[CANCER RESEARCH 37, 2505-251 1, August 1977] Galactosyltransferase and Phosphatase Activities in Physiological Fluids from BALB/c Mice Bearing the YC8 Lymphoma1

Françoise Raimond-Pironneau, Nicole Achy-Sachot, Monique Bard, Maximilienne Turpin, and Evelyne Morel2

Institut dImmuno Biologie, Hópital Broussais, 96 rue Didot, 75674 Paris Cedex 14, France

SUMMARY Alterations of glycosyltransferase activity have been dem onstrated in human and animal cells (5, 6, 9, 17) and in Physiological fluids from adult BALB!c mice were col various tissue fluids (8, 10, 12, 14, 21) during different tumor lected weekly or daily after the i.p. injection of YC8 tumor processes. Particularly, soluble galactosyltransferase activ cells. ities have been found in some tissue fluids (8, 12, 21). More Galactosyltransferase activity on endogenous acceptors recently, a cancer-associated isoenzyme has been detected was enhanced from 1% in normal sera to 14.6% in sera in human sera (19, 22). collected on Day 11. Afterward , it decreased to 9.6%. In We have thought that ascitic tumor would be of great peritoneal fluids, galactosyltransferase activity was en interest: in ascitic fluids, soluble tumor antigens would be hanced from 0 to 45.3% on Day 16. In urines, no significant present in vivo and could be detected without extraction activity was detected. Galactose transfer was enhanced in procedures. In previous studies, by use of an exogenous sera and peritoneal fluids until Day 10, in the presence of glycoprotein acceptor, we have reported (15, 16) an en urines. Then, urines had an inhibitory effect. Variations of hanced galactosyltransferase activity in ascitic fluids and in optimal pH and apparent K,, for uridine diphosphate galac sera from YC8 lymphoma-bearing BALB!c mice. More re tose were observed among fluids collected from normal cently, we have been able to demonstrate modifications of and ascitic mice. galactosyltransferase activity on endogenous acceptors, by Phosphatase activity was 85.6% in normal sera, de action of urines from normal and tumor-bearing mice (2). creased until Day 10 (11.2%), and was enhanced once more In this paper, we have studied the variations of galactosyl (21.4%) on Day 13. In peritoneal fluids, phosphatase activity transferase activity on endogenous acceptors during the was low, except on Days 5 (12.4%) and 9 (11%). In urines, no tumor development in ascitic fluids, sera, and urines from activity was detected , except on Day 6 (5.6%). Urines had an YC8 lymphoma-bearing BALB/c mice. Moreover, we have inhibitory effect on sera and an activator effect on perito determined free galactose released from UDP-galactose by neal fluids. nucleotide pyrophosphatases and phosphatases. A decrease of phosphatase activity and an enhancement of galactose transfer were related to tumor growth, as well MATERIALS AND METHODS as to the appearance of effectors and/or acceptors in urines. UDP-['4C]galactose (274 mCi/mmole), [14C]galactose-1 -P (266 mCi/mmole), and [14C]galactose (48 mCi/mmole) were INTRODUCTION purchased from New England Nuclear, Boston Mass. Animals and Tumor. Adult male BALB/c mice were ob During recent years, it often has been demonstrated that tamed from the Centre National de Ia Recherche Scienti many tumors may be associated with appearance of new fique, Orleans-Ia-Source, France. YC8 cells were kindly glycoprotein and glycolipid antigens, or with enhancement supplied by Y. Crépinin our laboratory and transplanted of normal cell antigens which are usually undetectable. as an ascitic tumor at weekly intervals, by i.p. injections of 5 Furthermore, it has been assumed that the accumulation of x [email protected] kinds of experiments havebeenperformed: incomplete or abnormal glycoproteins and glycolipids 120 mice were used for weekly i.p. passages and 100 mice could be due to deficiency or changed specificity of were used for daily investigations. One hundred twenty some glycosyltransferases. The presence on cell mem BALB/c mice were kept as controls; among them, 100 were branes of nucleotide pyrophosphatases and phosphatases untreated and 20 were given an injection of 0.9% NaCI that may compete with glycosyltransferases for the same solution. Metabolic cages from MediLabo (Vaucresson, nucleotide substrates has also been discussed (7, 11) and France) were used for urine collection. recently demonstrated on lymphocyte cells (1). Sampling of Physiological Fluids. Tumor cell suspen

, This work was supported by Institut de Ia Sante et de Ia Recherche sions were harvested weekly or daily from the peritoneal Medicale, Unite 20, and by Centre National de Ia Recherche Scientifique, cavity and were further centrifuged at 700 x g at 4°to Laboratoire AssociéNo. 143. 2 To whom requests for reprints should be addressed. separate cells and ascitic fluids. Sera were collected at the Received December 15, 1976; accepted May 6, 1977. same time. A pool of 24-hr urines was collected just before

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the animals were killed. Punctures of the physiological significant variations are found in the urines. fluids were effected on control animals. Similar determinations have been made on fluids col Enzyme Assays. The complete assay system contained, lected daily during tumor growth. The data have confirmed except when otherwise indicated, in a final volume of 50 @d, those found at the 7th day. Protein and carbohydrate con the following constituents: physiological fluid (10 p.l), 0.1 M centrations decreased from 0 to 10 days in sera; conversely, Tris-HCI buffer at optimal pH (20 pi), 0.4 M MnCl2 (5 .d), and these concentrations were enhanced in peritoneal fluids UDP-[14C]galactose (5 /.Ll,16.8 nCi, 61 pmoles). throughout tumor growth. No significant variations have The incubation was carried out for 60 mm at 37°.The been found in urines. reaction wasstopped bytheaddition oflO piof0.1 M EDTA, Galactosyltransferase Activity in Physiological Fluids and the tubes were kept in ice. The identification of reaction from Normal and Ascitic Mice. In preliminary studies, we @ products was made on 2 aliquots of 20 using the chro have determined some requirements for galactose transfer matographic method previously described (14). A parallel from UDP-galactose to endogenous acceptors in various run with UDP-[14C]galactose, [14C]galactose-1-P, and [‘4C]- physiological fluids.4 Opti mal pH for galactosyltransferase galactose as markers was done. In this chromatographic activity was found to be pH 6.6, pH 6.4, and pH 8.0 for system, the radioactivity remaining at the origin represents peritoneal fluids, sera, and urines, respectively. the amount of [‘4C]galactose incorporated in the endoge In this report, we have studied galactosyltransferase ac nous acceptors; the phosphatase activity is given by the tivity in physiological fluids obtained from control animals amount of free [‘4C]galactose.This system does not allow and from tumor-bearing mice killed 7 days after i.p. cell such a good separation of [‘4C]galactose-1-P and UDP injections (Table 1). As demonstrated in previous papers (2, [14C]galactose, so we will not quantify the nucleotide-pyro 15), no galactose transfer can be detected, either in LN or in phosphatase activity. UN. In SN, a weak activity was found. In UA, no significant Protein and Carbohydrate Determination. Protein deter galactose transfer could be determined. Contrariwise, gal mination was made by the procedure of Lowry et a!. (13) actosyltransferase activity was enhanced more than 4-fold with bovine serum albumin as standard. Carbohydrate con and more than 8-fold in SA and LA, respectively, compared centrations were determined by the orcinol-sulfuric acid with SN. method [that of Tillmans and Philippi (20)]. On urines, 2 Influence of incubation time on galactosyltransferase ac subsequent precipitations were made, using 0.6 Nfinal per tivity showed a linear increase during the 1st 15 mm in the chioric acid and 5% phosphotungstic acid, according to the different physiological fluids (Chart 1). After 60 mm, galac method of Pironneau et a!. (18). The 2 precipitates were tose transfer was still enhancing in LA only. Values for UN pooled for protein and carbohydrate determinations. and LN were too low to be reported in this chart. Influence of protein concentration on galactosyltransfer ase activity is reported in Chart 2 for SN, SA, and LA. The RESULTS enzyme activity showed a linear increase with protein con centration in the 3 fluids. At the same protein concentration Protein and Carbohydrate Determinations in Physiolog the galactose transfer was 10 and 2 times higher in LA and ical Fluids. Means of 5 determinations of protein and carbo SA, respectively, than in SN. In UA, UN, and LN, values were hydrate contents have been made on pools of physiological too low to be reported here. Optimal pH's have been found fluids collected on control animals. Aesults obtained did to be 6.6 and 7.4 for LA and SA, and 6.4 and 7.6 for SN. In not show any differences between untreated controls and spite of the very low values found in UA, an optimal pH of animals given an injection of 0.9% NaCI solution. Therefore, 8.0 can be deterrñined(Chart 3). Enzymatic activity in LN in this report, we shall not differentiate between the 2 kinds and UN was too weak, and no significant variations were of control mice. Protein has been found to be: 53.4 ±3.5 detected during pH study. These results are in good agree mg/mi in SN,3 9.6 ±1.0 mg/mI in UN, and 4.5 ±2.0 mg/mI ment with those described elsewhere.4 in LN. The orcinol-sulfuric method has given the following The apparent Km'5 for UDP-gaiactose have been deter results: 3.48 ±0.35 mg/mI in SN, 3.50 ±0.40 mg/mI in UN, mined for SN, SA, and LA galactosyltransferases. They have and 0.29 ±0.15 mg/mI in LN. been found to be 1.7 x 10@M for SN enzyme and 2.3 x 10@ The same determinations were effected on pools of physi M for both SA and LA. ological fluids collected 7 days after i.p. tumor cell injec Phosphatase Activity versus UDP-Galactose In Physio tion. Protein was found to be 47.9 ±4.3 mg/mI in SA, 10.5 logical Fluids from Normal and Ascitic Mice. We have ±1 .0 mg/mI in UA and 35.4 ±1 .6 mg/mI in LA. The orcinol studied phosphatase activity in physiological fluids ob sulfuric acid method gave the following results: 2.80 ±0.30 tamed from control and tumor-bearing mice killed 7 days mg/mI in SA, 3.48 ±0.35 mg/mI in UA, and 1.33 ±0.17 mgI after i.p. cell injection (Table 1). Nearly 30% of total radioac ml in LA. As we have demonstrated previously (15), protein tivity was recovered as galactose in SN, and only 3% in SA. and carbohydrate concentrations are lower in sera from Conversely, a higher value for galactose counting was ascitic mice than in normal sera and are higher in peritoneal found in LA, compared with LN, but the too-low values fluids from ascitic mice than in normal peritoneal fluids. No should be considered carefully. No significant variations could be determined in urines. The time course of phosphatase activity showed a linear 3 The abbreviations used are: 5N, LN, and UN, sera, peritoneal fluids. and urines, respectively, from normal BALB/c mice; SA, LA, and UA, sera, perito neal fluids, and urines, respectively, all from BALB/c mice bearing the ascitic YC8 lymphoma. 4N.Achy-SachotandE.Morel,personalcommunication.

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1Enzymatic Table fluidsCompleteactivities versus UDP-galactose in physiological incubation mixture: 10 j.d of physiological fluid, 20 jxl of 0.1 MTris-HCI buffer (pH 6.6 for peritoneal fluids, pH 6.4 for sera, and pH 8.0 for urines), 5 @xIof 0.4 M MnCl2, and 5 @xIof UDP 7, [14C]galactose (16.8 nCi).Galactosyl

transferase Phosphatase activityaControlOrigin of fluids activitya mice 504 Peritoneal fluid 16 ±5@ 42 ±5C Serum 73 ±10 2065 ±220 5tYC8Urines 15 ±5b 32 ± lymphoma-bearingmice― Ascitic fluid 629 ±70 155 ±20 Serum 330 ±40 254 ±30 Urines 28 ±5b 58 ±1OC

a Results are expressed in cpm from 6.7 nCi UDP-[14C]galactose total. Mean ±S.D. for 5 determinations in duplicate (pools of 20 to 25 mice). b Insignificant values for galactosyltransferase activity. Blank was 20 ±10. 20 40 60 mg/mi Chart 2. Influence of protein concentration on galactosyltransferase ac C Insignificant values for phosphatase activity. Blank was 40 ± 20. tivity in sera (•)andperitoneal fluids ( 0) from YC8 lymphoma-bearing mice @1Fluids are harvested 7 days after i.p. tumor injection. and in sera from control animals (0). Incubation mixtures are those indicated in “MaterialsandMethods,―except for optimal pH's (6.4 for sera and 6.6 for E peritoneal fluids). Protein concentration of physiological fluid varies from 0 0. to 60 mg/mI. 0 E U

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@0 ..@ 9.0 pH ..—-@ -@---. — Chart 3. The pH dependence of galactosyltransferase activity in sera (•), 5- 60 120 mm peritonealfluids(0),andurines(U)fromYC8lymphoma-bearingmiceand in sera from control animals (0). Incubation mixtures are those indicated in Chart 1. Time course of galactosyltransferase activity in sera (•),perito “MaterialsandMethods,―except for pH's varying from 6.0 to 9.3. neal fluids ( 0), and urines (U) from YC8 lymphoma-bearing mice and in sera from control animals (0). Incubation mixtures are those indicated in “Materi als and Methods,―except for time varying from 0 to 120 mm and for optimal pHs (6.4 for sera, 6.6 for peritoneal fluids, and 8.0 for urines). peritoneal fluids, the galactose transfer was continuously enhancing during tumor growth from 0 to 45.3% of total increase for 30 mm in SN, 60 mm in SA, and 120 mm in LN radioactivity, especially since Day 11. In sera, the gaiactose and LA (Chart 4). transfer was maximal on Day 11 (14.6%) and decreased The variation of this activity, in relation to pH, has been slightly afterward (9.6%). determined in SN, SA, and LA (Chart 5). Two optimal pH's In previous experiments, we have studied the influence of can be demonstrated: 6.4 and 8.0 in SN and 6.8 and 8.2 in urines on galactosyltransferase activity in sera.4 In this pa SA. Phosphatase activity values that are too weak are found per, we have investigated the behavior of sera and pen in LA; these do not permit definition of the optimal pH. toneal fluids in the presence of unines collected on the Galactose Transfer and Tumor Development. Galacto same day (Charts 7 and 8). These assays have been made at syltransferase activities have been tested in the different pH 8.0, which has been demonstrated to be the optimal pH fluids daily during tumor development (Chart 6). An impor for galactose transfer in incubation mixtures containi@ng tant galactose transfer appeared in sera and in peritoneal urines and sera. In both experiments, unines seemed to fluids,whereasitslevelremainedvery low in urines.In have 2 kinds of effects on galactose transfer: during the 1st

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S 15 30 60 120 mm S 10 15 days Chart 4. Time course of phosphatase activity in sera (•)andperitoneal Chart6. Galactosyltransferaseactivityduring tumor growth in sera (•), fluids ( 0) from YC8 lymphoma-bearing mice, and in sera (0) and peritoneal peritoneal fluids (0), and urines (U) from YC8 lymphoma-bearing mice. fluids (J) from control animals. Incubation mixtures are those indicated in Incubation mixtures are those indicated in “Materialsand Methods,―except “MaterialsandMethods,―exceptfor times varying from 0 to 120 mm and for for optimal pH's (6.4 for sera, 6.6 for peritoneal fluids, and 8.0 for urines). optimal pH's (8.0 for sera and peritoneal fluids).

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6.0 7.0 5.0 9:0 Chart 5. The pH dependence of phosphatase activity in sera (•)and 5 10 15 aays peritoneal fluids ( 0) from YC8 lymphoma-bearing mice and in sera from Chart 7. Galactosyltransferase activity during tumor growth in sera (•), control animals (0). Incubation mixtures are those indicated in “Materials urines (U), and aliquots containing the 2 fluids (c) from YC8 lymphoma and Methods,―except for pH's varying from 6.0 to 9.3. bearing mice. Incubation mixtures are those indicated in “Materialsand Methods,―except for pH 8.0. stage of tumor growth, urines increased galactosyltransfer ase activity 2.2-fold in LA and 2.4-fold in SA; after 10 days, the tumor development, except on Day 6. At this time, a unines had an inhibitory effect. At the end of tumor growth, slight amount of free galactose (5.6%) was detected in when galactose transfer was 27.6% in LA and 5.4% in SA, chromatography. thisinhibitoryeffectwastotal. We have tested the effect of daily unines on phosphatase Phosphatase Activity and Tumor Development. The van activity in sera and penitoneal fluids. For these assays, we ation of phosphatase activity has been measured at pH 8.0 have used SA and LA collected 7 days after the tumor in the different fluids, daily during tumor growth (Chart 9). injection, in order to exclude specific variations due to This activity, which is very important in SN (85.6% of total these fluids (Chart 10). In the presence of unines, the activ radioactivity), strongly decreased in SA until Day 10 ity was strongly inhibited in sera. This effect was more sig (11.2%). After Day 10, its level was enhanced once more nificant on Days 3 and 8, and from Day 13 until death (21.4% on Day 13). In penitoneal fluids, the activity was (93% inhibition). On the other hand, the unines seemed to generally weak, except on Days 5 (12.4%) and 9 (11%). The have no inhibitory effect but had an activator effect on phosphatase activity may be neglected in unines during all penitoneal fluids phosphatase enzyme. Indeed, from Day 5,

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S 10 15 days Chart 8. Galactosyltransferase activity during tumor growth in peritoneal fluids ( 0), urines (U), and in aliquots containing the 2 fluids (•)fromYC8 lymphoma-bearing mice. Incubation mixtures are those indicated in “Materi als and Methods,―except for pH 8.0. -2

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K E Chart 10. Modification of phosphatase activity in fluids by urine from 0. YC8 lymphoma-bearing mice during tumor growth. The curves represent U the differences between galactose recovered in incubation mixtures con taming sera or peritoneal fluids plus urines and galactose recovered in incubation mixtures containing sera or peritoneal fluids. Sera and peritoneal fluids were collected 7 days after tumor cell injection; urines were collected daily. Incubation mixtures are those indicated in “MaterialsandMethods,― except for pH 8.0. Results are expressed in cpm of free galactose for [(SA + UA) —5AJ(0) and for [(LA + UA) —LA](•).

ceptors. In sera collected 7 days after i.p. injections of YC8 tumor cells, the specific activity of galactosyltransferase was 4-fold higher than in sera from normal BALB/c mice (1% oftotal radioactivity). This activity was at a maximum on Day 11 (14.6%) and decreased slowly until it reached a stationary level (9.6%). In penitoneal fluids, the amount of 2 galactose incorporation on endogenous acceptors was continuously enhancing during the tumor development un til the death of the animals (7.4% on Day 7 and 45.3% on Day 16). Kinetic studies on enzymatic activity have shown signif icant differences between physiological fluids from tumor beaning mice and those from control animals. These results can indicate either a modification of normal serum enzyme 5 10 15 aays or the appearance of a new one in fluids from tumor-bearing Chart 9. Phosphatase activity during tumor growth in sera (•),peritoneal mice. This appearance could be due, as has been sug fluids ( 0), and urines (U) from YC8 lymphoma-bearing mice. Incubation mixtures are those indicated in “MaterialsandMethods,―except for pH 8.0. gested (4, 15), to the leakage of tumor enzyme from either intact or lysed tumor cells into the circulation system of the the presence of unines intensified the enzymatic activity, host. In support of this view, a correlation would exist especially on Day 10 (9.5-fold). between tumor growth and fluid galactosyltransferase 1ev els. Results we have obtained in studying the galactose DISCUSSION transfer during tumor development are in good agreement with this hypothesis. Experiments made on senaand penitoneal fluids collected Previously, we failed to demonstrate the presence of daily during tumor growth have shown an important in galactosyltransferase activity on endogenous and exoge crease of galactosyltransfenase activity on endogenous ac nous acceptors in unines from control animals and from

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Downloaded from cancerres.aacrjournals.org on October 1, 2021. © 1977 American Association for Cancer Research. F. Raimond-Pironneau et a!. ascitic mice killed 7 days after i.p. tumor cell injection (2). In of the transfenase system; new enzymatic forms and/or new this report, no significant galactose transfer on endogenous acceptor molecules, relative to the phenomenon of tumori acceptors had been detected during tumor development. genesis, would appear. Furthermore, the previously postu These results did not permit us to determine whether the lated concept of tumor cell constituents release in host enzyme and/or the endogenous acceptors were lacking. circulation system cannot be excluded. Enzymatic assays Therefore, assays were performed with incubation mixtures on subcellulan fractionations of YC8 cells harvested at dif containing sera on penitoneal fluids and unines, in order to fenent stages of tumor development ane now under investi investigate the existence of endogenous acceptors in un gation, and these should permit us to favor one of these ines. In the presence of unines, the galactose transfer was hypotheses. enhanced in SA and LA until Day 10; then, unines had an inhibitory effect on galactosyltransferase activity in SA and LA. These results lead us to assume several hypotheses. REFERENCES First, acceptors for SA and LA galactosyltransferases could 1. Abney, E. A., Evans, W. H., and Parkhouse, A. M. E. Location of Nucleo be present in unines. Until Day 10, they would be added to tide Pyrophosphatase and Alkaline Phosphodiesterase Activities on the those of the SA and LA. Then they would be in excess and Lymphocyte Surface Membrane. Biochem. J., 159: 293-299, 1976. 2. Achy-Sachot, N., and Morel, E. Galactose Transfer and UDP-Galactose would compete, leading to an inhibitory process. Second, Hydrolysis in Urine from Normal BALB/c and BALB/c YC8 Mice. Ex the presence in unines of activators and inhibitors for SA perientia, 32: 1126-1128, 1976. and LA enzymes could be suggested, activation being 3. Bard, M., and Morel, E. Miseen Evidenced'ActivitésNuclöotide-Pyro phosphatase et Phosphohydrolase dans les Lymphocytes Normaux de stronger in the 1st step, and inhibition outpacing after Day Souris BALB/c et Absence de ces Activitósdans les Cellules Tumorales 10. The inhibitory process, which was already demonstrated YC8. Compt. Rend., 282: 1999-2002, 1976. in the unines from control animals (2), may or may not be 4. Bhattacharya, M., Chatterjee, 5. K., and Barlow, J. J. Uridine 5'-Diphos phate-Galactose: Glycoprotein Galactosyltransferase Activity in the specific for the tumor. Third, it may be assumed that SA and Ovarian Cancer Patient. Cancer Res., 36: 2096-2101 •1976. LA endogenous acceptors are used by a galactosyltransfer 5. Bosman, H. B. Sialyltransferase Activity in Normal and RNA- and DNA virus Transformed Cells Utilizing Desialyzed, Trypsinized Cell Plasma ase present in unines. This enzyme would be inhibited, in Membrane External Surface Glycoproteins as Exogenous Acceptors. the 2nd step, by degradation products that appeared in Biochem. Biophys. Res. Commun. 49: 1256-1262. 1972. unines during tumor development. Other experiments are 6. Caccam, J. F., and Eylar, E. H. Glycoprotein Biosynthesis: Purification and Characterization of a Glycoprotein-Galactosyltransferase from Ehr now in progress to elucidate these processes. lich Ascites Tumor Cell Membranes. Arch. Biochem. Biophys., 137, 315— In this report, besides the galactosyltransferase activity, 324, 1970. 7. Deppert, W., Werchau, H., and Walter, G. Differentiation between Intra we have investigated nucleotide pyrophosphatase and cellular and Cell Surface Glycosyltransferase Activity in Intact Cells and phosphatase activities. As opposed to the nucleotide pyro in Cell Homogenate. Proc. NatI. Acad. Sci. U. S., 71: 3068-3072, 1974. phosphatase activity, which cannot be easily measured, 8. Ebner, K. E., and McKenzie, L. M. Alpha-Lactalbumin and Galactosyl transferase in Rat Serum and Their Relationship to Milk Secretion. free galactose has been quantified with accuracy. Contrary Biochem. Biophys. Res. Commun., 49: 1624-1630, 1972. to the galactosyltransferase activity, phosphatase activity in 9. Fishman, P. H., McFarland, V. W., More, P. T., and Brady, A. 0. Ganglio sera diminished during tumor growth. Indeed, we have side Biosynthesis in Mouse Cells: Glycosyltransferase Activities in Nor mel and Virally Transformed Lines. Biochem. Biophys. Res. Commun., found a high phosphatase activity in SN, compared with 48: 48-57, 1972. that found in SA. Experiments made during tumor growth 10. Johnston, I. A., McGuire, E. J., and Roseman, S. Sialic Acids: A Urldine Diphosphate N-Acetylglucosamine Glycoprotemn N-Acetylglucosaminyl have shown an important decrease of the galactose forma transferase from Goat Colostrum. J. Biol. Chem., 248: 7281-7288, 1973. tion from Day 0 (85.6% of total radioactivity at pH 8.0) to Day 11. Keenan, T. W., and Morre, D. J. Glycosyltransferases: Do They Exist on 10 (11.2%). Until Day 10, no significant activity could be the Surface Membrane of Mammalian Cells? Federation European Bio chem. Soc. Letters, 55: 8-13, 1975. detected in penitoneal fluids. After 10 days, phosphatase 12. Kim, Y. S., Perdomo, J., and Whitehead, J. S. Glycosyltransferases in activity was enhanced in SA and LA. As for galactosyltrans Human Blood. Galactosyltransferase in Human 5erum and Erythrocyte ferase activity, we have found light variations between non Membranes. J. Clin. Invest.. 51: 2024-2039, 1972. 13. Lowry, 0. H., Rosebrough, N. J., Farr, A. L., and Randall, A. J. Protein mal and ascitic sera optimum pH's. These results are in Measurement with the Folin Phenol Reagent. [email protected]. , 193: 265- good agreement with preliminary experiments on YC8 cells 275, 1951. and on normal lymphocytes, which have demonstrated the 14. Mookerjea, S., Michaels, M. A., Hudgmn,A. L.. Moscarello, M. A. Chow. A., and Schachter, H. The Levels of Nucleotide-Sugar: Glycoprotemn block of phosphatase activity in tumor cells (3). This block Sialyl- and N-Acetylglucosaminyl-Transferases in Normal and Pathologi would exist until Day 10 in sera. Afterward, a new phospha cal Human Sera. Can. J. Biochem., 50: 738-740, 1972. 15. Morel, E., Achy-Sachot, N., Spik, G., and Montreuil, J. Characterization tase enzyme, related to the tumor, would appear in sera as of Glycoprotein-Galactosyltransferase Activity in Ascitic Fluids of BaIb/c well as in penitoneal fluids. Other kinetic studies are now in YC8 Mice. Federation European Blochem. Soc. Letters, 69: 171-174, progress to support this hypothesis. 1976. 16. Morel, E., Spik, G., and Montreuil, J. Mise en Evidence dune Activité Action of unines on phosphatase activity in SA and LA has Glycoprotéine-Galactosyltransférasedans les Fluide Ascitiques et les been shown to be different: phosphatase activity is inhibited Serums de Souris Porteuses de Ia Tumeur YC8. Compt. Rend., 282: 317- in SA and activated in LA. The activator could reveal, in 320, 1976. 17. Patt, L. M., and Grimes, W. J. Cell Surface Glycolipid and Glycoprotein ascitic fluids, a phosphatase activity otherwise undetecta Glycosyltransferases of Normal and Transformed Cells. J. BioI. Chem., ble. Further studies on this presumed activator-inhibitor 249: 4157-4165, 1974. 18. Pironneau, F., Sternberg, M., Moisy, M., Feret, J., Rebeyrotte, P., La system are now being undertaken. grue. G., and Jayle, M. F. Orosomucoid and Haptoglobin Excretion in All these data suggest that galactosyltransfenase and Rat Glomerulonephritis and Tubulonephritis. Nephron, 13: 434-442, phosphatase activities are related to tumor development. 1974. 19. Podolsky, D. K., and Weiser, M. M. Glycosyltransferase Activities in The degradation system existing in the control animals Human Sara: Detection of a Cancer-Associated Isoenzyme. Biochem. would be blocked in the tumor-beaning mice for the benefit Biophys. Res. Commun., 65: 545-551 , 1975.

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20. Tillmans, J., and Philippi, K. Uber den Gehalt den Wichtigen Protein der Galactose-Glycoprotein Galactosyltransferase of Rat Serum. Biochem. Nahrungsmittel an Kohlehydrat and @iberemKolorimetrisches Verfahren Biophys. Res. Commun., 45: 57-62, 1971. zUr Quantitativen Bestimmung von Stickstoffreiem Zucker in Eiweiss. 22. Weiser, M. M., Podolsky, D. K., and Isselbacher, K. J. Cancer-Associated Biochem. Z., 215: 36-60, 1929. Isoenzyme of Serum Galactosyltransferase. Proc. NatI. Aced. Sci. U.S., 21. Wagner, A. R., and Cynkin, M. A. Glycoprotein Metabolism: A UDP- 73: 1319-1322, 1976.

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Françoise Raimond-Pironneau, Nicole Achy-Sachot, Monique Barel, et al.

Cancer Res 1977;37:2505-2511.

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