乳腺癌患者外周血表皮生长因子受体mRNA与组织中蛋白表达的相关性及其意义

 目的 本研究旨在检测乳腺癌患者外周血EGFRmRNA的表达，评价其作为循环乳腺癌细胞标志的可能性；结合相应肿瘤组织中EGFR蛋白的检测，分析两者的相关性；并结合临床病理资料综合分析，探讨其在乳腺癌患者外周血中表达的意义。方法 以RT-PCR法，分别对40例乳腺癌患者、25例良性乳腺病患者和30例健康人外用静脉血进行EGFRmRNA检测。免疫组化法检测相应肿瘤组织中EGFR蛋白的表达情况。结果 乳腺癌组外周静脉血中EGFRmRNA检出率（32．5％）与健康人组（0）和良性乳腺病患者组（0）间差异均存在统计学意义（P〈0．01）。免疫组化法检测到乳腺癌患者相应肿瘤组织中有25例（62．5％）EGFR蛋白阳性，组织中EGFR蛋白表达级别高的患者其相应外周血EG—FRmRNA的检出率也高。EGFRmRNA阳性表达与临床分期、组织学分级及ER闰存在明显的相关性，而与病理类型、肿瘤大小、腋窝淋巴结转移、PR和HER-2间无明显相关性。结论 RT-PCR法检测乳腺癌患者外周血EGFRmRNA有望成为检测循环乳腺癌细胞的有效方法。结合临床病理资料分析，RT—PCR法检测外周血EGFRmRNA有助于判断乳腺癌的预后，有益于乳腺癌微转移的早期诊断、治疗方案的选择及治疗疗效的监测。     

应用RT-PCR检测乳腺癌患者外周血CEA mRNA和CK-19 mRNA的研究及意义

目的：检测乳腺癌患者外周血中CEA mRNA和CK-19 mRNA的表达，建立取材于外周血早期诊断乳腺癌微转移的方法。方法：65例乳腺癌和37例乳腺良性疾病患者的外周血，分离其有核细胞后进行细胞总RNA的抽提，运用果式RT-PCR技术进行CEA mRNA和CK-19 mRNA的检测。结果：以     

应用RT-PCR检测乳腺癌患者外周血CK-19mRNA的临床意义

目的:探索取材于乳腺癌患者外周血诊断微转移的方法。方法:66例乳腺癌和37例乳腺良性疾病患者的外周血,分离其有核细胞后进行细胞总RNA的抽提,运用RT-PCR技术进行CK-19 mRNA的检测。结果:以RT-PCR终产物出现460bp条带定为阳性。66例乳腺癌患者CK-19mRNA有24例表达,总阳性率为36.36％。37例乳腺良性疾病者CK-19mRNA无一例表达。两组有统计学意义(x2=17.54,P<0.001)。结论:RT-PCR检测CK-19 mRNA可以用于临床诊断乳腺癌患者外周血中微转移。     

定量RT-PCR检测乳腺癌患者外周血中 CK19 mRNA表达的意义

目的:建立实时荧光定量逆转录-多聚酶链反应法(RT-PCR)检测乳腺癌患者外周血细胞角蛋白19(CK19)mRNA表达的状况,并探讨其临床意义.方法:70例经病理诊断的乳腺癌患者和30名健康对照者,以实时荧光定量RT-PCR检测其外周血CK19 mRNA的表达.结果:70例乳癌患者外周血CK19 mRNA表达的阳性率为32.9%(23/70),30例对照者的阳性率为3.3%(1/30),经统计学分析,两者差异具有显著性差异(P=0.002).在不同分期的乳腺癌患者中均存在不同程度CK19 mRNA的表达,Ⅰ、Ⅱ、Ⅲ期患者外周血CK19 mRNA阳性率随分期增加而增高.结论:实时荧光定量RT-PCR方法可检测出各期乳腺癌患者外周血中CK19 mRNA的表达,具有较高的敏感性和特异性,提示循环肿瘤细胞可能存在于乳腺癌发展的各个阶段.     

乳腺癌患者外周血中CEA mRNA表达的检测及临床意义

目的探讨RT—PCR法检测乳腺癌患者外周血中的CEA mRNA表达的情况及在乳腺癌外周血微转移诊断中的应用。方法采用RT—PCR法,检测46例乳腺癌患者外周血中CEA mRNA表达情况,以良性乳腺疾病患者及健康志愿者外周血标本作为对照。结果46例乳腺癌患者外周血中CEA mRNA阳性率为32．6％。对照组中CEA mRNA表达均为阴性。CEA mRNA的阳性表达与患者的TNM分期、腋淋巴结状况差异有统计学意义（P〈0．05）,而与患者的年龄、原发肿瘤大小、病理类型以及激素受体状况等差异均无统计学意义（P〉0．05）。结论CEA mRNA可作为标志物来检测乳腺癌患者外周血中微转移,有助于指导临床治疗及判断预后。     

无假基因干扰的CK19 RT-PCR方法检测乳腺癌外周血微转移

目的:以CK19为基因标志检测乳腺癌患者外周血中播散的肿瘤细胞,并对其临床意义进行评价。方法:采用优化的无假基因干扰的RT-PCR方法检测乳腺癌患者外周血有核细胞的CK19mRNA表达。结果:1)方法可靠性验证:检测30例正常人外周血CK19mRNA表达,结果无假阳性;将乳腺癌细胞系MCF-7以1∶107与正常外周血有核细胞混合,检测CK19mRNA表达,结果无假阴性。2)检测乳腺癌41例,外周血肿瘤细胞阳性检出率26.8%(11/41);乳腺癌患者外周血肿瘤细胞阳性率在患者不同年龄、肿瘤大小、临床分期、组织学分级、ER和PR状态的各组间差异无显著性(P>0.05);淋巴结转移阳性病例外周血肿瘤细胞检出率(38.1%)明显高于淋巴结转移阴性组(15.0%),虽然统计学差异不具有显著性(P=0.10),但提示淋巴结转移阳性的患者肿瘤细胞血行播散的危险性有增加的趋势。结论:以CK19为基因标志采用RT-PCR方法检测乳腺癌患者外周血中的肿瘤细胞可能是乳腺癌有意义的预后指标,可以为治疗方案的选择、治疗疗效的评价及外周血干细胞移植联合大剂量化疗适应证筛选提供重要依据。     

乳腺癌组织中表皮生长因子受体（EGFR）的研究

用￣（１２５）Ｉ标记的表皮生长因子（￣（１２５）Ｉ－ＥＧＦ）作为配基，用标准的ＥＧＦ作为竞争剂，采用单点饱和分析法测定了３６例乳腺癌组织中的ＥＧＦＲ含量，范围在１～１２２ｆｍｏｌ／ｍｇ膜蛋白之间，以１０ｆｍｏｌ／ｍｇ膜蛋白以上为阳性，乳腺癌阳性率为３８％。按乳腺癌临床分期的结果表明，临床分期越高，阳性率也越高。有淋巴结转移者较无淋巴结转移的病人阳性率有增高趋势。证明表皮生长因子受体与乳腺癌的恶性程度有关，但癌各组织学类型之间阳性率未见差别。     

早期乳腺癌患者外周血CK19 mRNA阳性循环细胞检测及其临床意义

 目的　探讨早期乳腺癌患者外周血CK19 mRNA阳性细胞检测的临床意义及其预后价值。方法　应用RT-PCR检测50例早期乳腺癌患者、24例良性乳腺病变患者及20名健康体检志愿者外周血有核细胞CK19 mRNA的表达,并根据基因标志物结果随访。结果　早期乳腺癌患者术后辅助治疗前外周血有核细胞CK19 mRNA阳性率40.0 %（20／50）,与良性乳腺疾病患者[12.5 %（3／24）]及健康体检志愿者[5 %（1／20）]比较,差异均有统计学意义（P＝0.018,P＝0.004）;外周血有核细胞CK19 mRNA阳性率与患者年龄、月经状况、TNM分期、肿瘤大小、淋巴结转移状况、病理分化、激素受体状况、Her-2、血清CA153、血清CEA水平异常升高无关（P＞0.05）;20例外周血有核细胞CK19 mRNA阳性的患者中有11例随访期出现转移复发（P＝0.002）,中位无瘤生存期明显缩短（P＝0.007）。结论　CK19 mRNA可以作为循环肿瘤细胞基因标志物。检测CK19 mRNA阳性循环肿瘤细胞,可能作为早期乳腺癌术后判断复发转移的辅助指标。     

表皮生长因子受体反义重组腺病毒联合放射线对乳腺癌细胞的作用

目的探讨表皮生长因子受体(EGFR)反义重组腺病毒联合放射线对乳腺癌细胞的作用。方法人EGFR的cDNA片段被反向亚克隆入E1／E3缺失的5型腺病毒载体中而得到AdE5，EGFR反义RNA的表达由CMV启动子控制。进而研究其联合γ射线对人乳腺癌细胞株MDA-MB-231成克隆能力及细胞周期分布的影响。结果MDA-MB-231细胞被AdE5感染后，EGFR蛋白质的表达被强烈地抑制。AdE5感染后以γ射线照射细胞，成克隆能力被抑制。且这种作用与病毒量和照射剂量相关联。进而流式细胞仪分析显示，300pfu／细胞的AdE5感染可使MDA-MB-231细胞摆脱对放射线抗拒的G0 G1期，进入对放射线敏感的G2 M期，从而使AdE5联合放射线表现为协同效应。结论以腺病毒载体介导的EGFR反义RNA转导可有效地用于针对EGFR的反义治疗策略，提高放射线对乳腺癌细胞杀灭作用。     

RT-PCR检测K19 mRNA诊断直肠癌区域淋巴结微转移

目的：探讨RT—PCR检测K19 mRNA诊断直肠癌区域淋巴结微转移的价值及其应用前景。方法：通过病理组织学及RT—PCR扩增K19 mRNA对19例直肠癌患者的癌组织标本和51个淋巴结进行检测。结果：19例直肠癌患者癌组织均有K19 mRNA表达；51个淋巴结中组织学阳性16个（31．4％），而RT—PCR阳性21个（41．2％）。RT—PCR扩增K19 mRNA及内参照β—actin后，所有癌组织及转移淋巴结显示461bp和221bp的扩增片断，而非肿瘤患者的8个淋巴结仅显示221bp扩增片断。结论：RT—PCR扩增K19 mRNA检测直肠癌区域淋巴结微转移是一种敏感、特异的检测方法，优于常规病理组织学方法。     

RT—PCR检测K19 mRNA诊断直肠癌区域淋巴结微转移

目的：探讨RT—PCR检测K19 mRNA诊断直肠癌区域淋巴结微转移的价值及其应用前景。方法：通过病理组织学及RT—PCR扩增K19 mRNA对19例直肠癌患者的癌组织标本和51个淋巴结进行检测。结果：19例直肠癌患者癌组织均有K19 mRNA表达；51个淋巴结中组织学阳性16个（31．4％），而RT—PCR阳性21个（41．2％）。RT—PCR扩增K19 mRNA及内参照β—actin后，所有癌组织及转移淋巴结显示461bp和221bp的扩增片断，而非肿瘤患者的8个淋巴结仅显示221bp扩增片断。结论：RT—PCR扩增K19 mRNA检测直肠癌区域淋巴结微转移是一种敏感、特异的检测方法，优于常规病理组织学方法。     

hMAM mRNA的定量检测对乳腺癌外周血微转移的诊断价值

目的:建立SYBR Green I实时定量逆转录PCR(Real-Ti me QuntitativeRT-PCR,Q-RT-PCR)技术定量检测人乳腺珠蛋白(human mammaglobin,hMAM)mRNA的技术,并应用于检测外周血hMAM mRNA,探讨其诊断乳腺癌患者外周血微转移的可行性及意义。方法:应用自行建立的SYBR Green I Q-RT-PCR检测技术,检测61例乳腺癌、33例乳腺良性肿瘤患者以及11例健康志愿者外周血hMAM mRNA的表达量,分析其表达水平与乳腺癌临床病理特征及ER、PR及HER-2表达的关系。结果:61例乳腺癌患者外周血中10例检出hMAM mRNA表达,其癌组织HER-2阳性者占8例,HER-2阴性组未见hMAM mRNA阳性表达。HER-2阳、阴性组间外周血hMAM mRNA表达,差异具有统计学意义,P=0.028;与其他临床、病理特征无相关性,P>0.05。HER-2阳性患者外周血中hMAM mRNA表达量范围1.98×102~6.21×103,中位数4.0×102,在临床、病理特征间差异无统计学意义,P>0.05。乳腺良性肿瘤及健康志愿者外周血hMAM mRNA均阴性。结论:SYBR GreenI Q-RT-PCR技术可定量检出乳腺癌患者外周血hMAM mRNA;乳腺癌患者外周血hMAM mRNA表达与其乳腺癌组织HER-2表达相关。定量检测hMAM mRNA可作为乳腺癌外周血微转移的重要方法。     

Prognostic value of epidermal growth factor receptor in node-positive breast cancer

Summary The prognostic significance of EGFR (epidermal growth factor receptor) was studied in a cohort of 68 node-positive patients with breast cancer, who entered a controlled protocol of adjuvant therapy between February 1980 and June 1984. EGFR radioligand binding assay was carried out on frozen stored samples. Twenty five (37%) of 68 primary sites and 9 (41%) of 19 lymph node metastases assayed were EGFR-positive with a cut off value of 5 fmol/mg membrane protein; there is no statistical difference between the two distributions. EGFR was significantly correlated to ER and histological grade. EGFR-positive tumors and high levels of EGFR were mainly found in the ER-negative group of tumors (p = 0.008) and in histological grade III (p = 0.007). Fifty five patients could be followed for 40 to 92 months. EGFR was an independent prognostic factor for survival after 40 months (p = 0.05). EGFR⁺/ER^– patients had the lowest survival probability, but statistical significance was not reached (p = 0.06). The EGFR phenotype appeared as a prognostic parameter in node-positive patients, individualizing subgroups of patients with different early outcome, with potential therapeutic implication especially in the group of ER-negative patients. These results emphasize the need for a standardized assay methodology and for further clinical studies, particularly in protocols where adjuvant hormonal therapy is prescribed on the basis of steroid hormone receptor status, in order to assess the respective prognostic worth of EGFR and ER (or PR).     

实时荧光定量PCR检测非小细胞肺癌患者外周血和肿瘤组织表皮生长因子受体突变的研究

目的应用实时荧光定量一PCR(FQ-PCR)对非小细胞肺癌(NSCLC)患者外周血和肿瘤组织表皮生长因子受体(EGFR)突变进行检测,并分析该方法对肿瘤组织和外周血EGFR突变的一致性.方法采用FQ-PCR检测112例NSCLC外周血和87例NSCLC组织EGFR突变,对其中45例外周血和肿瘤组织进行配对检测,并研究分...     

人表皮生长因子受体-2及雌激素受体与乳腺癌的关系

乳腺癌与人表皮生长因子受体-2及雌激素受体关系密切,这两种受体也是乳腺癌的分类标准和治疗靶点。在大多数乳腺癌患者中,人表皮生长因子受体-2信号途径和雌激素受体信号途径参与了细胞的增生存活过程。而且在乳腺癌病例中,人表皮生长因子受体-2和雌激素受体呈现出一定程度的负相关。这说明这两种受体活化后有一些联系。本文简要综述了人表皮生长因子受体-2和雌激素受体的联系以及这种联系在乳腺癌治疗中的意义。     

Expression of epidermal growth factor receptor mRNA and protein in primary breast carcinomas

The expression of epidermal growth factor receptor (EGFR) mRNA and protein has been determined in a group of breast carcinomas and compared to oestrogen and progesterone receptor (ER, PgR) status, as well as pathological features. In situ hybridization using a digoxigenin-labelled oligonucleotide probe was applied to formalin-fixed paraffin-embedded sections, and immunohistochemistry was used to determine EGFR protein.EGFR mRNA was detected in 66% of carcinomas with a third having labelling similar to normal breast tissue, 22% heterogeneous weak to strong labelling, and 11% strong labelling. EGFR protein was detected in 36% and these tumours had a strong correlation to lack of ER and high histological grade. The presence of EGFR protein was strongly correlated with more intense labelling for EGFR mRNA (p < 0.0001). This contrasted with normal breast in which both EGFR protein and mRNA were present with varying degrees in both tumours and a normal breast control. The ER-/PgR- carcinomas showed the full range of EGFR mRNA labelling. It is postulated that oestrogen or oestrogen regulated proteins are involved in regulation of EGFR mRNA and protein. In a proportion of tumours lacking steroid receptors regulation is lost, leading to EGFR overexpression.     

Role of epidermal growth factor receptor expression in primary breast cancer: results of a biochemical study and an immunocytochemical study

Summary We have conducted two series of studies, a biochemical study and an immunocytochemical study, to investigate the role of epidermal growth factor receptor (EGFR) expression in primary breast cancer patients. In the biochemical study, a consecutive 115 patients were included and EGFR was measured by a competitive binding assay with multipoint Scatchard analysis. In the immunocytochemical study comprising 126 patients, EGFR status was determined by immunostaining with anti-EGFR antibody EGFR1. Several agreements were found from these two studies. EGFR status was inversely correlated with estrogen receptor (ER) status. No significant correlation was found between EGFR status and tumor size, nodal metastases, or the expression of c-erbB-2 protein. Ki-67 immunoreactivity, a cellular proliferation marker, was enhanced in EGFR positive tumors over EGFR negative tumors, suggesting a linkage of EGFR expression to cellular proliferative activity. Post-operative follow up showed that relapse-free survival for EGFR positive patients was significantly worse than that for EGFR negative patients, particularly in node-positive patients. Multivariate analysis demonstrated a significance of EGFR status as an independent prognostic indicator in primary breast cancer. The group expressing EGFR and c-erbB-2 protein indicated a particularly high risk for relapse.