Serum alpha-fetoprotein predicts treatment outcome in chronic hepatitis C patients regardless of HCV genotype

We examined the association between serum alpha-fetoprotein (AFP) level and sustained virological response (SVR) in 93 chronic hepatitis C patients. The SVR rate was much higher among patients with serum AFP levels below rather than above the median value (5.7 ng/ml) (58.7% and 19.2%, respectively; P<0.0001). Serum AFP should be added to the list of factors predictive of treatment response in chronic hepatitis C.     

Alpha-fetoprotein synthesis during liver regeneration in adult mice of different strains]

Alpha-fetoprotein (AFP) concentration was estimated by radial immunodiffusion in agar technique in the sera of adult mice of 12 inbred strains and F1 hybrids (SWR X B10D2 and B10D2 X SWR) during liver regeneration after CCl4 poisoning. Statistically significant difference in the AFP concentration was found in the male and female sera in 6 of 10 mouse strains studied. The following inter-strain differences were also revealed: the AFP mean levels in the sera of C57BL/6 and B10D2 strains were found to be significantly lower than the corresponding levels in the sera of the majority of the strains tested. F1 hybrids showed to occupy an intermediate position by the AFP content between the parental strains. Small, but statistically significant, differences were revealed between the groups of male F1 hybrids from the direct reciprocal crossings. It is suggested that the control of induction of the AFP synthesis during the liver regeneration in adult mice was realized on the polygene basis.     

Immunoperoxidase labelling of alpha1-fetoprotein (AFP) in normal and regenerating livers of a low and a high AFP producing mouse strain

Summary Serum AFP concentrations in normal adult BALB/c/J and in normal adult C3H/He mice were in the order of 0.6 g/ml and 0.1 g/ml, respectively. In BALB mice, AFP was localized in the cytoplasm of differentiated mono- and binucleated hepatocytes in centrolobular and intermediate zones of normal adult liver. No cellular AFP could be detected in liver sections of normal adult C3H mice.CCl₄ intoxication was accompanied by increase of serum AFP levels. A maximum was reached on day 4. Afterwards, concentrations declined. In sera of BALB/c/J mice, AFP levels reached values 10-fold higher and more than in sera of C3H/He mice.From day one after CCl₄ intoxication, cellular AFP was detected in hepatocytes of portal and periportal areas including intermediate zones adjacent to the necrosis. The intensity of AFP staining reached a maximum between the days 3 and 4. Hepatocytes in front of the necrotic areas usually contained the strongest AFP reactions. In both mouse strains, cellular AFP pattern was comparable, but strongest immunoreactivity was observed in liver sections of BALB/c/J mice.Liver injury and subsequent regeneration occurred to the same extent in both studied strains. The much higher serum AFP levels and the stronger AFP immunolocalizations in BALB mice were thought not due to increased numbers of AFP producing and releasing cells during liver regeneration. Additional mechanisms must play a role in increased AFP synthesis per single cell. C3H/He was a low AFP-inducible and BALB/c/J was a high AFP-inducible mouse strain.Supported by the Deutsche Forschungsgemeinschaft (Ku 257/3) Bonn. Federal Republic of Germany     

Estrogen induction of progestophilins in rat estrogen-sensitive cells grown in media supplemented with sera from castrated rats and from rats bearing an alpha-fetoprotein-secreting hepatoma

The purpose of this work was to study the effect of alpha-fetoprotein (AFP) over cell multiplication and the induction of an estradiol-17 beta (E2)-dependent marker, i.e., progestophilins in E-sensitive cells C2(9)RAP derived from a W/Fu rat pituitary tumor. These cells proliferate in isogeneic hosts under the influence of E2, while they proliferate in culture regardless of the presence of E2. C2(9)RAP cells were grown in medium supplemented with 10% horse serum. Progestophilin levels were measured 48 h after adding serum (20% horse, or castrated rat, or AFP-secreting tumor-bearing rat) and estrogen to the 10% horse serum-supplemented medium in which the cells were growing. Maximal induction of progestophilins was obtained at 3 X 10(-10) M E2 in cells grown in medium containing horse or castrated rat serum. In contrast, maximal induction of progestophilins required 3 X 10(-8) M E2 in cells grown in medium supplemented with the serum of Morris hepatoma 7777-bearing rats. This serum contained AFP levels comparable to those present at birth in the rat. 11-Methoxy-17 beta ethynylestradiol (R2858), a synthetic estrogen with little affinity for AFP, was also tested for its ability to induce progestophilins. The degree of maximal induction of progestophilins expressed as percentage of the respective control, was similar for all experimental groups, both with E2 and with R2858. In addition, we compared the free E2 levels in the culture medium with the progestophilin levels and the cell proliferation rate. We found that the progestophilin levels were maximal at free E2 concentrations above 11 pg E2/ml, whereas there was no correlation between the free E2 levels and the proliferation rate. Moreover, the proliferation rate of cells in medium supplemented with horse or castrated rat serum was maximal at concentrations of free E2 below 0.4 pg/ml; whereas cell proliferation was inhibited with hepatoma serum even at concentrations of free E2 of 44 pg/ml. We conclude that the effect of hepatoma serum on the E2 induction of progestophilins seems to be mediated by the effect of AFP on the availability of free estrogen, since it is abolished by the addition of both natural and synthetic estrogens. The inhibitory effect of hepatoma serum upon cell proliferation is not reversed by estrogens and thus seems to be mediated by mechanisms other than E2 trapping by AFP.     

Alpha-fetoprotein in Abortion

The clinical value of maternal serum alpha-fetoprotein (AFP) as a guide to the outcome of threatened abortion was assessed. After the thirteenth week of gestation, abortion occurred more frequently (10/12) in women with abnormal serum AFP levels than in those (2/12) whose AFP concentrations were within the normal range. Low levels were present in women with blighted ovum and high concentrations were associated with intrauterine fetal death. In legal first and second trimester abortions, the circulating maternal AFP levels in postabortion samples were often higher than before abortion, irrespective of whether abortion was performed instrumentally or induced with prostaglandins. Maternal serum AFP levels provide a new means for prediction of the outcome of threatened abortion.     

Binding of 16α-[18F]Fluoro-17β-estradiol to alphafetoprotein in Sprague-Dawley female rats affects blood levels

To examine the relationship between blood levels of 16α-[¹⁸F]fluoro-17β-estradiol(¹⁸F-ES) and serum alphafetoprotein (AFP) concentration, we undertook a study in which serum from various aged (20–33 days old) Sprague-Dawley female rats injected with ¹⁸F-ES was analyzed for both blood activity levels and AFP. There is a strong positive correlation between serum AFP concentration and ¹⁸F-ES blood levels (r = 0.914, P < 0.001), suggesting that the binding of ¹⁸F-ES by AFP has a significant effect on blood activity levels. The AFP concentration and ultimately the AFP-¹⁸F-ES binding is dependent on the age and weight of the rat: younger, as well as low weight rats exhibited high AFP concentrations and consequently increased ¹⁸F-ES blood activity. The rats most suitable for comparative studying of labeled estrogens are 25–28 days of age and weigh a minimum of 50–55 g. Thus, the use of the immature rat model to compare labeled estrogens requires a careful consideration of possible interference from blood binding proteins (i.e. AFP), as well as potential receptor binding competition from endogenous estrogens produced during the estrous cycle. Comparable consideration of blood binding protens (sex steroid binding protein, SBP) and endogenous estrogens must be made in human studies, as well.     

Quantitative proteomic signature of liver cancer cells: tissue transglutaminase 2 could be a novel protein candidate of human hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common diseases worldwide, with extremely poor prognosis due to failure in diagnosing it early. Alpha-fetoprotein (AFP) is the only available biomarker for HCC diagnosis; however, its use in the early detection of HCC is limited, especially because about one-third of patients afflicted with HCC have normal levels of serum AFP. Thus, identifying additional biomarkers that may be used in combination with AFP to improve early detection of HCC is greatly needed. A quantitative proteomic analysis approach using stable isotope labeling with amino acids in cell culture (SILAC) combined with LTQ-FT-MS/MS identification was used to explore differentially expressed protein profiles between normal (HL-7702) and cancer (HepG2 and SK-HEP-1) cells. A total of 116 proteins were recognized as potential markers that could distinguish between HCC and normal liver cells. Certain proteins, such as AFP, intercellular adhesion molecule-1 (ICAM-1), IQ motif containing GTPase activating protein 2 (IQGAP2), claudin-1 (CLDN1) and tissue transglutaminase 2 (TGM2), were validated both in multiple cell lines and in 61 specimens of clinical HCC cases. TGM2 was overexpressed in some of the AFP-deficient HCC cells (SK-HEP-1 and Bel-7402) and in about half of the tumor tissues with low levels of serum AFP (17/32, AFP-negative HCC). Trace amounts of TGM2 were found to be expressed in the samples with high serum AFP (26/29, AFP-positive HCC). Moreover, TGM2 expression in liver tissues showed an inverse correlation with the level of serum AFP in HCC patients. Notably, TGM2 existed in the supernatant of the AFP-deficient SK-HEP-1, SMMC-7721 and HLE cells, and it was found to be induced in AFP-producing cells (HepG2) by specific siRNA silence assay. Serum TGM2 levels of 109 HCC patients and 42 healthy controls were further measured by an established ELISA assay; the levels were significantly higher in HCC patients, and they correlated with the histological grade and tumor size. These data suggest that TGM2 may serve as a novel histological/serologic candidate involved in HCC, especially for the individuals with normal serum AFP. These novel findings may provide important clues to identify new biomarkers of HCC and indirectly improve early detection of the disease.     

Isolation, Characterization, and Synthesis of AlphaFetoprotein from Neonatal Rat Brain

Monospecific anti-rat serum alpha-fetoprotein (AFP) IgG was coupled to cyanogen bromide-activated Sepharose-4B (4.5 mg/ml packed volume of gel) to yield an immunoaffinity matrix. The immunoaffinity column was used to isolate AFP from feto-neonatal rat brain. The purified AFP was immunologically and electrophoretically similar to serum AFP. It yielded a single band with a molecular weight of 70,000 on sodium dodecyl sulphate polyacrylamide gel electrophoresis. Polyacrylamide gel electrophoresis of the protein under nondenaturing conditions yielded two charge variants of AFP, reminiscent of AFP from feto-neonatal rat serum. The AFP was observed to bind estradiol with Ka = 5.8 X 10(8) M -1 and 1.3 X 10(8) M -1 by dextran-coated charcoal adsorption and Sephadex gel filtration techniques, respectively. Newborn rat brain cells linearly incorporated [14C]leucine into immunoprecipitable AFP during 6 h in culture. It is, therefore, concluded that feto-neonatal rat brain contains AFP similar to that present in fetal serum and that it may arise in brain as a result of its in situ synthesis.     

Thyroxine-induced changes in the glycosylation pattern and in brain and serum levels of rat alpha-fetoprotein

We have studied the effect of thyroid disfunction during the postnatal period, on the serum and brain levels of rat alpha-fetoprotein (AFP) and albumin. Hypothyroidism was induced by treatment of pregnant rats and their newborn pups with 2-mercapto-1-methylimidazole(methimazole). Hyperthyroidism was provoked in newborns by daily injections of thyroxine (0.25 micrograms/g body wt) from the 3rd postnatal day weaning. Impaired growth, lower brain size, altered behaviour and morphological features observed were according to an altered thyroid status. Hypothyroid rats showed a significantly reduction in serum AFP concentration (78% of control values at 8 days of age) and a slight increase in that of albumin. level could be appreciated. Thyroxine supplementation (0.2 micrograms/rat/day) corrected most of these alterations. Hyperthyroidism induced a drastic fall in both serum and brain AFP levels (about 48% of the corresponding control values). Albumin concentration in serum was augmented significantly from the 12th postnatal day, but its brain levels did not change significantly. In hyperthyroid rats, a significant reduction (37% relative to controls) in the concanavalin A-non reactive microform of AFP, was observed. This alteration of the glycosylation pattern of AFP could be due to the inhibition by thyroxine of the activity of the hepatic enzyme GlcNAc-transferase III.     

Zhi-mu saponin inhibits alpha-fetoprotein gene expression in developing rat liver

1. A saponin isolated from the Chinese herb zhi-mu (Anemarrhena asphodeloides Bunge) modifies alpha-fetoprotein production when injected into newborn rats. 2. The serum level of AFP was determined quantitatively by immunorocket electrophoresis. 3. AFP serum levels were reduced to 60% of the control by zhi-mu saponin (ZMS). 4. The lower AFP level in drug treated rat serum is not due to a change in the pattern of serum AFP variants. 5. AFP mRNA levels in ZMS-treated rat livers, measured by RNA dot hybridization, decreased to about 50% of control levels after 4 days treatment. 6. Results from tritium labeled dexamethasone competition assays suggest that ZMS may act on AFP gene expression through glucocorticoid receptor mediated action.     

Strain and sex differences in serum alpha-fetoprotein levels in Mus musculus

Normal levels of circulating serum alpha-fetoprotein (AFP) in different inbred strains of mice, as determined by radioimmunoassay, are reported. These strains show different and characteristic values for the mean serum AFP concentration, suggesting genetic control of the serum level of this protein. Furthermore, within each strain a difference in serum levels of the protein is observed between sexes; males have a significantly higher serum AFP concentration.     

磁共振扩散加权成像联合血清AFP、CA125、CEA、CA199检测在早期原发性肝癌中的诊断价值研究

摘要 目的：研究磁共振扩散加权成像（DWI）联合血清甲胎蛋白（AFP）、糖类抗原125（CA125）、癌胚抗原（CEA）、糖类抗原199（CA199）检测在早期原发性肝癌（PHC）中的诊断价值。方法：选取我院自2017年9月开始至2020年5月收治的63例早期PHC患者纳入研究,记作肝癌组,再取同期我院收治的61例良性肝病患者记作对照组。对所有受试者均实施DWI扫描,比较两组DWI图像信号强度。检测并比较两组血清AFP、CA125、CEA、CA199水平,以受试者工作特征（ROC）曲线分析上述各项血清学指标水平检测和DWI诊断早期PHC的效能。另外,比较PHC淋巴结转移患者和无淋巴结转移患者血清AFP、CA125、CEA、CA199水平。结果：肝癌组DWI信号强度为高信号人数占比高于对照组（均P<0.05）。肝癌组血清AFP、CA125、CEA、CA199水平均高于对照组（均P<0.05）。血清AFP、CA125、CEA、CA199水平联合DWI诊断早期PHC的曲线下面积、灵敏度以及特异度均高于上述各检查方式单独检测。PHC淋巴结转移患者的血清AFP、CA125、CEA、CA199水平均高于无淋巴结转移患者（均P<0.05）。结论：DWI联合血清AFP、CA125、CEA、CA199检测诊断早期PHC的价值较高,且淋巴结转移患者的血清AFP、CA125、CEA、CA199水平明显升高。     

Antifreeze protein gene transfer in Atlantic salmon.

Salmonids freeze to death if they come into contact with ice. Many marine fish species that inhabit icy sea waters synthesize antifreeze proteins (AFP) to protect them from freezing. Production of stable lines of freeze-resistant salmon and other species would greatly facilitate development of sea-pen aquaculture in many regions. We successfully introduced winter flounder AFP genes into Atlantic salmon. Research to date indicates stable genomic integration and low levels of expression of winter flounder AFP genes in a small number (approximately 3%) of salmon developed from microinjected eggs. Inheritance of the AFP gene by offspring (F1) from crosses between transgenic and wild-type salmon revealed that the transgenic flounders (F0) were germ-line mosaics. Low levels of AFP precursors could be detected in the blood of all these transgenic offspring (F1). Approximately 50% of the progeny produced by crosses between transgenic F1 and wild-types contained the AFP genes. These results demonstrate that stable germ-line transformed Atlantic salmon can be produced.     

Alpha-fetoprotein impairs APC function and induces their apoptosis

alpha-Fetoprotein (AFP) is a tumor-associated Ag, and its serum level is elevated in patients with hepatocellular carcinoma (HCC). In vitro, AFP induces functional impairment of dendritic cells (DCs). This was demonstrated by the down-regulation of CD40 and CD86 molecules and the impairment of allostimulatory function. Also, AFP was found to induce significant apoptosis of DCs, and AFP-treated DCs produced low levels of IL-12 and TNF-alpha, a cytokine pattern that could hamper an efficient antitumor immune response. Ex vivo, APCs of patients with HCC and high levels of AFP produced lower levels of TNF-alpha than that of healthy individuals. In conclusion, these results illustrate that AFP induces dysfunction and apoptosis of APCs, thereby offering a mechanism by which HCC escapes immunological control.     

The role of cell interactions in the differentiation of teratocarcinoma-derived parietal and visceral endoderm

Cell interactions have been implicated in the differentiation of visceral and parietal endoderm in the developing mouse embryo. Embryoid bodies formed from F9 embryonal carcinoma cells have been useful in characterizing the events which lead to endoderm formation. As part of our effort to specify the interactions which may be involved in this process we have isolated visceral endoderm-like cells (VE) from F9 embryoid bodies and cultured them under various conditions. Using a combination of immunoprecipitation and enzyme-linked immunosorbent assay, we demonstrate that monolayer culture of these cells on a number of different substrates leads to a dramatic decrease in the level of alphafetoprotein (AFP), a VE-specific marker. Northern blot analysis of AFP mRNA indicates very low levels of this message are present after 48 hr in monolayer culture. Coincident with the drop in AFP levels is an increase in the levels of the cytokeratin Endo C and tissue plasminogen activator, both markers for parietal endoderm (PE). Morphological evidence at the ultrastructural level supports a transition from VE to PE. In contrast, the VE phenotype can be maintained in vitro by interaction with aggregates, but not monolayers, of stem cells. In addition, culturing the cells on the curved surface of gelatin-coated dextran beads, but not on a flat gelatin surface facilitates AFP expression and the cells are morphologically intermediate between VE and PE cells. The potential role of junctional complexes and cell shape are discussed.     

Synthesis of alpha-fetoprotein by the pre-implantation and post-implantation bovine embryo

The synthesis of alpha 1-fetoprotein (AFP) was measured by radioimmunoassay in tissues and fluids of 19 bovine embryos (14-46 days of gestation) and in tissue cultures of 4 pre-implantation embryos (17-27 days) by incorporation of radioactive methionine. AFP was first detected in Day-14 trophoblasts and secretion of AFP into allantoic fluid occurred by Day 16. Embryonic tissues and fluids in pre-implantation and post-implantation embryos contained levels of AFP that were 550 to 1 500 000 times higher than those found in maternal serum (3.9-298 000 compared with 0.07-0.25 ng/mg protein). High levels of AFP were also found in uterine fluid which suggested significant transfer of this protein from the early post-implantation conceptus. The major sites of AFP synthesis were yolk sac and fetal liver. It is concluded that the synthesis of bovine AFP is not initiated by events associated with implantation.     

血清miR-21 及AFP 在大鼠肝癌发生过程中的动态表达比较分析

目的:分析血清miR-21在肝癌发生过程中的表达水平并将其与传统肝癌血清标志物甲胎蛋白(AFP)比较,探索其成为肝癌早期诊断血清标志物的可能性。方法:二乙基亚硝胺(DENA)腹腔注射诱导建立大鼠肝癌模型,建模过程中收集造癌各个阶段的血清。Realtime-PCR检测血清miR-21的表达情况,ELISA法检测血清AFP水平。结果:与正常组及纤维化期大鼠相比,miR-21在肝硬化期、肝癌早期、肝癌晚期的大鼠血清均有不同程度上调(P<0.05),AFP在肝癌早、晚期的大鼠血清明显上调(P<0.05);与肝硬化期大鼠比较,肝癌早、晚期大鼠体内的miR-21表达显著上调(P<0.05),AFP在肝癌早期、肝癌晚期均显著上调(P<0.05)。结论:血清miR-21参与了肝癌发生的过程,对于肝癌发生的各个阶段均有很大的指示作用,可能作为肝癌预防和早期诊断的一个潜在标志物。     

alpha-Fetoprotein values in monkey (Macaca mulatta) pregnancy.

The pregnant rhesus monkey's (Macaca mulatta) potential as a model for understanding the dynamics of alpha-fetoprotein (AFP) metabolism in human pregnancy was evaluated. AFP levels in maternal and fetal serum and amniotic fluid were determined by radioimmunoassay. Significant correlations were found between decreasing maternal serum, fetal serum and amniotic fluid AFP concentrations and increasing gestational age. However, these data are not consistent with the AFP changes reported in human pregnancy. It appears that this animal has limited applicability as a model in this aspect of human pregnancy.