Epidermal growth factor receptor-targeted immune magnetic liposomes capture circulating colorectal tumor cells efficiently

AIM To compare the capacity of newly developed epidermal growth factor receptor(EGFR)-targeted immune magnetic liposomes(EILs) vs epithelial cell adhesion molecule(Ep CAM) immunomagnetic beads to capture colorectal circulating tumor cells(CTCs).METHODS EILs were prepared using a two-step method, and the magnetic and surface characteristics were confirmed. The efficiency of capturing colorectal CTCs as well as the specificity were compared between EILs and Ep CAM magnetic beads. RESULTS The obtained EILs had a lipid nanoparticle structure similar to cell membrane. Improved binding with cancer cells was seen in EILs compared with the method of coupling nano/microspheres with antibody. The binding increased as the contact time extended. Compared with Ep CAM immunomagnetic beads, EILs captured more CTCs in peripheral blood from colorectal cancer patients. The captured cells showed consistency with clinical diagnosis and pathology. Mutation analysis showed same results between captured CTCs and cancer tissues. CONCLUSION EGFR antibody-coated magnetic liposomes show high efficiency and specificity in capturing colorectal CTCs.     

Epidermal growth factor receptor and cyclin D1 are independently amplified and overexpressed in esophageal squamous cell carcinoma

Purpose To assess the status of EGFR, HER-2, and CCND1 at the gene and protein levels in esophageal squamous cell carcinoma.Methods Dual-color FISH assays were performed using DNA probes for EGFR/CEP 7, HER-2/CEP 17, and CCND1/CEP 11. The respective proteins, furthermore, was assessed in IHC assays and correlated with patient and tumor characteristics.Results From 55 ESCCs, 8 (15%) tumors showed gene amplification and 20 (36%) had gene overrepresentation (balanced gene and chromosome 7 polysomy) for EGFR. High-level protein expression was frequent (49%), positively correlated with gene copy numbers (kappa=0.4), and associated with well-differentiated histology (p=0.02). For HER-2, gene amplification was detected in a single tumor (2%) and protein overexpression was rare (9%). CCND1 gene was amplified in 23 (42%) tumors; likewise, CCND1 protein overexpression was common (58%) and prevailed in gene overrepresentation or amplification. Only 1 patient showed gene amplification for both EGFR and CCND1. Survival was not associated with EGFR or CCND1 gene/protein status, whereas negative patients for HER-2 protein had a better survival than positive patients (p=0.04).Conclusions Frequent overexpression and gene amplification of EGFR and CCND1 make these molecules and their pathways potential therapeutic targets for ESCC. In addition, EGFR and CCND1 appeared to be independently altered suggesting alternative mechanisms for pathway activation. Therapeutic agents targeting these molecules are urged to be tested in clinical trials and comprehensive biological analyses should be included to properly interpret the outcome.     

In vitro effect of radiation, antibody to epidermal growth factor receptor and Docetaxel in human head and neck squamous carcinoma cells with mutant P53 and over-expressed EGFR

Purpose  Radiotherapy is the most frequently used and cheapest treatment both for curative and palliative purposes in HNSCC. Despite advances in technology and intensive treatments with radiation, only half of the patients are cured. New therapeutic approaches focusing on the molecular mechanism that mediate tumour cell growth or cell death in combination with radiotherapy have been suggested. The effects of radiation, antibody to EGFR and Docetaxel as single treatment or in combinations on HNSCC cells were investigated. Methods  The established HNSCC cells with mutant (mt) P53 and over-expressed normal EGFR was used as the in vitro model. Gene expression profile, cell cycle progression and cell death were used as the indication of treatment outcome. Results  With c-DNA microarray of well-characterised functional genes, massive changes in the genes expression of HNSCC were detected. The alterations of gene expression profiles do not have any correlation neither on tumour cell growth nor cell death. HNSCC cells with mt P53 and over-expressed normal EGFR did not response to radiation, anti-EGFR monoclonal antibody and their combination therapy. Effective treatment could be obtained from single therapy with Docetaxel. No additive effects on cell cycle arrest or cell death were seen in the combination of Docetaxel to anti-EGFR antibody, radiation or anti-EGFR antibody + radiation. Conclusions  The c-DNA microarray analysis does not indicate any specific target or treatment effects of HNSCC with mt P53 and over-expressed normal EGFR. Single therapy, target at microtubules might be the most suitable treatment modulation in this tumour type.     

气管腺样囊性癌表皮生长因子受体基因突变的检测

目的了解气管腺样囊性癌肿瘤组织表皮生长因子受体（EGFR）18、19、20、21位点基因突变情况,为气管腺样囊性癌的分子靶向治疗奠定基础。方法将自2004年至2013年在煤炭总医院经气管镜下取出的气管腺样囊性癌共36例蜡块标本,提取肿瘤细胞DNA,采用ARMS法进行EGFRl8、19、20、21位点基因突变检测。采用Fisher精确概率法比较两组之间阳性率的差别。P〈0．05为差异有统计学意义。结果36例气管腺样囊性癌蜡块包埋标本中,EGFR基因突变阳性率为31％（11／36）。14％（5／36）存在双突变（19外显子缺失突变及21外显子突变）,0％（0／36）出现EGFR基因20外显子突变。临床分期为Ⅳ期的标本EGFR基因突变率为63％（5／8）,临床分期为Ⅱ～Ⅲ期的标本中EGFR基因突变率为21％（6／28）,两组间有明显差异（P〈0．05）。结论气管腺样囊性癌EGFR基因突变率介于肺腺癌与鳞癌之间。气管腺样囊性癌EGFR基因突变在已有血行转移的患者中阳性率明显高于无血行转移者,晚期气管腺样囊性癌患者可能从EGFR-酪氨酸激酶抑制剂治疗中获益。     

表皮生长因子与胃肠道疾病

表皮生长因子(EGF)是重要的促细胞生长因子之一,消化道中EGF的含量远远高于血液循环中的浓度,本文就EGF的结构、性质、生理作用、合成部位、分布情况以及与良恶性胃病的关系和对胃肠道疾病的治疗作用做一综述。     

非小细胞肺癌患者血清血管内皮生长因子、血小板衍生生长因子和表皮生长因子受体测定的临床意义

目的 探讨血清血管内皮生长因子(vascular endothelial growth factor,VEGF)、血小板衍生生长因子(platelet-derived growth factor,PDGF)和表皮生长因子受体(epidermal growth factor receptor,EGFR)测定在非小细胞肺癌(non-small cell lung cancer,NSCLC)诊断和预后判定中的意义.方法 采用双抗体夹心ABC-ELISA法测定31例NSCLC患者及30名健康者血清VEGF、PDGF和EGFR的含量.结果 NSCLC患者血清VEGF、PDGF和EGFR测定值均高于健康对照组(P值均＜0.01).血清VEGF、PDGF和EGFR测定值与NSCLC病理分型无关(P值均＞0.05),与远处转移有关,远处转移组的测定值高于未转移组(P＜0.05～0.01).NSCLC患者血清VEGF与PDGF测定值之间呈显著正相关(r=0.641,P＜0.01),血清VEGF和EGFR测定值呈正相关(r=0.369,P＜0.05).结论 检测血清VEGF、PDGF和EGFR水平对NSCLC的诊断和预后判定具有一定价值.     

Epidermal growth factor receptor gene mutation,amplification and protein expression in malignant pleural mesothelioma

Epidermal growth factor receptor (EGFR) is overexpressed in a variety of epithelial malignancies. In lung cancer cases, EGFR gene mutation at the kinase domain and EGFR gene amplification are reported to be predictors of the response to EGFR tyrosine kinase inhibitors. In malignant pleural mesothelioma (MPM), the role of EGFR is less clear. We studied EGFR gene mutation, amplification and protein expression in 25 Japanese patients with MPM. None had previously reported EGFR mutations detected by the TaqMan PCR assay. Using immunohistochemistry, 8/25 (32%) cases were positive for the EGFR protein. The cases of sarcomatous type and desmoplastic type were all negative. Fluorescence in situ hybridization analysis revealed three low polysomy cases and one high polysomy case. The low polysomy cases included one biphasic type and two epithelial types, and the high polysomy case was epithelial type. These four cases expressed EGFR protein. In MPM, EGFR seems to play a role in a limited subset of patients. To identify possible candidates for EGFR tyrosine kinase in inhibitor therapy, the information on the EGFR gene status may be valuable.     

胃癌患者检测表皮生长因子及受体的意义

目的:探讨表皮生长因子(EGF)及其受体(EGFR)的表达与胃癌发生及胃癌生物学行为的关系。方法:采用免疫组化S-P法对50例胃癌进行研究。结果:EGF和EGFR在早期胃癌中的阳性率均为20％(2/10),在进展期胃癌的阳性率分别为62.5％(25/40)和60％(24/40),进展期胃癌EGF和EGFR的阳性率均显著高于早期胃癌(P<0.05)。有转移组的EGF及EGFR阳性率高于无转移组(P<0.05)。EGF及EGFR的表达与胃癌的组织学类型有关。结论:EGF及EGFR阳性的肿瘤可能具有更强的浸润与转移能力,检测EGF和EGFR有助于判断胃癌预后。     

Epidermal growth factor receptor directed therapy in head and neck cancer

Squamous cell head and neck cancer (SCCHN) is the seventh most common cause of cancer death worldwide and its incidence is rising rapidly in developing countries. Despite recent advances in managing locally advanced SCCHN, patients with recurrent and metastatic SCCHN have a poor prognosis and little progress has been made its management. Epidermal growth factor receptor (EGFR) has been implicated in the pathogenesis of SCCHN and is a marker of poor prognosis. Recent advances in targeted therapeutics against EGFR are being investigated clinically. In this article, we review the different modalities utilized to inhibit EGFR signaling in SCCHN, including small molecule tyrosine kinase inhibitors, monoclonal antibodies, anti-sense therapy and immunotoxin conjugates. Monotherapy with EGFR inhibitors has demonstrated response rates between 5 and 15% in advanced SCCHN. However, combining EGFR inhibitors with cytotoxic chemotherapy or radiation therapy appears to augment response rates and survival. With the foundation for the use of EGFR inhibitors laid in these studies, future studies will need to optimize the delivery of these agents in combination with conventional therapies.     

表皮生长因子治疗糖尿病足溃疡的研究进展

糖尿病足是糖尿病常见的慢性并发症之一,15%的糖尿病患者可能发生足部溃疡。溃疡创面局部生长因子及受体活性下降和数量的绝对或相对缺乏是其难以愈合的病理生理基础。表皮生长因子(EGF)通过促进细胞迁移、增殖及细胞外基质合成等参与溃疡创面愈合。外源性EGF作为一种新的治疗手段,局部应用于糖尿病足溃疡取得显著效果。通过组织工程学技术,EGF释药方式不断改善,并可与其他生长因子联合应用,具有广泛前景。     

表皮生长因子受体及增殖细胞核抗原在胃癌中的表达及意义

为探讨表皮生长因子受体 ( EGFR)及增殖细胞核抗原 ( PCNA)在胃癌中的表达 ,应用免疫组织化学技术 ,对胃癌和胃良性肿瘤组织中 EGFR和 PCNA分别进行定性和半定量检测 ,分析其与临床病理参数的关系。结果显示 1在胃良性肿瘤组织中 EGFR的表达率为 2 3 .0 8% ,PCNA L I为 2 7.80± 12 .2 0 ,明显低于胃癌组织中的表达率 ( 49.0 9% ,PCNA L I5 2 .73± 12 .2 5 ,P<0 .0 1)。2胃癌组织中 EGFR和 PCNA的表达与患者性别、年龄、病变部位、分型、组织学类型及分化程度无关 ( P均 >0 .0 5 ) ,而与临床分期、浸润深度及淋巴结转移情况相关 ( P <0 .0 5 ) ,表明联合检测 EGFR和 PCNA可作为判断胃癌恶性程度、预测淋巴结转移、制定合理治疗方案及评估预后的重要指标     

非小细胞肺癌表皮生长因子受体基因突变与分子靶向治疗

肺癌分子靶向治疗近年来取得较大进展.特别是针对表皮生长因子受体(EGFR)分子靶向药物表现出确定的临床效果.临床应用表明,EGFR基因酪氨酸激酶域体细胞突变与非小细胞肺癌患者对酪氨酸激酶抑制剂吉非替尼的敏感性相关.本文就EGFR生物学特点、EGFR在肺癌中的表达、EGFR基因突变率、EGFR基因突变与EGFR酪氨酸激酶抑制剂疗效的关系作一综述.     

转化生长因子α对小鼠气道平滑肌细胞的促增殖作用

目的 探讨转化生长因子α(transforming growth factog alphg,TGF-α)对小鼠气道平滑肌细胞(airway smooth muscle cells,ASMC)促增殖作用及其机制.方法 用四唑盐(MTT)比色法和3H-TdR掺人法测定加入TGF-α后小鼠ASMC的增殖情况.本实验采用3H-TdR掺入法和MTT比色法观察选择性表皮生长因子受体(epidermal growth factor receptor,EGFR)酪氨酸激酶抑制剂(AG1478)、EGFR中和抗体225(225mAb)、MEK抑制剂(U0126)、PI-3K抑制剂(Wonmannin)对加入TGF-α后促ASMC增殖的影响.通过Western Blot方法测定加入TGF-α及加用AG1478、225mAb后ASMC磷酸化EGFR蛋白表达.结果 用MTT比色法和3H-TdR掺入法测定ASMC培养液中加入TGF-α后ASMC增殖情况比对照组明显增加.加入AG1478、225mAb、U0126、Wortmannin可抑制TGF-α促ASMC增殖作用(P＜0.01).经Western Blot检测,TGF-α引起ASMC磷酸化EGFR蛋白表达增高.AG1478、225mAb抑制TGF-α所致ASMC磷酸化EGFR蛋白表达的增加(P＜0.01).结论 TGF-α激活EGFR为磷酸化EGFR,从而通过:①ras-raf-MEK-erk/MAPK途径;②PI3K-PKC-IKK途径;促进体外培养的小鼠ASMC的增殖.     

Growth requirements,growth factor responsiveness,and growth factor secretion of three human embryonal carcinoma cell lines

Summary The in vitro growth requirements of three human embryonal carcinoma cell lines (H 12.7, 2102 EP, 1428 A) were investigated. The basal medium DME/F12 supplemented with insulin, transferrin, and low-density and high-density lipoproteins was sufficient to support substantial multiplication of all three lines. The most efficient attachment factor was either fibronectin (for 2102 EP and 1428 A) or collagen type I (H 12.7). In a serum-free system the influence of epidermal growth factor (EGF), insulin-like growth factor I, multiplication stimulating activity (MSA), a platelet extract, and the glucocorticoids dexamethasone and hydrocortisone, as determined by the DNA synthesis rate of the cells, was generally minimal. However, the DNA synthesis rate of cell lines H 12.7 and 2102 EP was increased by MSA, and the line with the highest potential to differentiate (H 12.7) was stimulated by EGF. All three cell lines secreted growth factors in a heterologous stimulation assay. Insulin-like growth factors I and II were not part of the growth promoting activity. The inhibitory effect of a monoclonal anti-EGF antibody on the ³H-thymidine incorporation of cell line 2102 EP might indicate autocrine secretion of EGF or an EGF-like factor by this cell line.     

表皮生长因子受体抑制剂在非小细胞肺癌治疗中的研究进展

化疗一直是晚期非小细胞肺癌(NSCLC)的主要治疗手段之一,但是治疗的有效率仅为30％左右,且具有较大的不良反应,耐药和血液学的不良反应是患者预后不良的主要原因之一.随着分子生物标记如表皮生长因子受体(EGFR)等的发现和相应靶向药物的研发与批准应用,肺癌的治疗正在发生重大变化.大量研究证实EGFR抑制剂靶向治疗能改善NSCLC患者的疾病无进展期和生活质量,而正确的选择合适的患者是靶向治疗成功的关键.本文将对EGFR的组成及活化、检测方法、EGFR靶向药物在NSCLC治疗中的作用以及获得性耐药患者的治疗进展进行综述.     

非小细胞肺癌EGFR基因突变检测与临床意义

目的 探索铜陵地区非小细胞肺癌表皮生长因子受体(EGFR)基因突变与其临床病理特征的关系.方法 用蝎形探针扩增阻滞突变系统对非小细胞肺癌患者石蜡包埋组织、冷冻组织进行EGFR基因18～21外显子突变的检测,分析EGFR基因的突变的分布特征及其与临床病理特征的相关性.结果 42份非小细胞肺癌标本中24份检测出EGFR基因突变,总检出率57.14％.突变类型主要包括L858R点突变(41.67％)、19-del E746-A450缺失突变(41.67％)、L858R+ 19-del(12.50％)、G719X点突变(4.17％).未检测出第20外显子T790M点突变.女性患者突变率(75.00％)较男性患者突变率(40.91％)高,二者差异有统计学意义(P=0.026);吸烟＜400年支突变率为70.37％,较吸烟≥400年支突变率(33.33％)高,差异有统计学意义(P=0.02);肿瘤病理学类型为腺癌突变率为60.61％(20/33),而鳞癌突变率为40.00％ (2/5),二者差异无统计学意义(P=0.346).年龄＜60岁患者突变率为64.00％,较≥60岁患者突变率(47.06％)高,差异无统计学意义(P=0.276).不同TNM分期之间突变率差异无统计学意义(P=0.9).结论 EGFR基因突变多见于女性、不吸烟或少吸烟非小细胞肺癌患者.     

吉非替尼对肺腺癌细胞增殖和表皮生长因子受体表达的影响

目的 探讨表皮生长因子受体(EGFR)抑制剂吉非替尼对肺腺癌A549细胞株增殖和EGFR表达的影响.方法 不同浓度吉非替尼干预肺癌A549细胞24、48、72 h后,倒置相差显微镜观察药物干预后细胞形态学变化;四甲基偶氮唑盐(MTT)比色法测定细胞抑制率;钙依赖性磷脂结合蛋白(AnnexinⅤ)/碘化丙啶(PI)法和HoechsT_33258染色法检测细胞凋亡;流式细胞仪检测药物作用周期;免疫荧光检测EGFR蛋白表达情况;实时荧光定量PCR检测EGFR mRNA的表达情况.结果 各干预组细胞出现空泡和颗粒,细胞碎片增多,且随吉非替尼剂量增加和干预时间的延长日益明显.吉非替尼明显抑制了A549细胞的生长,呈时间、剂量依赖性.吉非替尼组细胞凋亡率明显高于正常对照组(P<0.01),S期细胞比例明显减少(P<0.01),G0/G1期细胞比例明显增加(P<0.01),EGFR mRNA和蛋白表达明显减弱(P<0.01).结论 吉非替尼显著抑制A549细胞生长,可能机制是促进凋亡、增强G0~G1期阻滞和进一步下调活化的EGFR mRNA和蛋白的表达.     

胃癌组织中EGFR和COX-2表达的意义及其相关性

目的:探讨表皮生长因子受体(EGFR)和环氧化酶-2(COX-2)的表达与胃癌侵袭、转移因素的关系以及EGFR与COX-2在胃癌组织中表达的相关性.方法:用免疫组化的方法检测61例胃癌组织和相应的20例癌旁组织石蜡切片中EGFR、COX-2表达情况:用Western blot方法检测10例胃癌组织和相应的癌旁组织中EGFR、COX-2蛋白的表达.结果:免疫组化检测胃癌组织中EGFR及COX-2的阳性表达率分别为36.07%、59.02%均明显高于癌旁组织的0%、25%(χ~2=9.903,P<0.01;χ~2=6.972,P<0.01);COX-2表达与浸润深度、淋巴结转移、TNM分期和病理分化程度有关(χ~2=6.333,P<0.05;χ~2=5.588,P<0.05;χ~2=8.826,P<0.01;χ~2=5.653,P<0.05).EGFR表达与淋巴结转移和TNM分期有关(χ~2 =10.648,P<0.01;χ~2=4.150,P<0.05).EGFR与COX-2表达呈明显相关(r=0.316,P<0.05).Western blot方法检测胃癌组织中EGFR及COX-2的蛋白表达高于癌旁组织(35.89±12.50 vs 15.14±2.15,P<0.01;51.29±23.42 vs 27.65±7.42,P<0.05).结论:EGFR、COX-2在胃癌组织中高表达,EGFR、COX-2的表达与胃癌的侵袭、转移密切有关,COX-2表达与EGFR显著相关.