两份:一份用于原代培养细胞,ATP-TCA法检测体外化疗药物的敏感性;另一份用于免疫组化检测ERCC1、TS和TUBB3蛋白的表达。结果:45例有效的胃癌组织中,ERCC1高表达21例(46.67%),低表达24例(53.33%)。21例ERCC1高表达组敏感例数7例(33.33%),24例低表达组敏感例数17例(70.83%),ERCC1高表达者对顺铂的敏感有效率要明显低于低表达者。TS高表达19例(42.22%),低表达26例(57.78%)。19例TS高表达组敏感例数5例(26.32%),26例低表达组敏感例数18例(69.23%),TS高表达者对5-氟尿嘧啶的敏感有效率要明显低于低表达者。TUBB3高表达22例(48.89%),低表达23例(51.11%)。22例TUBB3高表达组敏感例数5例(22.73%),23例低表达组敏感例数16例(69.57%),TUBB3高表达者对紫杉醇的敏感有效率要明显低于低表达者,以上差异均具有统计学意义(P＜0.05)。结论:体外实验证实ERCC1、TS和TUBB3的表达与化疗药物耐药具有一定的相关性,低表达组的耐药发生率低于高表达组。ERCC1、TS和TUBB3有望成为判断胃癌患者化疗疗效及预后的分子标志物。     

Combined analysis of mRNA expression of ERCC1, BAG-1, BRCA1, RRM1 and TUBB3 to predict prognosis in patients with non-small cell lung cancer who received adjuvant chemotherapy

Background

The aim of this study was to investigate prognostic value of excision repair cross-complementing 1 (ERCC1), BCL2-associated athanogene (BAG-1), the breast and ovarian cancer susceptibility gene 1 (BRCA1), ribonucleotide reductase subunit M1 (RRM1) and class III β-tubulin (TUBB3) in patients with non-small cell lung cancer (NSCLC) who received platinum- based adjuvant chemotherapy.

Methods

Messenger RNA expressions of these genes were examined in 85 tumor tissues and 34 adjacent tissue samples using semi-quantitative RT-PCR. The expressions of these five genes were analyzed in relation to chemotherapy and progression-free survival (PFS) and overall survival (OS). Seventy-four patients were enrolled into chemotherapy.

Results

Patients with ERCC1 or BAG-1 negative expression had a significantly longer PFS (P = 0.001 and P = 0.001) and OS (P = 0.001 and P = 0.001) than those with positive expression. Patients with negative ERCC1 and BAG-1 expression benefited more from platinum regimen (P = 0.001 and P = 0.002). Patients with BRCA1 negative expression might have a longer OS (P = 0.052), but not PFS (P = 0.088) than those with BRCA1 positive expression. A significant relationship was observed between the mRNA expression of ERCC1 and BAG-1 (P = 0.042). In multivariate analysis, ERCC1 and BAG-1 were significantly favorable factors for PFS (P = 0.018 and P = 0.017) and OS (P = 0.027 and P = 0.022).

Conclusions

ERCC1 and BAG-1 are determinants of survival after surgical treatment of NSCLC, and its mRNA expression in tumor tissues could be used to predict the prognosis of NSCLC treated by platinum.     

晚期非小细胞肺癌组织ERCC1和BRCA1表达与铂类化疗敏感性临床研究

目的 吉西他滨联合顺铂是治疗晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)的一线化疗方案,但其有效率仅20％～40％,其较低的化疗有效率与肿瘤耐药相关.研究NSCLC组织中切除修复交叉互补基因1(exeision repair cross-compietionl1,ERCC1)和乳腺癌易感基因1(breast cancer susceptibility gene-1,BRCA1)mRNA的表达及其与铂类化疗敏感性的关系.方法 应用实时荧光定量PCR技术检测2009-03-01-2011-04-16武汉钢铁(集团)公司第二职工医院收集的98例晚期NSCLC患者中ERCC1和BRCA1 mRNA的表达情况,分析其与含铂联合治疗方案化疗耐药性的关系.结果 检测98例患者中ERCC1和BRCA1 mRNA的相对表达量,其中位数分别为0.048和0.011,四分位间距IQR分别为0.0058和0.001.ERCC1 mRNA的高表达率为56.1％(55/98),其表达在不同性别、年龄、组织学类型及临床分期中差异无统计学意义,P＞0.05;BRCA1 mRNA的高表达率为64.3％(63/98),其表达在不同性别、年龄、组织学类型及临床分期中差异无统计学意义,P＞0.05.在ERCC1 mRNA低表达组GP方案化疗客观缓解率为48.8％(21/43),而在高表达组化疗客观缓解率为29.1％(16/55),两组之间比较差异有统计学意义,x2 =4.0,P=0.045;在BRCA1 mRNA低表达组GP化疗客观缓解率为57.1％(20/35),在高表达组GP化疗客观缓解率为28.6％(18/63),差异有统计学意义,x2 =7.74,P=0.005;ERCC1和BRCA1 rnRNA联合低表达组中GP化疗客观缓解率为70.4％(19/27),而其联合高表达组GP化疗客观缓解率仅为21.1％(8/38).结论 ERCC1、BRCA1 mRNA高表达患者在使用含铂化疗方案时疗效差,提示ERCC1和BRCA1 mRNA高表达NSCLC患者对铂类具有耐药性.ERCC1和BRCA1mRNA联合低表达患者在使用含铂方案化疗时有效率更高,提示应用ERCC1和BRCA1 mRNA联合检测时,患者可获得更大的益处.     

Secondary BRCA1 mutations in BRCA1-mutated ovarian carcinomas with platinum resistance

Although ovarian carcinomas with mutated BRCA1 or BRCA2 are sensitive to platinum compounds, such carcinomas eventually develop platinum resistance. Previously, we showed that acquired resistance to cisplatin in BRCA2-mutated tumors can be mediated by secondary intragenic mutations in BRCA2 that restore the wild-type BRCA2 reading frame. Here, we show that secondary mutations of BRCA1 also occur in BRCA1-mutated ovarian cancer with platinum resistance. We evaluated nine recurrent BRCA1-mutated ovarian cancers previously treated with platinum compounds, including five with acquired platinum resistance, one with primary platinum resistance, and three with platinum sensitivity. Four of the six recurrent platinum-resistant tumors had developed secondary genetic changes in BRCA1 that restored the reading frame of the BRCA1 protein, whereas none of the three platinum-sensitive recurrent tumors developed BRCA1 sequence alterations. We immunohistochemically confirmed restored expression of BRCA1 protein in two cases with secondary mutations. Intriguingly, the case with primary platinum resistance showed back mutation of BRCA1 in the primary tumor and showed another secondary mutation in the recurrent tumor. Our results suggest that secondary mutations in BRCA1 can mediate resistance to platinum in BRCA1-mutated ovarian tumors.