A carcinoembryonic antigen-secreting adenocarcinoma arising in tailgut cyst: clinical implications of carcinoembryonic antigen

Tailgut cysts (TGCs) are rare congenital cysts that occur in the retrorectal or presacral spaces. Although most tailgut cysts have been reported as benign, there have been at least 9 cases associated with malignant change. We report herein on an unusual case of a 40-year-old woman with a carcinoembryonic antigen (CEA)-producing adenocarcinoma arising within a TGC who underwent surgical resection and local radiation therapy. Despite the complete resection, metastatic adenocarcinoma developed five months after surgery. CEA-producing adenocarcinoma from a TGC is extremely rare and only two cases, including this case, have been reported in the English medical literature. Besides CEA, the serum levels of CA 19-9 became markedly elevated in this patient. Given that the serum CEA level decreased to the normal range after complete resection of tumor and that the tumor recurrence was associated with a rebound of the CEA serum level, our case shows that serial measurements of serum CEA can be used for treatment planning and for assessing the patient's treatment response for this rare disease.     

Immunoelectron microscopic localization of carcinoembryonic antigen in gastric adenocarcinoma cell lines

The distribution of carcinoembryonic antigen (CEA) in human gastric adenocarcinoma cell lines (HPE-GAC-3 cells and HPE-GAC-2 cells) was determined immunohistochemically by indirect peroxidase-labeled antibody method at the light and electron microscopic levels. In GAC-3 cells that proliferated as non-adherent single cells, CEA was located in the perinuclear spaces, the endoplasmic reticulum, Golgi apparatus, vesicles, multivesicular body (MVB) and entire plasma membrane. Membrane CEA was shown to be internalized into MVB in GAC-3 cells. In GAC-2 cells that form an acinus, CEA was predominantly present along the microvilli of the lumina) surface and in glycocalyceal bodies, the vesicles which bud from the microvilli into the lumen. These results suggest that in poorly differentiated cancer cells CEA is transported over the entire cell surface, retained on the membrane and accumulated Into the cell by way of the MVB, but in well differentiated cancer cells the newly synthesized CEA is rapidly and predominantly transported to the luminal surface and rapidly released from the membrane into the lumen by way of the glycocalyceal body.     

Metastatic potential of human colorectal carcinoma SW1222 cells transfected with cDNA encoding carcinoembryonic antigen

In order to examine a role of carcinoembryonic antigen (CEA) in metastasis, cDNA encoding CEA was introduced into a clone of human colorectal carcinoma SW1222 cells. Western blot analysis revealed that all transfectants express CEA of 180 kDa while the parent clone does not. In the transfectants, the level of CEA expression in clone 3 was higher than that of clone 1. Clone 3 formed aggregates rapidly after suspended by trypsinization while clone 1 did not. In experimental metastasis assay where tumor cells were injected intrasplenically, clone 3 exhibited a higher liver-metastatic activity than clone 1. Fab fragment of anti-CEA antibody significantly inhibited both the cell aggregation and the liver metastases caused by clone 3. These findings suggested that CEA expressed on the cell surface may play an important role in hepatic metastasis from colorectal carcinoma, possibly through its cell adhesion activity.     

Endocervical tubal metaplasia and adenocarcinoma in situ: role of immunohistochemistry for carcinoembryonic antigen and vimentin in differential diagnosis

Eighteen cases of endocervical tubal metaplasia (tubal metaplasia) and 16 cases of adenocarcinoma in situ were reacted immunohistochemically for carcinoembryonic antigen (CEA) and vimentin. Whereas CEA was more frequently positive in adenocarcinoma in situ cases (63%) than in tubal metaplasia cases (39%), vimentin was not expressed in adenocarcinoma but was positive in 78% of cases of tubal metaplasia.     

Immunohistological study of tight junction protein expression in mal de Meleda

Mal de Meleda (MdM, MIM: 248300) is a rare autosomal recessive skin disorder characterized by diffuse palmoplantar keratoderma and transgressive keratosis with onset in early infancy. The gene responsible for MdM, ARS, encodes for Secreted Lys6/Plaur domain-containing protein 1 which is essential for epidermal homeostasis. Tight junctions have been proposed to have two mutually exclusive functions: a fence function which prevents the mixing of membrane proteins between the apical and basolateral membranes; and a gate function which controls the paracellular passage of ions and solutes between cells. In this study we report immunohistochemical investigations of tight junction proteins claudin-1 and occludin in MdM Tunisian families. Nine skin biopsies from patients with MdM were analyzed. The control group was formed by skin biopsies belonging to healthy individuals. Immunohistochemical study was performed on fixed sections from biopsies of four microns with the following polyclonal antibodies: anti-claudin-1 and anti-occludin. In control skin, claudin-1 exhibited membrane expression throughout the epidermis with increasing and upward intensity, whereas occludin was detected in the cell membrane of keratinocytes of the stratum granulosum. In MdM skin, claudin-1 was expressed throughout the thickness of the spinous layers with membrane staining, and occludin had cytoplasmic staining in the granular layer. The immunohistochemical expression of TJ proteins in MdM patients harbors premature expression of occludin and decreased expression of claudin-1, highlighting further evidence for disorders in epidermal homeostasis.     

Alterations in tight junction molecules of uterine epithelial cells during early pregnancy in the rat

Distribution patterns of the tight junction associated proteins ZO-1, claudin-1 and occludin were investigated in rat uterine epithelial cells during early pregnancy. Light microscopy and immunohistochemical labelling were used to detect these proteins on days 1, 3, 6 and 7 of pregnancy. Intense staining of claudin-1 at the apical region of the lateral plasma membrane accompanied diffuse staining throughout the cytoplasm. ZO-1 was also localised in the apical region, but ZO-1 was not present in the lower two thirds of the lateral plasma membrane or in the cytoplasm. Occludin was present only on days 6 and 7 of pregnancy. Labelling was also localised in the apical region of the lateral plasma membrane where tight junctions are known to be present. Our results show that ZO-1, claudin-1 and occludin are present in the apical region of uterine epithelial cells, and appear to play a role in the very dynamic tight-junctional network of uterine epithelial cells during early pregnancy. In particular, occludin appears only during uterine receptivity for implantation.     

抗CEA嵌合小分子抗体的构建与表达

目的 构建表达抗CEA小分子抗体,以利于放射免疫诊断及导向治疗。方法 构建CEA单链抗体基因,克隆入载体pKpL-3a,在大肠杆蓖中包涵体表达,ELISA检测活性,将轻重链基因克隆入分泌型表达载体pSCMH,在大肠杆菌中表达。ELISA检测活性,钭CEA嵌合抗体的轻链和Fd基因克隆入杆状病毒表达载体pAcUW51中,在sf9细胞中分泌表达,ELISA检测活性及测定产量,竞争抑制ELISA测定其识别的抗原表位。结果 多种方法复性的包涵体及原核分泌表达的单链相同的表位。结论 该CEA抗体基因在大肠杆菌中不能表达出有功能的分子。而在昆虫细胞中表达了具自学成才性抗体分子,某些抗体基因只有在真核细胞中才能表达出有功能的抗体分子。     

Sandwich enzyme immunoassay using three monoclonal antibodies against different epitopes of carcinoembryonic antigen (CEA)

Purified monoclonal antibodies (Mab) produced by 3 hybridomas and reacting with 3 different epitopes of carcinoembryonic antigen (CEA) were used in a solid phase enzyme immunoassay. Two Mabs were physically adsorbed to polystyrene balls and the third Mab was coupled to alkaline phosphatase using the bifunctional reagent N-succinimidyl-3-(2-pyridyldithio)-propionate. During a first incubation, CEA from heat-extracted serum samples was immunoadsorbed to the antibody coated balls. After washing of the balls, bound CEA was detected by a second incubation with the enzyme coupled Mab. The sensitivity of the assay was 0.6 ng per ml of serum. A total of 196 serum samples from patients with various types of carcinoma, with liver cirrhosis, or from healthy blood donors with or without smoking habits, were tested. The results obtained with the monoclonal enzyme immunoassay (M-EIA) were compared with those obtained with perchloric acid extracts of the same serum samples tested by an inhibition radioimmunoassay using conventional goat anti-CEA antiserum. There was an excellent correlation between the two assays. In particular, the new M-EIA gave good results for the detection of tumor recurrences in the follow-up of colon carcinoma patients. However, despite the use of exclusively monoclonal antibodies the new assay detected a similar percentage of slightly elevated CEA values as the conventional assay in patients with non-malignant disease, suggesting that the CEA associated with non-malignant diseases is immunologically identical to the CEA released by colon carcinoma.     

Value of thrombomodulin immunostaining in the diagnosis of transitional cell carcinoma: a comparative study with carcinoembryonic antigen

Aims: Thrombomodulin (TM) is a surface glycoprotein involved in the regulation of intravascular coagulation that has been reported to be expressed in a variety of tumours. We investigated TM expression in transitional cell carcinoma (TCC) and compared the value of TM immunostaining with that of carcinoembryonic antigen (CEA) for differentiating TCC from other tumours with which it may be confused.  

Methods and results:

Immunostaining was performed on formalin-fixed, paraffin-embedded tissue sections using the avidin–biotin–peroxidase complex method. TM immunoreactivity was observed in 80 of 91 primary (51/58 urinary bladder, 10/12 renal pelvis, 3/3 ureter, 15/15 prostate, 1/3 ovary), and 18 of 20 metastatic TCCs expressed this marker. Only 37 of the 91 primary (23/58 urinary bladder, 4/12 renal pelvis, 1/3 ureter, 9/15 prostate, 0/3 ovary) and six of the 20 metastatic TCCs reacted for CEA. In order to evaluate the practical utility of TM immunostaining in surgical pathology, 30 adenocarcinomas of the prostate, 18 of the bladder, 12 of the colon, and 22 renal cell carcinomas were also stained for these markers. CEA reactivity was obtained in 12 of 30 adenocarcinomas of the prostate, 12 of 18 of the bladder, and 12 of 12 of the colon, but in none of the 22 renal cell carcinomas. Only three of the 18 adenocarcinomas of the bladder showed focal TM reactivity, but no staining for this marker was observed in any of the other types of tumours.  

Conclusions:

TM is a more sensitive marker than CEA for TCC and, because it has a more restricted reactivity with other tumours, TM has more practical value in separating TCCs from adenocarcinomas of the prostate, colon and bladder, and renal cell carcinomas than CEA.     

Expression of occludin in human rectal carcinoid tumours as a possible marker for glandular differentiation

AIMS: To examine whether or not the tight junction-associated transmembrane protein occludin is expressed in rosette or gland-like structures in human rectal carcinoid tumours. The tight junction is crucial for the formation and maintenance of organized tubular structures in glandular epithelia. Previous studies have reported the presence of glandular structures in carcinoid tumours, though they are not believed to arise from glandular epithelium. METHODS AND RESULTS: The expression profiles of occludin in 40 carcinoid tumours were examined immunohistochemically, using an anti-occludin monoclonal antibody. In eight (20%) samples of typical carcinoid tumours, a small number of rosette-like tubular structures outlined by occludin were detected. CONCLUSIONS: Tight junction-associated molecules, including occludin, are thought to be one of the most characteristic structural markers of polarized glandular structures. The results of the present study provide supportive evidence that carcinoid tumour cells are capable of glandular differentiation.     

Inhibition of endogenous carcinoembryonic antigen (CEA) increases the apoptotic rate of colon cancer cells and inhibits metastatic tumor growth

It has been suggested that carcinoembryonic antigen (CEA) enhances metastatic seeding of colon cancer cells due to its homo- and heterophilic binding properties. Our recent finding that endogenous CEA protects colon cancer cells against apoptosis suggests a more complex role of CEA in cancer progression. In this study we compared the in vitro effects of endogenous CEA on tumor cell aggregation and cell cycle regulation of human HT29 colon cancer cells with the corresponding in vivo effects, i.e. tumor cell seeding and formation of metastatic lesions. Stable expression of CEA targeted ribozymes (Rz) under control of a tet-off promoter system allowed regulation of CEA levels on the mRNA and protein level by 50%. Downregulation of CEA levels inhibited tumor cell aggregation by 70%. In accordance with previous studies [1], reduction of CEA levels increased in vitro the apoptotic rate and reduced colony formation by 30% to 50%. To determine the in vivo effect of CEA-dependent aggregate formation and its growth regulating role under apoptotic stress, HT29 cells with high and low CEA levels, respectively, were injected into nude mice. Immunostaining of lung microsections revealed similar numbers of tumor cells one hour after injection. 24 h later virtually all cells were removed from the lung in both groups. However, after 6 weeks all doxycycline treated mice (Rz off = CEA high) showed 14.5 ± 4.6 metastatic lung lesions/mouse while 0.2 ± 0.2 lesions/mouse appeared in the untreated group (Rz on = CEA low) ( P<0.001). Our study demonstrates a multifunctional role of CEA and indicates a prometastatic role of CEA independent of its adhesive function possibly due to its anti-apoptotic function. This revised version was published online in July 2006 with corrections to the Cover Date.     

聚类分析法在癌胚抗原数据挖掘分析中的应用研究

目的探讨聚类分析方法在癌胚抗原数据的应用,以获取有价值的诊断信息。方法回顾分析自2008年4月至2013年4月癌胚抗原数据,采用SPSS Clementine数据挖掘软件的K-means模型,对CEA相关的性别、年龄等特征用聚类分析法进行挖掘,分析癌胚抗原及关联特征的临床意义。结果K-means算法对12532行癌胚抗原数据的挖掘结果分5组,其中3组有医学意义,特别是平均年龄为88.541岁的高龄男性老年患者罹患癌症的概率,较老年女性及低龄老年男性更高,临床上应加大对该部分人群的体检力度,争取早发现早治疗。结论通过对血清癌胚抗原结果进行数据挖掘,发现聚类结果符合医学意义,可实现精准的预防及治疗。     

Expression patterns of tight junction proteins in porcine major salivary glands: a comparison study with human and murine glands

Tight junction (TJ) proteins play a dynamic role in paracellular fluid transport in salivary gland epithelia. Most TJ studies are carried out in mice and rats. However, the morphology of rodent salivary glands differs from that of human glands. This study aimed to compare the histological features and the expression pattern of TJ proteins in porcine salivary glands with those of human and mouse. The results showed that porcine parotid glands were pure serous glands. Submandibular glands (SMGs) were serous acinar cell‐predominated mixed glands, whereas sublingual glands were mucous acinar cell‐predominated. Human SMGs were mixed glands containing fewer mucous cells than porcine SMGs, whereas the acinar cells of murine SMGs are seromucous. The histological features of the duct system in the porcine and human SMGs were similar and included intercalated, striated and excretory ducts, but the murine SMG contained a specific structure, the granular convoluted tubule. TJ proteins, including claudin‐1 to claudin‐12, occludin and zonula occludin‐1 (ZO‐1), were detected in the porcine major salivary glands and human SMGs by RT‐PCR; however, claudin‐6, claudin‐9 and claudin‐11 were not detected in the murine SMG. As shown by immunofluorescence, claudin‐1, claudin‐3, claudin‐4, occludin and ZO‐1 were distributed in both acinar and ductal cells in the porcine and human SMGs, whereas claudin‐1 and claudin‐3 were mainly present in acinar cells, and claudin‐4 was mainly distributed in ductal cells in the murine SMG. In addition, 3D images showed that the TJ proteins arranged in a honeycomb‐like structure on the luminal surface of the ducts, whereas their arrangements in acini were irregular in porcine SMGs. In summary, the expression pattern of TJ proteins in salivary glands is similar between human and miniature pig, which may be a candidate animal for studies on salivary gland TJ function.     

血清CEA水平在EGFR-TKI治疗非小细胞肺癌的应用价值

目的：探讨血清癌胚抗原(carcino-embryonic antigen,CEA)在表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)治疗非小细胞肺癌的价值。方法：回顾性收集确诊非小细胞肺癌并使用EGFR-TKI治疗患者的治疗前后CEA水平、一般资料、病理亚型、分期、分子标记物和CEA对治疗决策的影响,并定期影像学评价,随访至进展生存期(progression-free survival,PFS)和总生存期(overall survival,OS)。通过Kaplan-Meier绘制生存曲线,log-rank检验比较不同CEA组间的生存差异, Cox回归分析预后因子。结果：本研究纳入2002年6月至2011年8月广东省肺癌研究所使用EGFR-TKI的非小细胞肺癌患者209例。分为3组：L组(CEA≤5,n=54),M组(520,n=103)。L组、M组和H组的中位PFS分别为19、17和13月(95% CI,14.61~19.39;P=0.018);中位OS分别为32、31和24月(95% CI,25.14~30.86;P=0.010)。治疗前低CEA的患者有显著延长的PFS和OS。CEA升高同时伴有影像学进展的45例,CEA升高无影像学进展证据的27例,两组的中位PFS分别为13月和14月(95% CI,10.51~15.49,P=0.195);中位OS分别为23月和28月(95% CI,19.81~30.19,P=0.156)。CEA升高无影像学进展的27例患者均随访至进展才改变治疗策略,中位无进展时间为24.3月(3~111月)。结论：治疗前血清CEA水平和EGFR-TKI治疗晚期非小细胞肺癌的预后呈负相关;治疗过程中CEA升高时,需结合影像学和症状来决定是否改变治疗策略。     

The immunohistological demonstration of carcinoembryonic antigen in intra-epithelial and invasive squamous carcinoma of the cervix

Using the sensitive peroxidase, anti-peroxidase immunohistological technique, carcinoembryonic antigen (CEA) was studied in formalin-fixed, paraffin-embedded cervical specimens from 225 patients. CEA was demonstrated in 33% of moderate dysplasia, 78.5% of severe dysplasia, 79% of in-situ carcinoma and 78% of invasive carcinoma. CEA was not demonstrated in normal squamous epithelium, squamous metaplasia, reserve cell hyperplasia nor in mild dysplasia.     

Monoclonal antibodies against carcinoembryonic antigen (CEA) used in a solid-phase enzyme immunoassay. First clinical results

A solid-phase enzyme immunoassay using both mouse monoclonal and goat polyclonal antibodies against carcinoembryonic antigen (CEA) was developed. The assay detects 0.6 to 1.2 ng of CEA per ml of serum and has 3 incubation steps which can be performed in 1 day. Polystyrene balls coated with polyclonal goat anti-CEA antibodies are first incubated with heat-extracted serum samples. Bound CEA is then detected by addition of mouse monoclonal antibodies, followed by goat IgG anti-mouse IgG1 coupled to alkaline phosphatase. Results with this enzyme immunoassay using monoclonal antibodiies (M-EIA) have been compared with those obtained by the conventional inhibition radioimmunoassay (RIA) using goat antiserum. Three hundred and eighty serum samples from 167 patients with malignant or non-malignant diseases and from 134 normal individuals with or without heavy smoking habits were analyzed by the 2 assays. Excellent correlation between the results of the 2 assays was obtained, but the M-EIA, using monoclonal antibodies from a single hybridoma, did not discriminate better than the conventional RIA between CEA produced by different types of carcinoma and between CEA associated with malignant or non-malignant diseases. Follow-up studies of several patients by sequential CEA determinations with the 2 assays showed that the M-EIA was as accurate as the RIA for the detection of tumor recurrences.     

Correlation of carcinoembryonic antigen in tissue sections with spread of mammary carcinoma

An immunoperoxidase technique for the histological demonstration of Carcinoembryonic Antigen (CEA) was applied to paraffin sections from 74 breast carcinomas and 43 benign breast lesions. Sixty-six per cent of the carcinomas and the only case of granular cell myoblastoma examined were CEA positive. Two examples of mammary dysplasia (7%) showed foci of CEA positive acini. All tumour tissue found in lymphatics and in metastases in lymph nodes was CEA positive, including two cases where the primary tumour was CEA negative, and all the metastases examined from CEA positive tumours. A significant relationship was found between CEA positivity of the primary tumour and the presence of lymph node metastases.     

血清癌胚抗原、糖链抗原153与异常糖链糖蛋白在乳腺癌诊断中的应用

目的:探讨肿瘤异常糖链糖蛋白(tumor abnormal protein,TAP)、癌胚抗原(carcinoembryonic antigen,CEA)与糖链抗原153(carbohydrate antigen 153,CA153)在乳腺癌早期诊断中的临床价值.方法:收集45例乳腺癌患者为研究组,49例正常体检人群为对照组,比较各肿瘤志记物的敏感度、特异度、准确度等指标.结果:在乳腺癌组患者中,TAP,CEA,CA153检测敏感度分别为80.00％,8.89％,6.67％;特异度分别为100.00％,97.96％,100.00％,准确度分别为90.42％,55.32％,55.32％.结论:肿瘤TAP检测优于肿瘤标志物CEA和CA153检测,可作为乳腺癌的早期筛查指标.     

血清甲胎蛋白联合癌胚抗原检测在肝癌诊断中的应用价值（附125例报告）

目的探讨血清甲胎蛋白联合癌胚抗原检测在原发性肝癌和转移性肝癌诊断中的应用价值。方法回顾性分析我院2003年7月～2006年3月收治的125例肝癌患者的临床资料,同时测定他们和65例健康人血清中的血清甲胎蛋白和癌胚抗原含量。结果肝癌组血清甲胎蛋白和癌胚抗原含量及阳性率明显高于正常对照组,且转移性肝癌中癌胚抗原含量明显高于原发性肝癌。结论检测血清中血清甲胎蛋白和癌胚抗原的含量,对肝癌的诊断有很高的价值,对鉴别肝癌的类型具有一定的指导意义。     

大鼠脑缺血时紧密连接相关蛋白occludin和claudin-5表达的变化

目的研究脑缺血后血脑屏障通透性和紧密连接相关蛋白occludin和claudin-5的变化。方法线栓法制备大鼠大脑中动脉缺血模型,采用伊文氏兰(EB)法检测缺血后血脑屏障通透性的变化。应用免疫组织化学的方法观察occludin和claudin-5在缺血脑组织中的分布,采用RT-PCR、Wesrern blot法检测大鼠缺血脑组织occludin和claudin-5的mRNA和蛋白的表达变化。结果脑缺血2h后血脑屏障通透性显著增加(P0.01),紧密连接相关蛋白occludin和claudin-5在脑微血管内皮细胞上呈阳性表达,occludin和claudin-5的mRNA和蛋白均比正常对照组显著降低(P0.01)。结论脑缺血时血脑屏障通透性的增加可能与紧密连接相关蛋白occludin和claudin-5的降低相关。