Clinical usefulness of tumour markers

Tumour markers are substances related to the presence or progress of a tumour. An ideal tumour marker is (1) detectable only when malignancy is present, (2) specific for the type and site of malignancy, (3) correlates with the amount of malignant tissue present and (4) responds rapidly to a change in tumour size. At present, no tumour marker fulfills all of the above criteria. The first part of the review discusses the clinical usefulness of the commonly requested serum tumour markers, namely, prostate-specific antigen (PSA), CA 19-9, carcinoembryonic antigen (CEA), CA 125, CA 15-3, human chorionic gonadotrophin (hCG) and alpha-foetoprotein (AFP). It is hoped that this review article will decrease the abuse and misuse of these commonly requested serum tumour markers. The second part of the review discusses the clinical usefulness of catecholamines and their metabolites, calcitonin, thyroglobulin, parathyroid hormone, prolactin, adrenocorticotrophic hormone, oestrogen and progesterone receptors, p53, HER-2/c-erbB2, BRCA1 and BRCA2.     

Clinical importance of serum anti-p53 antibodies as tumor markers

Anti-p53 antibodies are autoantibodies induced by mutation of p53 cancer-suppressor gene, and are considered to be indirect markers for p53 gene mutations and abnormally high p53 gene levels. We evaluated the usefulness of the measurement of anti-p53 antibodies by enzymed-linked immunosorbent assay using serum samples from patients with various disorders and normal subjects. The anti-p53 antibody concentration was high in patients with lung, esophageal, gastric, hepatocellular, colonic, rectal or ovarian cancer and significantly differed between the group with neoplasms and those with non-neoplastic disorders. Particularly high concentrations were observed in patients with malignant tumors. The mean agreement rate between anti-p53 antibodies and conventional tumor markers was only 47.8% despite slight differences among disorders. The positive rate increased to 63.0% by their combination assay. In addition, anti-p53 antibodies were independent markers, not complimentary to conventional markers. The mean agreement rate between anti-p53 antibodies and tissue p53 was 70.0%. Though the anti-p53 antibody-positive rate was lower than the tissue p53-positive rate, anti-p53 antibodies may be useful new tumor markers because specimens from the affected tissue are not necessary.     

Lymphangiogenic growth factors as markers of tumor metastasis

Understanding the complex process of tumor metastasis is a problem which has challenged both clinician and scientist for well over 100 years. Defining molecular markers which reflect the metastatic potential of a tumor has also proved elusive. Recently, members of the vascular endothelial growth factor (VEGF) family of glycoproteins have been demonstrated to be potent mediators of both blood vessel and lymphatic vessel formation in the context of tumor biology. Experimental studies in animal models combined with extensive clinicopathological data provide a compelling case indicating that members of the VEGF family play a key role in the formation of metastases in a broad range of solid tumors. The question of whether VEGF signaling pathways can now serve as therapeutic targets alone, or in combination with other forms of anti-cancer agents, needs to be addressed.     

Analysis of urinary mononuclear cells as markers of renal injury

The presence of macrophages(M phi) in the urine of patients with glomerulonephritis(GN) reflects the pathological events in the kidney, and we have discovered the following correlations between the M phi phenotype and the pattern of renal injury. 1) Urinary macrophage(M phi) counts increase in patients with proliferative GN, especially in the presence of active glomerular lesions(glomerular tuft necrosis, crescent, and endocapillary proliferation). In patients with hematuria, a combination of urinary M phi and T-lymphocyte counts can be used to differentiate proliferative GN from non-proliferative renal disease(hereditary nephropathy and idiopathic renal hematuria). 2) The urinary M phi of patients with active proliferative GN express Fc gamma RIII(CD16) regardless of the type of GN. 3) There are two types of urinary CD68+ cells, CD68+25F9- cells(infiltrating M phi) and CD68+25F9+ cells(mature M phi). The CD68+25F9- cell counts in the urine correlate well with the activity of proliferative GN, and the CD68+25F9+ cell counts in the urine correlate with the magnitude of non-selective proteinuria and with the subsequent decline of renal function. The CD68+25F9+ cell count increase in the urine of patients with focal segmental glomerular sclerosis, but their numbers are negligible in minimal change nephrotic syndrome. These findings indicate that analysis of the urinary M phi phenotype is a useful strategy for evaluating renal injury as a "risk-free renal biopsy".     

肿瘤标记物联合检测对肿瘤高危人群筛查的临床意义

目的 研究应用肿瘤标记物联合检测对恶性肿瘤高危人群筛查的临床价值.方法 选择血清肿瘤标记物组合对3 766例恶性肿瘤高危人员进行检测,对肿瘤标记(CA125、CEA、Cyfra21-1、CA19-9、AFP、NSE、CA72-4、SCC、CA153)升高人员进一步行颈胸部CT、腹盆部CT或内窥镜检查,女性进行妇科检查,未明确诊断者进行长期随访(大于6个月).结果 经过随访,共发现肿瘤患者69例,其中发现肿瘤标志物升高6个月内诊断为恶性肿瘤20例(62.5％),6个月至1年诊断5例(15.6％),1年至2年诊断4例(12.5％),2年至3年诊断2例(9.4％).COX分析显示肿瘤标志物升高的程度(RR=2.308,95％ CI:2.517 ～2.475,P＜0.001)、肿瘤标志物逐渐升高(RR=7.727,95％ CI:3.008 ～ 19.836,P＜0.05)和有相关临床症状(RR=7.879,95％ CI:2.357 ～ 26.384,P＜0.05)是筛查肿瘤标志物升高人员患肿瘤的危险因素.结论 血清肿瘤标记物联合检测对肿瘤筛查人群应答率高,危险因素调查是肿瘤筛查的有效手段.     

Urine diacetylspermine as a novel tumor marker

A urine tumor marker, diacetylspermine, was examined in patients with recurrent pancreato-biliary carcinoma, liver tumor, lung carcinoma and gynecologic malignancies. The urine marker increased together with recurrence, suggesting a recurrence monitoring marker at the outpatient ward. Regarding hepatocellular carcinoma, the sensitivity of the urine marker was as high as conventional markers such as AFP and PIVKA II. Synchronous examination of serum and urine markers showed a higher sensitivity than the single serum or urine marker for hepatocellular carcinoma. The sensitivity for non-advanced hepatocellular carcinoma was 50%, while that for advanced hepatocellular carcinoma was 83%. The urine marker may be useful to detect non-advanced hepatocellular carcinoma. The sensitivity for lung cancer was 83% and that for Stage I or II was 82%. Concerning uterine cervical tumor, the value of the urine marker increased with the grade of dysplasia. The sensitivity for ovarian carcinoma was 100%, while that for benign ovarian tumor was 0%. These findings suggest that urine diacetylspermine is a useful tumor marker in hepatocellular carcinoma, lung cancer and gynecologic malignancy as well as pancreatobiliary carcinoma.     

Clinical usefulness of cervicogram as a primary screening test for cervical neoplasia

The purpose of this study is to evaluate the clinical usefulness of the cervicogram as a primary screening test for cervical neoplasia. A total of 294 women who had undergone a cervicogram and a Pap test between January and July 2003, were selected. The diagnostic accuracy of the Pap test, cervicogram, and the Pap test combined with a cervicogram were compared with the histopathologic diagnosis. Among 294 women, the Pap test was negative in 130 cases and positive in 164 cases. Among patients with positive Pap test, cervicogram were negative in 101 cases (61.6%) and positive in 63 cases (38.4%). The diagnostic accuracy between cervicogram with positive Pap test and histology was as follows; sensitivity 44.9%, specificity 78.3%, positive predictive value 84.1%, negative predictive value 32.7%, false positive rate 15.9%, and false negative rate 67.3%. Although the adjunctive use of cervicogram with the Pap test in the initial screening of cervical neoplasia showed a higher specificity and higher positive predictive value compared to the Pap test alone, consideration in terms of lower sensitivity, lower positive predictive value, higher false positive rate and cost-effectiveness should be given in lieu of clinically applying cervicogram with the pap test as an initial screening test.     

Biochemical markers and physiological tests of atherosclerosis--changes and usefulness of markers in anti-atherosclerotic therapy

As a result of the aging of the Japanese population and westernization of the diet, atherosclerotic diseases such as ischemic heart diseases and cerebrovascular disorders are now the leading causes (2nd and 3rd, respectively) of death in Japan. Furthermore, advances in medical technology have made Japanese some of the longest-lived citizens in the world, and increasing health care costs have become an object of public concern. Therefore, the prevention and treatment of atherosclerosis that causes these ischemic organ dysfunctions is indispensable. In Japan, diagnostic criteria were established for metabolic syndrome as one of the health care cost-containment policies, and the importance of controlling the blood lipid and glucose levels and blood pressure was defined in 2005. Furthermore, persons enrolled in the health insurance system aged between 40 and 74 years have been obliged to receive a specified health check for the prevention of metabolic syndrome since April 2008. The health check includes: a medical interview; anthropometry (height, body weight, BMI, and abdominal circumference); physical examination; blood pressure; and 8 laboratory tests (neutral fat, HDL-cholesterol, LDL-cholesterol, AST, ALT, gamma-GT, fasting blood glucose [or HbAlc], and urinary protein and glucose). In fact, the specified health check is becoming one of the enlightening activities concerning the maintenance and promotion of health for staff of Kitasato University Hospital. Recently, on the other hand, the mechanism in which inflammatory reactions are involved in atherosclerosis has been elucidated, and atherosclerosis is thought to be a chronic inflammatory disease. Furthermore, the advances in laboratory test methods have made the measurement of various sensitive markers for inflammation possible. In this presentation, we would like to explain the efficient application of biochemical markers and rapid physiological tests for the pathological diagnosis of atherosclerosis. We would also like to explain changes in anti-thrombotic therapy aiming at the primary prevention of atherosclerosis and its usefulness.     

Analysis of novel tumor markers in pancreatic and biliary carcinomas using tissue microarrays

Using global gene expression analyses, multiple novel tumor markers overexpressed in infiltrating ductal adenocarcinomas of the pancreas have recently been identified. However, the expression of these markers in morphologically similar adenocarcinomas of the biliary tree has not been investigated. The purpose of the present study was 3-fold. First, we used 8 markers that have been shown to be overexpressed in whole tissue sections of pancreatic adenocarcinomas to validate tissue microarrays (TMAs) created from a series of pancreatic adenocarcinomas (n=68). The labeling patterns of 6 epithelial markers (fascin, mucin 4, 14-3-3sigma, prostate stem cell antigen, topoisomerase IIalpha, and cdc2/p34) were concordant with previously published studies on whole tissue sections, yet required far fewer slides and reagents. Mesothelin, an epithelial marker, and heat shock protein 47, a marker of peritumoral desmoplasia, showed lower levels of expression in the TMAs when compared with whole tissue sections. Second, we examined the previously unknown expression of the same 8 novel tumor proteins in cancers of the biliary tree by using TMAs created from a series of intrahepatic cholangiocarcinomas, gallbladder adenocarcinomas, and adenocarcinomas of the distal common bile duct (n=38). Each of the 8 markers was overexpressed in the biliary cancers, ranging from 14% demonstrating at least focal labeling with prostate stem cell antigen to 100% labeling with cdc2/p34. Most of the markers showed lower frequencies of expression in the biliary tract carcinomas in comparison to the pancreatic adenocarcinomas. In addition, expression patterns varied with location in the biliary system (intrahepatic versus gallbladder versus distal common bile duct). These differences were statistically significant (P<0.05) for mesothelin, mucin 4, and heat shock protein 47. Finally, the expression of selected markers in neoplastic progression of gallbladder cancer was examined. Two markers, fascin and mesothelin, showed up-regulation of expression with transition from carcinoma in situ to invasive adenocarcinoma, implicating a role for these markers in neoplastic progression. The results of this study indicate that TMA technology provides valid and cost-effective means to screen large numbers of novel tumor markers, even in tumors such as pancreatic and biliary adenocarcinomas that characteristically have abundant desmoplastic stroma. In addition, novel tumor markers of pancreatic adenocarcinomas show similar, yet not identical, expression patterns in biliary carcinomas. Therefore, these markers are potentially useful in developing diagnostic tests and treatment paradigms for tumors involving the biliary system.     

Endothelin receptors as novel targets in tumor therapy

The endotelin (ET) axis, that includes ET-1, ET-2, ET-3, and the ET receptors, ET_A and ET_B, plays an important physiological role, as modulator of vasomotor tone, tissue differentiation and development, cell proliferation, and hormone production. Recently, investigations into the role of the ET axis in mitogenesis, apoptosis inhibition, invasiveness, angiogenesis and bone remodeling have provided evidence of the importance of the ET-1 axis in cancer. Data suggest that ET-1 participates in the growth and progression of a variety of tumors such as prostatic, ovarian, renal, pulmonary, colorectal, cervical, breast carcinoma, Kaposi's sarcoma, brain tumors, melanoma, and bone metastases. ET-1 receptor antagonists beside providing ideal tools for dissecting the ET axis at molecular level have demonstrated their potential in developing novel therapeutic opportunity. The major relevance of ET_A receptor in tumor development has led to an extensive search of highly selective antagonists. Atrasentan, one of such antagonists, is orally bioavailable, has suitable pharmacokinetic and toxicity profiles for clinical use. Preliminary data from clinical trials investigating atrasentan in patients with prostate cancer are encouraging. This large body of evidence demonstrates the antitumor activity of endothelin receptor antagonists and provides a rationale for the clinical evaluation of these molecules alone and in combination with cytotoxic drugs or molecular inhibitors leading to a new generation of anticancer therapies targeting endothelin receptors.     

Tumor markers of urinary tract carcinoma

The tumor markers for malignant tumors arisen from urinary system including prostate cancer were reviewed. As for renal cell carcinoma there was no good marker used in routine test level at present. In the diagnosis of urothelial (transitional cell) carcinoma, mainly bladder cancer, 3 methods (urinary BTA, NMP22 and BFP) are used now in Japan. They all seem to be not fully sufficient in respect of the specificity. In foreign countries, new tests such as urinary telomerase and BLCA-4 are used and have been evaluated. On the diagnosis of prostate cancer, serum total PSA is well established and used. Various PSA relation markers have been advocated for the differentiation between benign prostate hypertrophy and carcinoma in so called "gray zone" level of total PSA. In methods based on the molecular forms of PSA, the ratio of free PSA to total PSA (f/T) is widely use, and proPSA is a test that is expected. Other approaches such as volume of index PSA, age specific PSA reference range and PSA velocity are also in practical application. Human glandular kallikrein 2, which belong to the human kallikrein family as well as PSA, is expected as a tumor specific marker.     

Clinical usefulness of wave intensity analysis

Wave intensity (WI) is a hemodynamic index, which can evaluate the working condition of the heart interacting with the arterial system. It can be defined at any site in the circulatory system and provides a great deal of information. However, we need simultaneous measurements of blood pressure and velocity to obtain wave intensity, which has limited the clinical application of wave intensity, in spite of its potential. To expand the application of wave intensity in the clinical setting, we developed a real-time non-invasive measurement system for wave intensity based on a combined color Doppler and echo-tracking system. We measured carotid arterial WI in normal subjects and patients with various cardiovascular diseases. In the coronary artery disease group, the magnitude of the first peak of carotid arterial WI (W ₁) increased with LV max. dP/dt (r = 0.74, P < 0.001), and the amplitude of the second peak (W ₂) decreased with an increase in the time constant of LV pressure decay (r = −0.77, P < 0.001). In the dilated cardiomyopathy group, the values of W ₁ were much lower than those in the normal group (P < 0.0001). In the hypertrophic cardiomyopathy group, the values of W ₂ were much smaller than those in the normal group (P < 0.0001). In mitral regurgitation before surgery, W ₂ decreased or disappeared, but after surgery W ₂ appeared clearly. In the hypertension group, the magnitude of reflection from the head was considerably greater than that in the normal group (P < 0.0001). We also evaluated hemodynamic effects of sublingual nitroglycerin in normal subjects. Nitroglycerin increased W ₁ significantly (P < 0.001). WI can be obtained non-invasively using an echo-Doppler system in the clinical setting. This method will increase the clinical usefulness of wave intensity.     

Biochemical markers of bone turnover : clinical usefulness in osteoporosis

The recent development of specific and sensitive biochemical markers reflecting the overall rate of bone formation and bone resorption, has markedly improved the non invasive assessment of bone turnover in various metabolic bone diseases, especially for osteoporosis. The immunoassay of human osteocalcin recognizing the intact molecule and its major proteolytic fragment, assays for bone alkaline phosphatase and the intact form of the N-terminal extension propeptide of type I collagen are currently the most sensitive markers to assess bone formation. The best indices of bone resorption are the new immunoassays for the pyridinoline crosslinks and type I collagen related peptides in urine, but also very recently available in serum. Using these new markers, several studies have shown that bone turnover increases markedly after the menopause and remains elevated in late postmenopausal and elderly women. An increased bone turnover rate is related to a fast rate of bone loss in postmenopausal women and to a decreased bone mass in elderly women. Recent data suggest that some of the new immunoassays for pyridinoline crosslinks and related peptides could predict the subsequent risk of hip fracture in elderly women. Thus, bone markers might be used in combination with bone mass measurement to improve the prognostic assessment of postmenopausal women, i.e. their risk of developing osteoporosis and ultimately fractures. Treatment of postmenopausal women with antiresorptive drugs such as estrogens, bisphosphonates and calcitonin is followed by rapid decrease of the levels of bone markers that is correlated with the long term increase of bone mass as assessed by dual-energy X ray absorptiometry measurement. Thus, bone markers should be useful in monitoring treatment efficacy in patients with osteoporosis. Appropriate combination of the most efficient markers of bone formation and resorption will probably provide a powerful tool in the clinical investigation of osteoporosis.     

A novel urinary sialoglycoprotein as the inhibitor of interleukin-1.

An inhibitor of interleukin-1 (IL-1) was purified to homogeneity from febrile human urine by using a sequence of ammonium sulphate fractionation, DEAE-cellulose chromatography and gel filtration on a column of ACA 54. The inhibitor was a sialoglycoprotein and its molecular weight, examined by SDS-PAGE, was found to be 30 kD. It was acidic in nature and its isoelectric point, determined by electrofocusing on thin-layer polyacrylamide gels, was found to be 3.5-3.6. The inhibitor was sensitive to the action of sialidase from Vibrio cholerae as the enzyme treated material was electrofocused at a higher pH range (4.5-5.2). The inhibitor contained 10% sialic acid as estimated by the Thiobarbituric acid assay. It interacted with the lectins jacalin and concanavalin A, suggesting the presence of carbohydrate moieties such as galactose and mannose. The inhibitor did not block the specific binding of IL-1 to cells, nor did it directly bind IL-1. Early rejection urine from a kidney transplant patient was also found to contain the 30 kD IL-1 inhibitor protein. This was visualised by immunoblotting using specific antibody raised against the 30 kD IL-1 inhibitor isolated from the urine of febrile patients.     

Urinary nucleosides as potential tumor markers evaluated by learning vector quantization

Modified nucleosides were recently presented as potential tumor markers for breast cancer. The patterns of the levels of urinary nucleosides are different for tumor bearing individuals and for healthy individuals. Thus, a powerful pattern recognition method is needed. Although backpropagation (BP) neural networks are becoming increasingly common in medical literature for pattern recognition, it has been shown that often-superior methods exist like learning vector quantization (LVQ) and support vector machines (SVM). The aim of this feasibility study is to get an indication of the performance of urinary nucleoside levels evaluated by LVQ in contrast to the evaluation the popular BP and SVM networks. Urine samples were collected from female breast cancer patients and from healthy females. Twelve different ribonucleosides were isolated and quantified by a high performance liquid chromatography (HPLC) procedure. LVQ, SVM and BP networks were trained and the performance was evaluated by the classification of the test sets into the categories "cancer" and "healthy". All methods showed a good classification with a sensitivity ranging from 58.8 to 70.6% at a specificity of 88.4-94.2% for the test patterns. Although the classification performance of all methods is comparable, the LVQ implementations are superior in terms of more qualitative features: the results of LVQ networks are more reproducible, as the initialization is deterministic. The LVQ networks can be trained by unbalanced sizes of the different classes. LVQ networks are fast during training, need only few parameters adjusted for training and can be retrained by patterns of "local individuals". As at least some of these features play an important role in an implementation into a medical decision support system, it is recommended to use LVQ for an extended study.     

肿瘤标记物检测在肺部孤立性结节鉴别诊断中的价值

为研究外周血中肿瘤标志物癌胚抗原(CEA)、神经元特异性烯醇化酶(NSE)、细胞角质蛋白19片段(CYFRA21-1)、糖链抗原125(CA125)、鳞状上皮细胞癌抗原(SCC)检测在肺部孤立性结节鉴别诊断中的价值,选取上海市胸科医院2013年9月至2015年1月295例因肺部孤立性结节入院患者的血清标本,其中恶性结节患者221例,良性结节患者74例,采用化学发光法检测患者血清中SCC的表达水平,采用流式荧光法检测血清中CEA、NSE、CA125和CYFRA21-1的表达水平,评价现有肿瘤标志物在良、恶性肺部孤立性结节鉴别诊断中的价值。结果显示,良性和恶性肺部孤立性结节患者血清中CA125和SCC的浓度无明显差异,没有统计学意义(P0.05);而恶性结节患者血清中CEA,NSE和CYFRA21-1的检测水平显著高于肺部良性结节患者,差异有统计学意义(P0.05)。在肺部孤立性结节实验室鉴别诊断中,CEA的灵敏度为59.7%,特异度为81.1%;NSE的灵敏度为36.7%,特异度为85.1%;CYFRA21-1的灵敏度为50.2%,特异度为70.3%;三者联合检测灵敏度达60.2%,特异度达85.1%。以上结果说明,NSE、CYFRA21-1和NSE的联合检测在肺部孤立性结节鉴别诊断中具有一定的临床价值。