Immunohistochemical study of epidermal growth factor receptor in adenoid cystic carcinoma of salivary gland origin

BACKGROUND: Epidermal growth factor (EGF) and its receptor (EGFR) are involved in the development of salivary gland tumors. Recently, treatment modalities for EGFR inhibition have shown an enhanced clinical response in carcinomas of different locations. Adenoid cystic carcinoma (ACC) of salivary gland origin is a malignant tumor with a poor long-term outcome. If salivary gland ACC does exhibit EGFR, then immunotherapy could have a major impact on improving its prognosis. METHODS: The study consisted of 34 samples of formalin-fixed, paraffin-embedded specimens of salivary gland ACC. Specimens were stained with a mouse antihuman monoclonal antibody for immunohistochemical detection of EGFR. Overlying oral mucosa and adjacent normal salivary ducts served as internal controls. Both membrane and cytoplasmic staining were evaluated. Staining score was calculated by multiplying the percentage of positively stained tumor cells by the intensity of the staining. The highest score for a given tumor was equal to 2. RESULTS: In the final analysis, 27 of the 34 specimens were included; 7 were excluded, because the internal control did not reveal any staining. Of these 27 specimens, 23 (85%) stained positively for EGFR with a staining score of 0.05 to 1.8. Three palatal tumors attained the highest scores (one tumor, 1.2, and the remaining two, 1.8). CONCLUSIONS: Most salivary gland ACC stained positively for EGFR, and in some the staining was quite intense. On the basis of the already proven antitumoral effect of agents acting as EGFR inhibitors, it is suggested that patients with ACC might benefit from these agents, especially when surgery has failed or in those with recurrent or metastatic disease.     

Immunohistochemical study on epidermal growth factor (EGF) receptor during carcinogenesis in the rat liver.

The expression and localization of epidermal growth factor (EGF) receptor were investigated immunohistochemically using anti-EGF receptor antibody in the normal rat liver and 3'-methyl-4-dimethylaminoazobenzen (3'-Me-DAB) induced tumors in rats. In 8 weeks after 3'-Me-DAB treatment, multiple nodules of cholangiocarcinoma were found in the rat liver, and atypical nodules of hepatocytes were also found 14 weeks later. Immunoreactive products against EGF receptor were only slightly positive in the normal liver, the nodule of cholangiocarcinoma, and atypical nodule of hepatocytes. It was noted that EGF receptor immunoreactivity was more intense in non-cancerous tissue adjacent to tumorous nodules than in the cancerous tissue. The present finding suggests that the expression of EGF receptor may be associated with regenerating as well as carcinogenetic processes in the rat liver.     

前列腺癌细胞中表皮生长因子受体的表达和激活

目的：探讨前列腺癌（ＰＣａ）细胞中表皮生长因子受体（ＥＧＦＲ）表达和激活及其与雄激之间的相互作用。方法：采用免疫沉淀和Ｗｅｓｔｅｒｎ ｂｌｏｔ方法,检测雄激素依赖人ＰＣａ细胞株ＬＮＣａＰ及雄激素非依赖人ＰＣａ细胞株ＤＵ－１４５、ＰＣ－３中ＥＧＦＲ的表达和磷酸化激活,测定雄激素对ＬＮＣａＰ ＥＧＦＲ表达和激活的影响。结果：在ＤＵ－１４５和ＰＣ－３中,ＥＧＦＲ表达和激活的基础与表皮生长因子（ＥＧＦ）处     

非小细胞肺癌中上皮生长因子受体基因19位点和第21位点突变的发生与临床因素的关系

目的 探讨非小细胞肺癌中上皮生长因子受体(EGFR)19及21位点的基因突变的发生率和突变类型及其与临床因素的相关性.方法 收集115例患者的临床资料;收集该115例患者的肺癌组织(鳞癌61例,腺癌54例),对组织DNA中EGFR外显子19和21基因突变检测;分析与临床因素的相关性.结果 19和21位点突变腺癌患者突变率(45.8%)高于鳞癌患者(22.4%),非吸烟者、支气管肺泡癌的患者高.21位点突变中女性高于男性(17.3%＞9.5%,P＜0.05).结论 EGFR突变发生率约占肺癌总数的1/3,其中19位点及21位点突变分别为19.1%和13.0%,EGFR突变于非吸烟、肺腺癌和细支气管肺泡癌中突变多见,与性别无明显相关性,21位点突变于女性多见.     

Control of the epidermal growth factor receptor and its ligands during renal injury

AIM: We studied control of the epidermal growth factor (EGF) receptor and its ligands during kidney injury, since they may be importantly involved in repair. METHODS: The folic acid model of renal injury was used in these studies. Messenger RNA (mRNA) was evaluated by solution hybridization. Immunohistochemistry of transforming growth factor alpha (TGF-alpha) was also performed. RESULTS: Twenty-four hours after folic acid induced acute renal injury, creatinine increased from 0.3 +/- 0.03 mg/dl in controls to 2.0 +/- 0.8 mg/dl in folic acid injured kidneys (n = 4, p < 0.03). mRNA for the EGF receptor was increased nearly sevenfold by 24 h, and mRNA for the receptor was increased as early as 1 h following folic acid treatment. EGF receptor ligand caused a profound downregulation of the receptors in proximal tubule basolateral membranes, but receptors returned rapidly to the membrane surface in injured kidneys. The mRNA levels for heparin-binding EGF and TGF-alpha, two EGF receptor ligands, increased within 1 h of injury. TGF-alpha mRNA increased from 1.0 +/- 0.09 (relative densitometry units) in control animals to 2.9 +/- 0.13 in folic acid treated rats at 24 h (n = 4, p < 0.01), and immunohistochemical staining for TGF-alpha increased in injured kidneys at distal nephron sites. CONCLUSIONS: These studies indicate that upregulation of the EGF receptor is related to an increase in mRNA. The rapid return of receptors to the membrane surface following ligand stimulation may be useful in maintaining a mitogenic stimulus. Multiple EGF-like ligands may be important in activating the upregulated EGF receptor during repair from renal injury.     

Transactivation of epidermal growth factor receptor after heparin-binding epidermal growth factor-like growth factor shedding in the migration of prostate cancer cells promoted by bombesin

BACKGROUND: A pathway consisting of bombesin, G-protein coupling receptors (GPCRs), metalloproteases, pro-heparin-binding epidermal growth factor (proHB-EGF), and epidermal growth factor receptor (EGFR) has been reported in prostate cancer cells. The occurrence of HB-EGF shedding from proHB-EGF in this pathway, however, has not been proven directly. In addition, it is still unclear how much this pathway contributes to the migration of prostate cancer cells. In this study, we tried to directly elucidate HB-EGF shedding in this pathway and to determine its contribution to the migration of prostate cancer cells. METHODS: RT-PCR and indirect immunofluorescence staining for HB-EGF and its receptors, such as EGFR and HER4/erbB4, were performed on PC-3 cells. The influences of bombesin, anti-EGFR neutralizing monoclonal antibody, HB-EGF, and HB-EGF shedding inhibitor on the migration of PC-3 cells were studied by means of in vitro wound assays. The amount of HB-EGF shed from PC-3 cells with alkaline phosphatase-tagged HB-EGF in the presence of bombesin was determined by measuring AP activity. Immunoprecipitations and phosphotyrosine Western blotting were performed to detect EGFR transactivated by bombesin. RESULTS: PC-3 expressed HB-EGF and EGFR, but not HER4/erbB4. PC-3 migrated in the presence of bombesin, but its migration was partly inhibited by the neutralizing antibody against EGFR. PC-3 also migrated in the presence of HB-EGF, but HB-EGF shedding inhibitor partly inhibited this phenomenon. HB-EGF was shed from PC-3 cells in the presence of bombesin, and this shedding was inhibited by HB-EGF shedding inhibitor. In addition, the EGFR on PC-3 was activated in the presence of bombesin and inactivated in the presence of HB-EGF shedding inhibitor. CONCLUSIONS: These results indicated that HB-EGF shedding and the following transactivation of EGFR occurs in this pathway and that this pathway partly contributes to the migration of prostate cancer cells.     

The critical role of the epidermal growth factor receptor in endochondral ossification

Loss of epidermal growth factor receptor (EGFR) activity in mice alters growth plate development, impairs endochondral ossification, and retards growth. However, the detailed mechanism by which EGFR regulates endochondral bone formation is unknown. Here, we show that administration of an EGFR-specific small-molecule inhibitor, gefitinib, into 1-month-old rats for 7 days produced profound defects in long bone growth plate cartilage characterized by epiphyseal growth plate thickening and massive accumulation of hypertrophic chondrocytes. Immunostaining demonstrated that growth plate chondrocytes express EGFR, but endothelial cells and osteoclasts show little to no expression. Gefitinib did not alter chondrocyte proliferation or differentiation and vascular invasion into the hypertrophic cartilage. However, osteoclast recruitment and differentiation at the chondro-osseous junction were attenuated owing to decreased RANKL expression in the growth plate. Moreover, gefitinib treatment inhibited the expression of matrix metalloproteinases (MMP-9, -13, and -14), increased the amount of collagen fibrils, and decreased degraded extracellular matrix products in the growth plate. In vitro, the EGFR ligand transforming growth factor α (TGF-α) strongly stimulated RANKL and MMPs expression and suppressed osteoprotegerin (OPG) expression in primary chondrocytes. In addition, a mouse model of cartilage-specific EGFR inactivation exhibited a similar phenotype of hypertrophic cartilage enlargement. Together our data demonstrate that EGFR signaling supports osteoclastogenesis at the chondro-osseous junction and promotes chondrogenic expression of MMPs in the growth plate. Therefore, we conclude that EGFR signaling plays an essential role in the remodeling of growth plate cartilage extracellular matrix into bone during endochondral ossification.     

表皮生长因子受体在膀胱癌非肿瘤黏膜中的表达及意义

目的探讨膀胱癌非肿瘤黏膜表皮生长因子受体(EGFR)表达与膀胱癌预后的关系.方法1998-2004年原发性膀胱癌患者非肿瘤黏膜组织标本64例,标本均为距癌组织至少3 cm以上病理证实为正常膀胱黏膜;另6例正常膀胱黏膜均取自良性前列腺增生手术患者.采用S-P免疫组化染色,对检测结果进行统计学分析处理(χ2检验).结果随膀胱肿瘤病理分级增加,EGFR表达阳性表达率增高,Ⅲ级明显高于Ⅰ级和Ⅱ级(χ2=6.22,P＜0.05);EGFR表达随肿瘤临床分期的增加阳性表达率也增高,但差异无统计学意义(χ2=2.08,P＞0.05);复发性肿瘤EGFR阳性表达率明显高于未复发者,差异有统计学意义(χ2=4.95,P＜0.05).结论膀胱癌患者非肿瘤黏膜EG-FR表达较正常黏膜明显增强.EGFR在膀胱黏膜的异常表达可能早于癌细胞的出现,EGFR在膀胱肿瘤的发生、发展起着重要作用,可作为早期判断预后因子.     

K-Ras突变对表皮生长因子受体抑制剂敏感细胞株的影响

目的 观察K-Ras突变对于携带表皮生长因子受体(EGFR)突变细胞的EGFR抑制剂敏感性的影响.方法 构建K-Ras突变真核表达质粒,采用脂质体转染技术转染肺癌细胞HCC827(EGFR突变,K-Ras野生)和H292(EGFR、K-Ras均野生),噻唑蓝(MTT)比色法测定转染K-Pas突变质粒和空白质粒后各细胞对EGFR抑制剂的半数抑制浓度(IC_(50)).结果 真核表达质粒构建成功.细胞HCC827未转染K-ras突变质粒对吉非替尼(Iressa)的IC_(50)为0.007,转染后对Iressa的IC_(50)为12.3,差异有统计学意义(P<0.01).细胞H292未转染K-ras突变质粒对Iressa的IC_(50)为0.04,转染后对Iressa的IC_(50)为12.0,差异有统计学意义(P<0.01).细胞A549(K-Ras突变)对Iressa的IC_(50)为12.8,与转染K-ras突变质粒的细胞HCC827及H292比较,差异无统计学意义(P>0.05).结论 野生型或突变型EGFR出现K-Ras突变均可引起吉非替尼耐药,其程度与K-Ras突变的细胞株相当.     

胃癌组织表皮生长因子受体和下游信号的表达及临床意义

目的 检测胃癌组织中表皮生长因子受体(EGFR)、蛋白激酶B(AKT)、细胞外信号调节激酶(ERK)的表达,探讨3种蛋白在胃癌发生发展及侵袭、转移中的作用.方法 采用免疫组织化学方法(SP法)检测84例胃癌组织及15例正常胃组织中EGFR、AKT、ERK的表达,分析EG-FR、AKT、ERK的表达与胃癌患者临床病理特征及预后的关系.结果 (1)在84例胃癌组织中EG-FR、AKT、ERK过表达率分别为29.7%、31.0%和33.3%,正常胃组织的过表达均为0,差异有统计学意义(P<0.05);(2)Ⅲ+Ⅳ期患者AKT过表达率为39.7%,明显高于Ⅰ+Ⅱ期患者(11.5%)(P<0.01);(3)T3+T4组AKT的过表达率为37.9%,明显高于T1+12组(5.6%)(P<0.01);淋巴结阳性者AKT的阳性率为40.4%;明显高于阴性者(18.9%)(P<0.05).(4)EGFR、AKT、ERK过表达者总生存及无进展生存期明显低于低表达者.结论 检测胃癌组织中的EGFR、AKT、ERK表达,有助于胃癌的诊断及对胃癌的恶性程度的判断和预后的评估.     

GnRHR与EGFR在原发性肝癌组织中表达及意义

目的研究促性激素释放激素受体（GnRHR）和表皮生长因子受体（EGFR）在原发性肝癌组织中的表达及临床意义。方法应用免疫组织化SP及原位定量法,对30例原发性肝癌组织中GnRHR、EGFR的表达进行检测及半定量分析。结果在原发性肝癌各种不同分化的癌组织均显示不同程度的GnRHR和EGFR阳性免疫反应,原位定量显示GnRHR表达阳性27例（90.00%）,EGFR表达阳性15例（50.00%）,原位定量显示GnRHR的表达高分化13例、中分化9例、低分化5例,随着组织分化程度增高而增高,EGFR的表达高分化3例、中分化5例、低分化7例,随着组织分化程度增高而降低（P〈0.05）,并且GnRHR较EGFR阳性免疫反应强。结论 GnRHR与EGFR的表达量可能与原发性肝癌的发生及发展有关。     

siRNA沉默肾癌细胞EGFR基因对放疗敏感性影响的研究

目的：探讨siRNA沉默AcHN肾癌细胞的表皮生长因子受体（epidermal growth factor receptor,EGFR）对放疗敏感性的影响。方法：免疫组化及细胞免疫荧光技术证明组织水平及细胞水平EGFR的表达,化学合成针对EGFR的小干扰RNA,通过脂质体转染法转染进ACHN肾癌细胞中,利用Westernblot技术检测细胞中EGFR蛋白的表达,然后分别用0、2、4、6、8Gy剂量的x线对5组肾癌细胞（每组包含空白对照组、非异性RNA干扰组、特异性RNA干扰组）进行照射,光学显微镜下观察细胞凋亡情况,采用台盼蓝据染法检测细胞凋亡率。结果：靶向EGFR序列的特异性干扰RNA可明显抑制EGFR蛋白的表达,光学显微镜下观察到RNAi联合放疗组细胞死亡情况明显,台盼蓝拒染法检测特异性干扰EGFR基因组可显著提高AcHN肾癌细胞对放疗的敏感性（P〈O．05）。结论：体外研究表明,siRNA沉默ACHN肾癌细胞的EGFR可明显提高肾癌细胞对放疗的敏感性,为肾癌放射治疗提供了新思路。     

MUC5AC,EGFR在结石胆囊中的表达及意义

目的研究胆囊结石胆囊组织中粘蛋白5AC(mucin 5AC,MUC5AC)和表皮生长因子受体(epidermalgrowth factor receptor,EGFR)的表达,探讨胆囊结石的形成。方法采用免疫组化法和图像分析检测54例胆囊结石胆囊组织、24例对照胆囊组织(来自于壶腹部癌或胰头癌手术切除胆囊)MUC5AC,EGFR的表达情况。结果 MUC5AC和EGFR在胆囊结石胆囊组织中的表达较对照胆囊组织显著增高(P0.05)。EGFR阳性表达区域和MUC5AC阳性表达区域呈正相关(rs=0.978,P0.001)。结论 MUC5AC,EGFR可能与胆囊结石形成有关。     

富含亮氨酸重复序列免疫球蛋白2-胶质瘤细胞表皮生长因子受体信号网络新的调控靶点

目的 确定富含亮氨酸重复序列免疫球蛋白2(LRIG2)是胶质瘤细胞系GL15中EGFR信号通路新的调控靶点.方法 培养胶质瘤细胞系GL15细胞,经表皮生长因子(EGF)100μg/L,AG1478 10 μmol/L,放线菌酮(CHX)10 mg/L体外干预后,逆转录-聚合酶链反应(RT-PCR)和Western blot测定LRIG2 mRNA和蛋白表达的时间效应变化.结果 随着EGF作用时间的延长,LRIG2 mRNA先上升后恢复至正常水平.EGF作用后,表皮生长因子受体(EGFR),磷酸化EG-FR(pEGFR)均先上升后下降,LRIG2蛋白先下降,在30min时上升,120min下降至原水平的50%.CHX作用1.5 h后,再加入EGF刺激,可见LRIG2蛋白60 min降至原水平的50%.而AG1478作用1 h后,再用EGF刺激,EGFR未受明显影响,pEGFR被明显抑制,LRIG2蛋白水平变化不明显.结论 EGFR活化后可导致LRIG2 mRNA表达上升,LRIG2蛋白合成增加.