Nucleotidase in Breast Cancer Patients1

Home , Fucosyltransferase, Galactosyltransferase, Sialyltransferase

[CANCER RESEARCH 38, 723-728, March 1978] Alterations in Serum Glycosyltransferases and 5 -Nucleotidase in Breast Cancer Patients1

Clement Ip and Thomas Dao

Department of Breast Surgery and Breast Cancer Research Unit, Roswell Park Memorial Institute, Buffalo, New York 14263

ABSTRACT isoenzyme was absent in control subjects, but it was signif icantly higher especially in patients with widespread metas We have measured sialyltransferase, galactosyltransferase, and fucosyltransferase as well as S'-nucleotidase tasis. In this report we have examined 3 glycosyltransferases in the serum of breast cancer patients. Serum sialyltrans (sialyltransferase, galactosyltransferase, and fucosyltrans ferase values in 65 normal healthy females ranged from ferase) as well as 5'-nucleotidase in the serum of breast 2.6 to 8.5 units, with a mean of 5.4. In 25 women with cancer patients with either potentially curable disease or operable primary breast cancer, serum sialyltransferase disseminated mÃ©tastases.Theobjective of this investigation levels were found to be between 6.2 and 15.4 units. was to determine whether the levels of these enzymes in Marked elevation of this enzyme level (range, 8.8 to 36 the serum can be correlated with the extent of the disease units) was observed in 48 patients with metastatic breast in these patients and to identify the most sensitive marker(s) cancer. Galactosyltransferase and fucosyltransferase for subsequent follow-up study in cancer patients. measurements, however, showed considerable overlap between the controls and the cancer patients. On the other hand serum 5 -nucleotidase and sialyltransferase MATERIALS AND METHODS in breast cancer patients showed very similar patterns. Materials. UDP-[fJ-'4C]galactose (specific activity, 274 Thus, serum 5 -nucleotidase values in 44 normal females mCi/mmole), CMP-[4,5,6,7,8,9-14C]sialic acid (specific ac ranged from 11.4 to 23.2 units, whereas the levels found tivity, 173 mCi/mmole), and [G-3H]AMP (specific activity, in 30 patients with metastasis were between 25 and 71.8 13.5 Ci/mmole) were purchased from New England Nu units. The tissue origin of abnormal levels of serum clear, Boston, Mass. GDP-[L/-'4C]fucose (specific activity, glycosyltransferases and 5 -nucleotidase was discussed 117 mCi/mmole) was bought from Amersham/Searle Corp., in relation to their physiological significance as well as Arlington Heights, III. Fetuin was purchased from Grand their role as markers for diagnosing early malignant Island Biological Co., Grand Island, N. Y. AMP, hen ovo- breast neoplasm and for monitoring the extent of metas mucoid (trypsin inhibitor; Type II-O), and A/-acetylneura- tasis. minic acid were obtained from Sigma Chemical Co., St. Louis, Mo. INTRODUCTION SF-fetuin3 for the sialyltransferase assay was prepared by mild acid hydrolysis of fetuin (27). SGF-fetuin for the galac Recently, there has been a growing interest in studying tosyltransferase assay was prepared by the periodate oxi- the alterations of circulating glycosyltransferases in cancer dation-borohydride reduction method described by Spiro patients. Kessel ef al. have reported elevated plasma sialyl (28). transferase (10, 11) and fucosyltransferase (12) in patients Serum Collection. Blood was collected from patients in with various types of malignant diseases. Kessel et al. (10, Vacutainers without anticoagulant. RBC were removed by 13) have also observed that changes in plasma sialyltrans centrifugation, and serum samples were stored at -70Â° ferase level were correlated with the course of the disease until ready for assay. in patients studied serially during therapy. Bhattacharya ef Enzyme Assays. For the determination of sialyltransfer al. (6) have reported an increase in serum galactosyltrans- ase, the assay mixture contained 50 fj.\ serum, 500 /Â¿gSF- ferase levels in patients with ovarian cancer. Similarly, fetuin, 1 nmole CMP-[14C]sialic acid (4.4 x 10s dpm), 1 ITIM fluctuations of this enzyme in the serum appeared to corre MnCU, and 20 m.M Tris-maleate buffer (pH 7.0) in a final late with tumor volume as well as the clinical status of the volume of 150 /J. For the determination of galactosyltrans patient. On the other hand Podolsky and Wieser (22) ferase, the assay mixture contained 20 /^l serum, 250 ^g showed that on the average the total serum galactosyltrans- SGF-fetuin, 3 nmoles UDP-['4C]galactose (2.2 x 10" dpm), ferase activity in cancer patients was only slightly higher 10 mM MnCU, and 20 HIM Tris-maleate buffer (pH 6.8) in a than that in normal" control subjects. However, by discon total volume of 100 ^1. For the assay of fucosyltransferase, tinuous polyacrylamide gel electrophoresis, Weiser ef al. the incubation reaction consisted of 40 Â¿Â¿Iserum,4 mg (31) detected an isoenzyme of galactosyltransferase in the ovomucoid, 2 nmoles GDP-['4C]fucose (5 x 10s dpm), 10 serum in 43 of 58 patients with various types of cancer. This mM MgCL, and 20 mivi Tris-maleate buffer (pH 6.8) in a total volume of 100 /Â¿I.For the determination of endogenous 1 This work was supported in part by Grant CA 14812-03, National Cancer activities in all 3 glycosyltransferase measurements, the Institute, NIH, USPHS, and by the Mary Plagier Cary Charitable Trust. ' To whom requests for reprints should be addressed, at Roswell Park Memorial Institute. 666 Elm Street, Buffalo, N. Y. 14263. 3The abbreviations used are: SF-fetuin, sialic acid-free fetuin; SGF, sialic Received August 8. 1977; accepted December 13, 1977. acid-free and galactose-free fetuin.

MARCH 1978 723

Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 1978 American Association for Cancer Research. C. Ip and T. Dao respective exogenous glycoprotein acceptor was omitted in RESULTS the reaction mixture. Tubes for the glycosyltransferase assays were incubated Table 1 summarizes some of the biological characteristics at 37Â°ina Dubnoff shaker for various periods of time: 1 hr of the serum sialyltransferase. Since the addition of a non- for sialyltransferase, 30 min for galactosyltransferase, and ionic detergent (Triton X-100) to the incubation mixture did 10 min for fucosyltransferase. These different incubation not affect the sensitivity of the enzyme measurement, it was times were chosen so that linearity of the reaction was therefore omitted in routine assays. achieved under the given assay conditions. The reaction The reaction catalyzed by sialyltransferase is stimulated was terminated by the addition of 2 ml ice-cold 5% trichlo- by Mn* ions and is specific for the transfer of sialic acid to roacetic acid:2% phosphotungstic acid in 0.5 N HCI, and the penultimate galactose residue. Native fetuin (which the tubes were centrifuged at 2,600 x g for 1C min. The contains terminal sialic acid residues) and SGF-fetuin are resultant pellet was washed twice with the same acid hence not suitable acceptors for the sialyltransferase assay. solution and once with ethanohdiethyl ether (2:1, v/v). The Fetuin, a fetal calf serum glycoprotein, has been shown to precipitate was dissolved in 1 ml NCS tissue solubulizer contain more than 1 oligosaccharide side chain with termi (Amersham/Searle) and placed in 10 ml scintillation cock nal sialic acid moieties (29). The sialic acid residue is tail consisting of Spectrafluor (Amersham/Searle):ethylene attached either Â«(2,3)to galactose or a(2,6) to /V-acetylga- glycol monoethyl ether:toluene (1:10:14, by volume) and lactosamine. The present assay system therefore conceiva 8% naphthalene. Radioactivity was determined by liquid bly measures the activity of 1 or more sialyltransferase(s), scintillation counting. All 3 glycosyltransferase activities although it is assumed that the majority of the sialic acid is were reported as exogenous activities calculated by the transferred to the galactose moiety in SF-fetuin. difference between total (in the presence of exogenous The endogenous activity (assayed in the absence of glycoprotein acceptor) and endogenous (in the absence of exogenous acceptor) is due to the transfer of sialic acid to exogenous acceptor) activities. The enzyme activities are glycoprotein substrates present in the serum. The endoge expressed as pmoles sugar transferred per mg protein per nous activities in serum samples containing high sialyl hr; 1 unit of glycosyltransferase activity is defined as 1 transferase values (from cancer patients) were moderately pmole sugar transferred per mg protein per hr. elevated but never exceeded 15% of the total enzyme For the assay of 5'-nucleotidase, the reaction mixture activity. Data reported in this paper concern only the exog contained 100 /J serum, 0.5 ^mole [3H]AMP (1.1 x 10s enous activities (total minus endogenous). dpm), 20 mM MgCI,, 50 mM Tris-HCI buffer (pH 8.5) in a Chart 1 illustrates the serum sialyltransferase activities in total volume of 800 /J. For exclusion of nonspecific phos- normal subjects, in women with benign breast lesions, and phatase, 0.5 Â¿Â¿mole2-glycerophosphatewas also added to in women with breast cancer. The control values obtained the medium as a "substrate diverter." Incubation was car in 65 healthy females between the ages of 18 and 74 ranged ried out at 37Â°ina Dubnoff shaker for 30 min. The reaction from 2.6 to 8.5 units with a mean of 5.4. The upper limit of the normal range was set arbitrarily at 8.2 units, i.e., 2 was stopped by the addition of 0.1 ml 0.25 M ZnSCs, S.D.'s from the mean. We observed no correlation between followed by 0.1 ml 0.25 M Ba(OH)2. Protein and unhydro- lyzed 5'-AMP were precipitated and removed by centrifuga- age and levels of serum sialyltransferase in this group of tion although adenosine remained in the supernatant. A normal subjects. 0.5-ml aliquot of the supernatant fraction containing Among 21 patients with benign lesions of the breast (all [3H]adenosine was then counted in 10 ml ACS (Amersham/ lesions were confirmed by biopsy), 12 (57%) had serum Searle). The results are expressed as nmoles 5'-AMP hydro- sialyltransferase levels that were within the normal range. lyzed per mg protein per hr; 1 unit of 5'-nucleotidase Serum sialyltransferase activities in 25 patients with primary activity is defined as 1 nmole of 5'-AMP hydrolyzed per mg breast cancer (Clinical Stages I and II) were measured protein per hr. before mastectomy. Although the serum sialyltransferase Separation of Radioactive CMP-Sialic Acid and Its Deg levels in some patients with cancer overlapped the control radation Product. Degradation of CMP-sialic acid by CMP- values, the enzyme levels in 17 (68%) extended beyond the sialic hydrolase (EC 3.1.4.40) in the serum during incuba tion was assessed by spotting 50 /Â¿Iofthe reaction mixture Table 1 on Whatman No. 3 paper and submitting it to chromatog- Characteristics of serum sialyltransferase activity raphy in the following solvent system: ethyl acetate:pyri- Enzyme activity dine:glacial acetic acid:water (5:5:1:3). Radioactive sialic Incubation system (dpm/mg protein/hr) acid bound to macromolecules remained near the origin, Complete" (total activity) 2056 whereas CMP-[14C]sialic acid donor and its degradation + 0.1% Triton X-100 2098 - Mn2' 1239 product [14C]sialic acid were separated into distinct peaks. - SF-fetuin (endogenous) A/-[14C]Acetylneuraminic acid (New England Nuclear) was 168 used as the standard. Exogenous acceptor specificity SF-fetuin'' Protein and Sialic Acid Determinations. Protein in serum 1888 was determined by the method of Lowry ef al. (18). For the Native fetuin 82 SGF-fetuin 148 determination of the total amount of bound sialic acid, serum samples were first hydrolyzed in 0.1 N H2SO4at 80Â° " The complete system is described in "Materials and Methods." '' The exogenous activity with SF-fetuin as the acceptor is for 1 hr. Sialic acid was then determined by the thiobarbi- calculated by subtracting the endogenous activity from the total turic acid method described by Aminoff (1). activity (2056 - 168 = 1888).

724 CANCER RESEARCH VOL. 38

Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 1978 American Association for Cancer Research. Serum Glycosyltransferases and Nucleotidase in Cancer upper limit of the normal range. The relationship between 16 the levels of serum sialyltransferase and the extent of lymph 15 node metastasis in these 25 primary cases is shown in Chart 2. Of the 14 cases with no lymph node metastasis, 11 5 14 patients (79%) showed sialyltransferase levels that were 13 higher than those in the normal range. Surprisingly, only 6 LJ g of 11 patients (55%) with 1 or more positive lymph nodes U- < 12 showed such an elevation in enzyme activities. The most l? noteworthy observation of this study was the finding that 48 ?i patients with systemic metastasis had significantly higher IO sialyltransferase levels in their serum than did controls in 9 their serum (Chart 1). i.s We had also evaluated the effect of increasing the con OLl * 8 centration of nucleotide-sugar donor to 200 nmoles by using radioactive CMP-sialic acid of lower specific activity 7 (0.96 mCi/mmole; New England Nuclear). In a study involv 6 ing 25 serum samples from control subjects, the sialyltrans ferase activities were increased 28- to 32-fold by increasing 5 the amount of CMP-sialic acid to 200 nmoles in the incuba r tion mixture. The same magnitude of increase was also 01234 II observed in 25 serum samples from cancer patients with POSITIVE LYMPH NODES Chart 2. Correlation between serum sialyltransferase activities and enzyme activities that were above the normal range. Thus, lymph node metastasis in primary breast cancer patients. although higher enzyme activities were obtained at a higher

substrate level, the relative values between controls and ~ 500 cancer patients were still maintained. For comparison of our results with those of other investigators (7, 10, 11,13, 14), data obtained with radioactive CMP-sialic acid of high- -35 specific activity are reported here. Measurement of CMP-sialic acid hydrolase in serum sam 400 â€¢J**ft;oAltCONTROL(SO)iX" -30 r ples from controls and cancer patients showed that 4 to 7% fu of substrate was degraded to sialic acid and that this activity *"p il was not related to the sialyltransferase level (results not 'â€¢Â°-25 9 - shown). It was therefore unlikely that changes in sialyltrans Â£â€¢Â§30Â°â€¢ Â«* ferase activity could be accounted for by excessive degra â€¢XX dation of the nucleotide-sugar donor. - 2O M Â»X* Chart 3 shows the serum galactosyltransferase and fucos- *1o Â¡ Â«Â« yltransferase activities. The upper limits of the normal range ZOO i1 s â€¢, i-0 -15 J for galactosyltransferase and fucosyltransferase were set as 284 and 209 units, respectively (2 S.D.'s above the mean). t00-BENIGN {:1 1*$ 5. - There was much overlap in these 2 enzyme activities be r$8IX4 - K> â€¢o01â€¢* tween the control and the cancer patients. In 24 serum too â€¢5

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PRIMARY METASTASIS DISEASECANCER(12) (15) (24) Chart 3. Serum sialyltransferase. galactosyltransferase, and fucosyl transferase activities in normal control subjects, women with benign breast lesions, and breast cancer patients. Numbers in parentheses, number of women.

i samples from patients with metastasis, both serum galac tosyltransferase and fucosyltransferase were found to be elevated above the upper limit of the normal range in only 14 cases (58%). The data clearly show that there is a lack of

CONTROL BENIGN PRIMARY ICTASTA3S correlation between these enzymes and the extent of the 1651 OSEASE CANCER IÂ«) (20 (29) disease in these patients. In contrast, the levels of sialyl Chart 1. Serum sialyltransferase activities in normal control subjects. transferase were found to be elevated above the normal women with benign breast disease, patients with primary breast cancer, and range in all 24 patients with metastasis. patients with systemic metastasis. Blood samples were drawn 2 to 3 days prior to mastectomy in primary breast cancer patients Points, 1 patient; The total amount of bound sialic acid in the serum was numbers in parentheses, number of women. determined, especially in patients with metastasis. Results

MARCH 1978 725

Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 1978 American Association for Cancer Research. C. Ip and T. Dao summarized in Table 2 show no change in serum sialic acid 25 content in breast cancer patients compared to that in the control subjects. No free sialic acid was detected in any serum sample. Ã• 20 5'-Nucleotidase, a marker enzyme for plasma membrane, was also examined in the serum of patients during this investigation. However, fewer cases were reported in this ÃŽ study: 17 patients with benign disease, 17 patients with primary breast cancer, and 30 patients with systemic metas tasis (Chart 4). The upper limit of the normal range in 44 Â«s , control females was set at 23.5 units (2 S.D.'s above the .1 io yh mean). All except 1 patient with benign disease showed li serum 5'-nucleotidase activities that were within the control I values. Of the 17 patients with primary breast cancer (6 of them had 1 or more positive lymph nodes), 11 (65%) showed activities above the upper limit of the normal range. The result was similar to that obtained with the sialyltransferase study. Among these 11 patients, 7 had no lymph node 20 40 60 80 SERUM S'-NUCLEOTIDASE metastasis, and 4 had axillary lymph nodes containing (nmolM S'-AMP hydrolyzid/mg protem/hr) metastasis. There appeared to be no discernible correlation Chart 5. Correlation between sialyltransferase activities and 5'-nucleo- between the extent of lymph node metastasis and the level tidase activities in the serum of breast cancer patients. O, patients with of this enzyme in the serum. However, 5'-nucleotidase primary breast cancer; â€¢patients with systemic metastasis. levels were distinctly higher in the 30 patients with systemic metastasis. galactosyltransferase assay in the present study measures A plot of sialyltransferase versus 5'-nucleotidase from the the enzymatic attachment of galactose into /3(1,4) linkage same serum sample in a total of 47 patients (17 patients with the A/-acetylglucosamine of the glycoprotein (EC with primary breast cancer and 30 patients with systemic 2.4.1.38). Changes in various glycolipid galactosyltransfer- metastasis) is shown in Chart 5. A positive correlation (r = ases have been reported in neoplastic cells. High activity of 0.80) was obtained between these 2 parameters. UDP-Gal:/ac Tri-Cer /3-galactosyltransferase (EC 2.4.1.86) was observed in cloned neuroblastoma cells (20) and in DISCUSSION guinea pig tumor cells (2). Moreover, this enzyme was 5- to 10-fold more active than were other galactosyltransferases This study shows that, of the 3 serum glycosyltransfer- such as UDP-Gal:GMI ÃŸ-galactosyltransferase (EC 2.4.1.62) ases examined, sialyltransferase is the most sensitive and UDP-Gal:/ac nTet-Cer a-galactosyltransferase (EC marker for measuring the extent of neoplastic disease in 2.4.1.87) (2). Thus, this study measures the fluctuation of these patients. Our results with galactosyltransferase (Chart only 1 of several galactosyltransferases that are known to 3) are comparable with the data described previously by be present in the mammalian system. Weiser ef a/. (31), who also used fetuin derivative as sub Recently, Kessel ef a/. (21) reported an elevation in strate. Weiser ef al. (31) have demonstrated that the mean fucosyltransferase activity in plasma of cancer patients. level of total serum galactosyltransferase from patients with Our results with fucosyltransferase (Chart 3) show that the carcinoma was only marginally higher than that in normal majority of breast cancer patients, even those with metas control subjects. The increase, however, was not statisti tasis, fail to manifest any discernible increase in this serum cally significant. The use of SGF-fetuin as the acceptor for enzyme activity. The fucosyltransferase assay performed in this study used ovomucoid as the glycoprotein acceptor, * eor as previously described by Reddy ef a/. (25), since we were unable to duplicate the assay system delineated by Chou ef a/. (9), with fetuin derivative as the acceptor. By the method of Reddy ef a/. (25), the rate of fucose transfer to ovomucoid was proportional to the time of incubation and the quantity of serum added. Because more than 1 fucosyltransferase exists in the serum (9, 21), the discrepancy between our I*Â» results and those reported by Kessel ef al. (12) could be due to the difference in substrate specificities of the fuco- 1 syltransferases examined in the 2 studies. While this work I0 was in progress, Bauer ef al. (3) have also reported that the a-3-fucosyltransferase level was elevated in the serum of cancer patients more than was that of a-2-fucosyltransfer- CONTROL BEMGN PfMMRY METASTASIS (44) DISEASE CANCER (SO) ase. On the other hand Khilanani ef a/. (15) found that 07) 07) plasma a-2-fucosyltransferase levels were correlated with Chart 4. Serum 5 -nucleotidase activities in normal control subjects, women with benign breast lesions, and breast cancer patients. Numbers in the percentage of marrow blast cells in patients with acute parentheses, number of women. myelogenous leukemias.

726 CANCER RESEARCH VOL. 38

Our results disclose that 43% of the patients with benign Table 2 breast disease had an elevation of their serum sialyltrans- Determination of total amount oÃbound sialic acid in serum ferase compared to 68% of those with primary breast of SubjectsNormal samples38 (/xg/m)S.E.650-900 Mean Â± cancer. The cause of the elevation of this serum enzyme control 760 Â±10.4 level in patients with benign breast lesions is not clear. The 660-940 771 Â± 9.5 seemingly inverse relationship between serum sialyltrans- Breast cancer patients 48Range with metastasisNo. ferase and lymph node metastasis (Chart 2) is an enigma at the present juncture. Further compilation of data and long- clearance rate (19). Another possibility is that the cell term follow-up of those patients with negative lymph nodes surface glycoconjugates have already been desialated be and high serum sialyltransferase are deemed necessary to fore they are released into the circulation. This hypothesis derive any relevant information on this aspect. is more plausible because neuraminidase has been shown Glycosyltransferases are primarily located in the Golgi to be elevated in malignant breast tumor (7). apparatus (26). However, recent biochemical (4, 30) and KlÃ¶ppel ef al. (17) have recently reported that serum electron microscopic (24) evidence shows that glycosyl- glycolipid-bound sialic acid content was elevated almost 2- transferases are also present on the cell surface. It has been fold in cancer patients. These increases, however, were not suggested that the abnormal levels of glycosyltransferases reflected by whole-serum sialic acid since the amount of in the circulation are due to "shedding" of plasma mem sialic acid in the ganglioside fraction constituted only a very brane materials from neoplastic cells (6, 10, 31). In the small percentage of the total. Preliminary results from our animal system, Kim ef al. (16) have demonstrated that laboratory concur with the finding of KlÃ¶ppelef al. (17). nonmetastasizing mammary tumors have a thick glycocalyx Our data showing increased levels of serum 5'-nucleotid- surface coat, whereas the spontaneously metastasizing ase in breast cancer patients represent a novel finding and tumors have little of this surface material. The absence of are in concert with the hypothesis that the abnormal levels the glycocalyx envelop has been attributed to increased of serum glycosyltransferases may originate from the tu shedding of plasma membrane components as evidenced mor. Since 5'-nucleotidase has been generally accepted as by the presence of cell surface antigen in the blood of rats a plasma membrane marker, the increase in this enzyme in bearing metastasizing tumors. Recently, Podolsky ef al. the circulation would presumably indicate accelerated (23) have shown that serum galactosyltransferase isoen- breakdown and release of cell surface material. Chatterjee zyme II, which is unique in tumor-bearing rats, has the ef al. (B) have shown that in plasma membrane-enriched same electrophoretic mobility as the enzyme derived from fraction derived from rat mammary carcinomas, the activi the tumor. Moreover, the activity of galactosyltransferase ties of 5 membrane marker enzymes (alkaline phosphatase, isoenzyme II is linearly related to the growth of the tumor. 5'-nucleotidase, phosphodÂ¡esterase I, nucleotide pyrophos- These authors have therefore suggested that isoenzyme II phatase, and alkaline RNase) were significantly lower in could have been released from the tumor cells into the the metastasizing tumor than they were in the nonmetastas circulation of the host. izing tumors. However, no measurement of these enzyme The physiological significance of these serum glycosyl activities in the serum was reported by the authors. transferases is far from being clear. Weiser ef al. (31) have The present study demonstrates that variations in 5'- shown that there is no consistent correlation between the nucleotidase activity correlate only with sialyltransferase activities of serum galactosyltransferase isoenzyme II and levels in the serum and do not correlate with the other 2 the carcinoembryonic antigen titers, suggesting that these glycosyltransferases, although galactosyltransferase has 2 parameters represent different chemical entities. The also been known to be present on the surface of neoplastic possibility that the serum glycosyltransferases could be cells (30). It is possible that sialyltransferase and 5'-nucleo- involved in the glycosylation of extracellular substrates tidase are intimately associated with each other on the seems rather remote due to the absence of nucleotide- plasma membrane and because of this proximity are sugars in the circulation. "sloughed off" from the cell surface together. The appear Recently, Bosmann and Hall (7) have reported that levels ance of tumor-specific isoenzymes of galactosyltransferase of sialyltransferase activity in human breast and colon and fucosyltransferase, which have different substrate malignant tissues were higher than those in adjacent "nor specificity, could explain the lack of correlation in the mal tissues." Elevated levels of cellular sialyltransferase variations of all 3 glycosyltransferases in the serum in a could conceivably lead to an enhancement in sialylation of consorted fashion, based on the premise that all originated glycoconjugates on the cell surface, which may then be from the cell surface. "released" into the circulation. Bernacki and Kim (5) Long-term follow-up study on serum sialyltransferase and showed that there was a direct relationship between in 5'-nucleotidase is currently being carried out in our labora creases in the amount of serum protein-bound sialic acid tory, with patients after mastectomy or endocrine ablation, and serum sialyltransferase activity in rats with transplant- to determine whether measurement of these 2 enzyme able metastasizing mammary tumors. In our study no ele markers will be of value in monitoring the appearance of vation in serum sialic acid content was detected in breast metastasis or remission of cancer after surgical resection cancer patients (Table 2). Conceivably, glycoconjugates and/or chemotherapeutic treatment. containing sialic acid residues from tumor plasma mem brane components are preferentially metabolized by the REFERENCES host. This seems rather unlikely because hypersialated 1. Aminoff. D. Methods for the Quantitative Estimation of N-Acetylneura- glycoprotein has been found to have a reduced plasma minic Acid and Their Application to Hydrolysate of Sialomucoids. Bio-

MARCH 1978 727

ehem. J..8ÃŽ: 384-392. 1961. Proc. Nati. Acad. Sei. U. S., 72: 1012-1016. 1975. 2. Basu, M., Basu, S.. Shanabruch, W. G., Moskel, J. R.. and Evans, C. H. 17. KlÃ¶ppel, T. M., Keenan, T. W., Freeman, M. J., and Morre, D. J. Lectin and Choiera Toxin Binding to Guinea Pig Tumor (104CI) Cell Glycolipid-Bound Sialic Acid in Serum: Increased Levels in Mice and Surfaces before and after Glycosphingolipid Incorporation. Biochem. Humans Bearing Mammary Carcinomas. Proc. Nati. Acad. Sci. U.S.. 74: Biophys. Res. Commun., 71: 385-392, 1976. 3011-3013, 1977. 3. Bauer, C., Kottgen, E.. and Reutter, W. Elevated Activities of a-2- and 18. Lowry, 0. H.. Rosebrough, N. J., Farr, A. L., and Randall. R. J. Protein Â«-3-Fucosyltransferases in Human Serum as a New Indicator of Malig Measurement with the Folin Phenol Reagent. J. Biol. Chem., 793: 265- nancy. Biochem. Biophys. Res. Commun., 76: 488-494, 1977. 275, 1951. 4. Bernacki, R. J. Plasma Membrane Ectoglycosyltransferase Activity of 19. Lunney. J.. and Ashwell, G. A Hepatic Receptor of Avian Origin Capable L1210 Murine Leukemic Cells. J. Cellular Physiol., 83: 457-466, 1974. of Binding Specifically Modified Glycoproteins. Proc. Nati. Acad. Sei. U. 5. Bernacki, R. J., and Kim, U. Concomitant Elevations in Serum Sialyl- S., 73: 341-343, 1976. transferase Activity and Sialic Acid Content in Rats with Metastasizing 20. Moskal, J. R., Gardner, D. A., and Basu, S. Changes in Glycohpid Mammary Tumors. Science. Ã•95: 577-580, 1977. Glycosyltransferases and Glutamate Decarboxylase and Their Relation 6. Bhattacharya, M., Chatterjee, S. K., and Barlow. J. J. Uridine 5'-Diphos- ship to Differentiation in Neuroblastoma Cells. Biochem. Biophys. Res. phate-Galactose:Glycoprotein Galactosyltransferase Activity in the Ovar Commun.,67: 751-758, 1974. ian Cancer Patient. Cancer Res., 36. 2096-2101, 1976. 21. Munro. J. R., and Schachter, H. The Presence of Two GDP-L- 7. Bosmann, H. B., and Hall, T. C. Enzyme Activity in Invasive Tumors of fucose:glycoprotein Fucosyltransferases in Human Serum. Arch. Bio Human Breast and Colon. Proc. Nati. Acad. Sei. U. S., 71: 1833-1837, chem. Biophys., 756: 534-542, 1973. 1974. 22. Podolsky, D. K., and Weiser, M. M. Galactosyltransferase Activities in 8. Chatterjee, S. K., Kim. U., and Bielat, K. Plasma Membrane Associated Human Sera: Detection of a Cancer-Associated Isoenzyme. Biochem. Enzymes of Mammary Tumors as the Biochemical Indicators of Metas Biophys. Res. Commun.. 65: 545-551, 1975. tasizing Capacity. Analysis of Enriched Plasma Membrane Preparations. 23. Podolsky, D. K., Weiser, M. M., Westwood, J. C., and Gammon, M. Brit. J. Cancer, 33. 15-26, 1976. Cancer-associated Serum Galactosyltransferase Activity. Demonstration 9. Chou, T. H., Murphy, C., and Kessel, D. Selective Inhibition of a Plasma in an Animal Model System. J. Biol. Chem., 252: 1807-1813, 1977. Fucosyltransferase by W-Ethylmaleimide. Biochem. Biophys. Res. Com 24. Porter, C. W., and Bernacki, R. J. Ultrastructural Evidence for Ectogly mun.. 74: 1001-1006, 1977. cosyltransferase Systems. Nature, 256 648-650, 1975. 10. Henderson, M., and Kessel, D. Alterations in Plasma Sialyltransf erase 25. Reddy. P. R. K., Tadolini, B., Wilson. J., and Williams-Ashman, H. G. Levels in Patients with Neoplastic Disease. Cancer, 39: 1129-1134, 1977. Glycoprotein Glycosyltransferases in Male Reproductive Organs and 11. Kessel, D., and Allen, J. Elevated Plasma Sialyltransferase in the Cancer Their Hormonal Regulation. Mol. Cellular Endocrinol., 5: 23-31, 1976. Patient. Cancer Res.. 35: 670-672, 1975. 26. Schachter, H.. Jabbal. !.. Hudgin, R. L., Pinteric, L., McGuire, E. J., and 12. Kessel, D., Chou, T. H., and Henderson, M. Determinants of Fucosyl Roseman, S. Intracellular Localization of Liver Sugar Nucleotide Glyco transferase Level in Plasma of the Cancer Patient. Proc. Am. Assoc. protein Glycosyltransferases in a Golgi-rich Fraction. J. Biol. Chem., Cancer Res., 17: 16. 1976. 245: 1090-1100. 1970. 13. Kessel. D., Samson. M. K., Shah, P., Allen, J., and Baker, L. H. 27. Spiro, R. G. Studies on Fetuin, a Glycoprotein of Fetal Serum. J. Biol. Alterations in Plasma Sialyltransferase Associated with Successful Chem., 235: 2860-2869, 1960. Chemotherapy of a Disseminated Tumor. Cancer, 38: 2132-2134, 1976. 28. Spiro. R. G. Periodate Oxidation of the Glycoprotein Fetuin. J. Biol. 14. Kessel, D.. Sykes. E., and Henderson, M. Glycosyltransferase Levels in Chem..239: 567-573, 1964. Tumors Metastatic to Liver and in Uninvolved Liver Tissue. J. Nati. 29. Spiro, R. G., and Bhoyroo, V. D. Structure of the O-Glycosidically Linked Cancer Inst., 59:29-32, 1977. Carbohydrate Units of Fetuin. J. Biol. Chem., 249: 5704-5717, 1974. 15. Khilanani, P., Chou, T-H., Lomen, P. L., and Kessel, D. Variation of 30. Verbert, A., Cacan, R.. and Montreuil, J. Ectogalactosyltransferase. Levels of Plasma Guanosine Diphosphate L-Fucose:/3-galactosyl a-2-u- Presence of Enzyme and Acceptors on the Rat Lymphocyte Cell Surface. Fucosyltransferase in Acute Adult Leukemia. Cancer Res., 37: 2557- European J. Biochem.. 70: 49-53, 1976. 2559, 1977. 31. Weiser, M. M., Podolsky, D. K., and Isselbacher, K. Cancer-Associated 16. Kim, U., Baumler, A., Carruthers, C., and Bielat, K. Immunological Isoenzyme of Serum Galactosyltransferase. Proc. Nati. Acad. Sei. U. S., Escape Mechanism in Spontaneously Metastasizing Mammary Tumors. 73: 1319-1322, 1976.

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Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 1978 American Association for Cancer Research. Alterations in Serum Glycosyltransferases and 5′-Nucleotidase in Breast Cancer Patients

Clement Ip and Thomas Dao

Cancer Res 1978;38:723-728.

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