- Email: [email protected]
Individual and combined usefulness of lipid associated sialic acid, mucoid proteins and hexoses as tumor markers in breast carcinoma
203
Cancer Letters, 51 (1990) 203-208
Elsevier Scientific Publishers Ireland Ltd.
Individual and combined usefulness of lipid associated sialic acid, mucoid proteins and hexoses as tumor markers in breast carcinoma P.S. Patel”, B.R. Baxi”, S.G. Adhvaryu” “Deportment
of Cancer
Ahmedabad-380
Biology
and *Department
and D.B. Balarb of Pathology,
The Gujarat
Cancer and Research Institute, Asarwa,
016 (India)
(Received 30 August 1989) (Revision received 13 February (Accepted 13 February 1990)
1990)
Summary Serum leuels of lipid associated sialic acid (LASA), mucoid proteins (MP) and hexoses (galactose + mannose) were measured in 41 breast cancer patients, 14 patients with benign breast diseases and 36 healthy age matched female individuals. In breast carcinoma patients, we have observed significant increase in the levels of the three markers compared with the controls (P < 0.001) and in MP and hexoses compared to the patients with benign breast diseases (P < 0.001). LASA and hexoses levels were significantly higher in benign breast diseases with respect to controls (P < 0.001 and P < 0.01, respectively). We evaluated the sensitivity and specificity of the markers individually and in combination. MP were most sensitive (71.8%) and specific (71.4%). Both sensitioity and specificity were increased when combinations of the markers were studied. Combination of MP with LASA was most sensitive while the (97.4%) combination of MP and hexoses was most specific (92.9%). LASA was significantly elevated in infiltrating duct carcinoma compared to lobular carcinoma {P < 0.001). MP and hexoses Correspondence to: P.S. Patel.
also showed higher mean value in infiltrating duct carcinoma than lobular carcinoma. The present study suggests that the combination of the markers inuestigated might be useful for diagnosis and classification of breast carcinoma.
Keywords: breast carcinoma; tumor markers; sialogfycoconjugates; lipid associated sialic acid; mucoid proteins; hexoses. Introduction The quest is ongoing for more reliable serum/plasma markers for detecting malignant diseases at an early stage and for evaluating various therapeutic approaches. Because of the crucial role of cell surface and membrane constituents in neoplastic behaviour, changes in normal serum glycoconjugates have long been associated with malignancies. Elevations of serum sialic acid as reflected by N-acetyl neuraminic acid have been observed in various types of cancers [21]. Serum/plasma sialic acid levels have been correlated with the stage of disease [14,17,18], tumor burden [10,22], degree of metastasis [14,15] and recurrence [14]. High sensitivity of lipid associated sialic
204 Table 1.
Clinical details of breast cancer patients.
Tumor stage
No. of patients
Age (< 45/> 45 years)
Site R/L breast
I
-
-
II IIla IIIb IV
6 18 9 8
2/4 10/8 6/3 3/5
acid (LASA) have been reported [4,11], but in further studies considerably lower sensitivity has been observed [6,X?]. Increased levels of serum hexoses (galactose + mannose) and mucoid proteins (MP) (Seromucoid) have been correlated with presence and extent of malignant diseases [ 1,241. Recent reports from our laboratory have revealed significant difference in the levels of glycoconjugates between benign and malignant conditions of lung [20] and haematologic disorders [ 191. The current investigation evaluates the relative usefulness (sensitivity and specificity) of serum levels of LASA, MP and hexoses for detecting breast carcinoma. Materials and methods
Patients (55) who were referred to the hospital as suspected cases of breast carcinoma and 36 age-matched individuals as controls were included in the present study. Clinical examination, radiological and mammographic findings and histopathological reports revealed 14 patients with benign breast diseases and 41 patients with breast carcinoma. The tumor staging of breast carcinoma was done as per UICC recommendations. No metastasis were noticed in with breast carcinoma at the time of diagnosis. Details of breast carcinoma patients have been summarized in Table 1. Prior to administration of any treatment venous blood samples of the patients were collected. Blood samples of the controls and
Lobular carcinoma
Infiltrating duct carcinoma
-
-
-
2/4 12/6 5/4 5/3
0 6 1 1
6 12 8 7
patients were collected between 1030 h and 2330 h. The sera were kept at - 20°C until analysed . Determination ofLASA For analysing LASA 50 ul of serum was treated with chloroform:methanol (2: 1 v/v). The lipid extract was partitioned with 0.5 ml cold distilled water. Aqueous phase containing the LASA fraction was precipitated with phosphotungstic acid. The precipitates were resuspended in 1 ml distilled water. LASA in the suspension was determined using resorcinol reagent [ 131. Estimation of MP and hexoses Separation of soluble perchloric acid and phosphotungstic acid precipitable fractions and measurement of hexoses were done according to the method of Winzlar [25] with minor necessary modifications. Protein contents of the (MP) fraction was determined according to Hartee [9]. Statistical methods The results were analysed statistically and values were expressed as mean f S.E. The level of significance was assessed by computing Student’s t-test. Only when the P-value was less than 0.01 the difference between the two groups was considered statistically significant. The i&a-assay and interassay coefficients of variation were between 3.6% and
Table 2.
Comparison
of levels of the markers in controls, Group
LASA (mg/dI) Mucoid Proteins (mg/dI) Hexoses (mg/dI)
Control Benign Malignant Control Benign Malignant Control Benign Malignant
benign and malignant breast diseases
Mean
f S.E.
Statistical significance
0.607 1.661 1.905 4.026 7.572 12.480 0.590 0.994 1.764
15.070 23.701 28.026 78.634 95.379 147.615 8.874 12.450 20.072
< 0.001’ NS” < 0.001** NS < 0.001 < 0.001 < 0.01 < 0.001 < 0.001
(P)
*
‘Comparison between controls and benign breast diseases. l ‘Comparison between benign and malignant breast diseases. l * ‘Comparison between controls and breast carcinoma. NS: Not significant.
7.5 % , respectively. Sensitivity was calculated as percentage of individuals with breast carcinoma who had levels of the markers above upper limit of the controls. Specificity was the percentage of individuals with benign breast diseases who had values of the markers within normal range [6]. When the utility of combination of any two markers was studied positivity was determined when both or either of the markers were positive. Results
be seen from the table, MP and hexoses had significantly higher levels (P < 0.001); whereas LASA levels, though elevated, were not statistically significant in breast carcinoma compared to benign breast diseases. Sensitivity and specificity of the markers are shown in Tables 3 and 4, respectively. Alterations in hexoses were comparatively less sensitive (59.0%) and alterations in LASA were comparatively less specific (42.9%) for breast carcinoma. Both sensitivity and specificity were higher when the markers were studied in combination with each other. MP in combination with LASA gave highly sensitive (97.4%) results. Combination of MP with hexoses revealed highly specific (92.9%) results for breast cancer detection.
The levels of LASA, MP and hexoses were as summarized in Table 2. It is evident from Table 2 that all the three biomarkers showed higher levels in patients in both benign and malignant diseases when compared with controls. Elevations in the three biomarkers were Table 3. statistically also significant in breast carcinoma as compared to the controls (P < 0.001). LASA and hexoses had significantly higher levels in benign conditions in comparison with controls (P< 0.001and P < 0.01, respecLASA tively), while elevations in MP were not statistiMucoid cally significant. proteins Table 2 also provides comparison between Hexoses benign and malignant breast diseases. As can
Individual and combined
sensitivity (%).
LASA
Mucoid proteins
Hexoses
69.2 97.4
97.4 71.8
94.9 82.1
94.9
82.1
59.0
206 Table4. Individual and combined
Discussion
specificity (46).
LASA
Mucoid proteins
Hexoses
LASA Mucoid
42.9 85.7
85.7 71.4
78.6 92.9
proteins Hexoses
85.7
92.9
64.3
Levels of the markers in lobular carcinoma and infiltrating duct carcinoma are compared in Fig. 1. Patients with infiltrating duct carcinoma showed higher mean values in comparison with lobular carcinoma. LASA levels were statistically also significant when compared between the two histologic types (P < 0.001). All the three markers were also compared between premenopausal and postmenopausal status of the patients. We have not observed any significant difference in the levels of these markers between the two groups
Among the substances that are shed from growing malignant cell gangliosides, in particular LASA showed significant elevations in serum from patients bearing cancer [4,13,16]. Thus, serum LASA has been proposed as useful tumor marker, more satisfactory than total sialic acid determination [5]. However, LASA is expected to behave as a non-specific tumor marker after shedding, and that is an important aspect of the turnover of normal cell surface constituents [Z] which occurs principally in both normal and malignant growing cells. The present investigation showed significantly elevated levels of LASA in breast carcinoma patients when compared with controls (P < 0.001). But, LASA from our results is less specific and sensitive for breast carcinoma. Both sensitivity and specificity were increased when LASA was combined with either MP or hexoses. Previous studies have also compared multiple tumor markers and have found that combination of multiple markers is more beneficial [4,8,23]. In the present report,
pco~ool r
30.0
2o.c
t
T <*o.o E"
10x
10.0
LASA Fig.1.Comparison mg/dl.
1, Controls;
Mucold
Proteins
Hexoses
of mean values of biomarkers in different histologic types bf breast carcinoma. 2, lobular carcinoma; 3, infiltrating duct carcinoma.
Results expressed
as
207
combination of LASA with MP was highly sensitive (97.4%)) while the combination of MP with hexoses was most specific (92.9 %) . The seromucoid and carbohydrate-rich fraction of proteins has been studied as an indicator of tumor presence and the changes in glycoproteins could be determined by their carbohydrate moities. From the data it is apparent that the levels of MP and hexoses in simultaneously are malignant patients increased with LASA levels, but MP and hexoses levels are determined more specifically than LASA in breast cancer patients (Table 2). Breadly et al. [3] have examined the possibilities of the source of increased protein bound in cancer and occurring carbohydrates reported that there may be glycoprotein synthesis by tumor itself. In more general studies these markers are found to be increased when compared with controls. Previous reports have also stated that the levels of the biomarkers bear a positive correlation in malignancies with stage of diseases [7,14,17,18]. We have observed significant difference in the levels of LASA and hexoses between stage II and stage IIIb. However, the number of individuals being less in each stage of the disease, results are not correlated with stage-wise disease activity. Infiltrating duct carcinomas are invasive than lobular carcinomas and, also, levels of different biochemical substances can be correlated with invasiveness of the disease. The present study revealed significantly higher levels of LASA (P< 0.001) and higher mean values of MP and hexoses in infiltrating duct carcinoma patients, compared to the lobular carcinoma patients. It might be possible to correlate levels of the markers with stage-wise disease activity and invasiveness of the disease after studying more number of patients. The present study thus suggests that the measurement of the markers be helpful in the diagnosis of breast carcinoma as well as in differentiating between lobular carcinoma and infiltrating duct carcinoma patients. The combined use of these markers provides high degree of marker positivity.
Acknowledgements The authors are grateful to Dr. T.B. Pate], Director, The Gujarat Cancer and Research Institute, Ahmedabad for providing necessary facilities, staff members of Department of Surgical Oncology for their co-operation and Directorate, Medical Education and Research, Gujarat State for their partial financial support. References 1
2 3
4
5
6
7
8
9 10
11
12
Adelbert, S.B., Michael, F.D., Philip, Z.S. and Frances, A.Q. (1974) Utility of serum protein bound neutral hexoses and t_.-fucose for estimation of malignant tumor extension evaluation of efficacy for therapy. Cancer Res., 34, 538-542. Black, P.H. (1983) Shedding from normal and cancer cellsurfaces. N. Engl. J. Med., 303, 1415-1416. Breadly, W .P., Blasco, A.P., Weiss, J.F., Alexander, Jc., Jr., Silverman, N A. and Cheretien, P.B. (1977) Correlations among serum protein bound carbohydrates, serum glycoproteins, lymphocyte reactivity and tumor burden in cancer patients. Cancer, 40,2264-2272. Dinistrian, A.M. and Schwartz, M.K. (1981) Plasma lipid bound sialic acid and carcinoembryonic antigen in cancer patients. Clin. Chem., 27, 1737-1739. Dinistrian, A.M., Schwartz, M.K., Katopodis, N., Fracchia, A.A. and Stock, C. (1982) Serum lipid bound sialic acid as a marker in breast cancer. Cancer, 50, 18151819. Erbil, K.M., Jones, J.D. and Klee, G.G. (1985) Use and limitations of serum total and lipid bound sialic acid concentrations as markers for colorectal cancer. Cancer, 55,404-409. Erbil, K.M., Sen, S.E., Zincke, H. and Jones, J.D. (1986) Significance of serum protein and lipid bound sialic acid as marker for genitourinary malignancies. Cancer, 57, 1389 - 1394. Gustafsson, H., Franzen, L., Grankvist, K., Anniko, M. and Henriksson, R. (1988) Glycoprotein tumor markers in head and neck neoplasms a consecutive study on CA-50. CA 19-9 and CEA. J. Cancer Res. Clin. Oncol., 114, 394 -398. Hartee, E.F. (1972) Determination of proteins - a modification of Lowry method. Anal. Biochem., 48,422-427. Henderson, M. and Kessel, D. (1977) Alterations in plasma sialyltransferase in patients with neoplastic disease. Cancer, 39, 1129-1134. Hirshaut, Y., Katopodis, N. and Stock, CC. (1981) Serum or plasma lipid sialic acid determinations in malignant disease. Proc. Am. Assoc. Cancer Res., 22, 739. Kakari, S., Michaelidis, G., Besbeas. S.T., Ferderigos. A.S., Tsitsoura, M. and Poulaki, E. (1984) Serum lipid bound sialic acid as a marker of gastrointestinal Cancer. Anticancer Res., 4 (Suppl l), 3-6.
13
14
Katopodis, N., Hirashaut, Y., Nancy, L.G. and Stock, CC. (1982) Lipid associated sialic acid test for the detection of human cancer. Cancer Res., 42,5270-5275. Kiricuta, O., Bojan, O., Comes, R. and Christian, R. (1979) Significance of serum fucose, sialic acid, hepatoglobin and phospholipid levels in the evolution and treatment of breast cancer. Arch. Geschwulst Forsch., 49, 106-112.
15
Lipton, A., Harvey, H.A., Delong, S., Allegra, J., White, D., Allegra, M. and Davidson, E.A. (1979) Glycoproteins and Human Cancer: I circulatory levels in cancer serum. Cancer, 43,1766-1771.
16
Moore, D.J., Hsu, H., Mayas, G., Kloppel, T., Classen, P. and Jacobson, W. (1981) Lipid associated sialic acid elevated in sera of carcinoma patients. Proc. Am. Assoc. Cancer Res., 22,22.
17
Moss, A. J., Bissada, N.K., Boyd, CM. and Hunter, W.C. (1979) Significance of protein bound neuraminic acid levels in patients with prostatic and bladder carcinoma. Urology, 13,182-184.
18
Mrochek, J.E., Dinsmore, R.S., Tormey, D.C. and Walkes, T.P. (1976) protein bound carbohydrates in breast cancer: Liquid chromatographic analysis for mannose, galactose, fucose and sialic acid in serum. Clin. Chem., 22, 1516-
1524.
19
20
21
22
23
Patel, P.S., Adhvayu, S.G. and Balar, D.B. (1988) Serum glycoconjugates in patients with anemia and myeloid leukemia. Tumori, 74,639-644. Patel, P.S., Baxi, B.R. and Balar, D.B. (1989) Significance of serum sialoglycoproteins in patients with lung cancer. Neoplasma, 36,53-59. Shamberge, R.J. (1984) Serum sialic acid in normals and cancerpatients. J. Clin. Chem. Clin. B&hem., 22,647651. Silver, H.K.B., Rangel, D.M. and Morton, D.L. (1978) Serum sialic acid elevations in malignant melanoma patients. Cancer, 41, 1497-1499. Vaiani, Q., Locatelli. E., Sensalari, G., Monti, M., Reggiani, S. and Cunietti, E. (1987) Plasma lipid-bound sialic acid and carcinoembryonic antigen in cancer patients. Individual and combined clinical usefulness J. Exp. Clin. Cancer, 6,195-198.
24
Walker,
C. and Gray,
B.N.
(1983)
proteins and carcinoembryonic Winzlar,
R.J.
Acute phase reactant
antigen
colon and rectum. Cancer, 52, 150-
25
as tumor markers.
in cancer
(1955) Determination of serum glycoproof biochemical analysis, Vol. II, pp. 291
teins. In: Methods -311.
of the
154.
Editor: D. Glick. New York-London.
Cancer Letters, 51 (1990) 203-208
Elsevier Scientific Publishers Ireland Ltd.
Individual and combined usefulness of lipid associated sialic acid, mucoid proteins and hexoses as tumor markers in breast carcinoma P.S. Patel”, B.R. Baxi”, S.G. Adhvaryu” “Deportment
of Cancer
Ahmedabad-380
Biology
and *Department
and D.B. Balarb of Pathology,
The Gujarat
Cancer and Research Institute, Asarwa,
016 (India)
(Received 30 August 1989) (Revision received 13 February (Accepted 13 February 1990)
1990)
Summary Serum leuels of lipid associated sialic acid (LASA), mucoid proteins (MP) and hexoses (galactose + mannose) were measured in 41 breast cancer patients, 14 patients with benign breast diseases and 36 healthy age matched female individuals. In breast carcinoma patients, we have observed significant increase in the levels of the three markers compared with the controls (P < 0.001) and in MP and hexoses compared to the patients with benign breast diseases (P < 0.001). LASA and hexoses levels were significantly higher in benign breast diseases with respect to controls (P < 0.001 and P < 0.01, respectively). We evaluated the sensitivity and specificity of the markers individually and in combination. MP were most sensitive (71.8%) and specific (71.4%). Both sensitioity and specificity were increased when combinations of the markers were studied. Combination of MP with LASA was most sensitive while the (97.4%) combination of MP and hexoses was most specific (92.9%). LASA was significantly elevated in infiltrating duct carcinoma compared to lobular carcinoma {P < 0.001). MP and hexoses Correspondence to: P.S. Patel.
also showed higher mean value in infiltrating duct carcinoma than lobular carcinoma. The present study suggests that the combination of the markers inuestigated might be useful for diagnosis and classification of breast carcinoma.
Keywords: breast carcinoma; tumor markers; sialogfycoconjugates; lipid associated sialic acid; mucoid proteins; hexoses. Introduction The quest is ongoing for more reliable serum/plasma markers for detecting malignant diseases at an early stage and for evaluating various therapeutic approaches. Because of the crucial role of cell surface and membrane constituents in neoplastic behaviour, changes in normal serum glycoconjugates have long been associated with malignancies. Elevations of serum sialic acid as reflected by N-acetyl neuraminic acid have been observed in various types of cancers [21]. Serum/plasma sialic acid levels have been correlated with the stage of disease [14,17,18], tumor burden [10,22], degree of metastasis [14,15] and recurrence [14]. High sensitivity of lipid associated sialic
204 Table 1.
Clinical details of breast cancer patients.
Tumor stage
No. of patients
Age (< 45/> 45 years)
Site R/L breast
I
-
-
II IIla IIIb IV
6 18 9 8
2/4 10/8 6/3 3/5
acid (LASA) have been reported [4,11], but in further studies considerably lower sensitivity has been observed [6,X?]. Increased levels of serum hexoses (galactose + mannose) and mucoid proteins (MP) (Seromucoid) have been correlated with presence and extent of malignant diseases [ 1,241. Recent reports from our laboratory have revealed significant difference in the levels of glycoconjugates between benign and malignant conditions of lung [20] and haematologic disorders [ 191. The current investigation evaluates the relative usefulness (sensitivity and specificity) of serum levels of LASA, MP and hexoses for detecting breast carcinoma. Materials and methods
Patients (55) who were referred to the hospital as suspected cases of breast carcinoma and 36 age-matched individuals as controls were included in the present study. Clinical examination, radiological and mammographic findings and histopathological reports revealed 14 patients with benign breast diseases and 41 patients with breast carcinoma. The tumor staging of breast carcinoma was done as per UICC recommendations. No metastasis were noticed in with breast carcinoma at the time of diagnosis. Details of breast carcinoma patients have been summarized in Table 1. Prior to administration of any treatment venous blood samples of the patients were collected. Blood samples of the controls and
Lobular carcinoma
Infiltrating duct carcinoma
-
-
-
2/4 12/6 5/4 5/3
0 6 1 1
6 12 8 7
patients were collected between 1030 h and 2330 h. The sera were kept at - 20°C until analysed . Determination ofLASA For analysing LASA 50 ul of serum was treated with chloroform:methanol (2: 1 v/v). The lipid extract was partitioned with 0.5 ml cold distilled water. Aqueous phase containing the LASA fraction was precipitated with phosphotungstic acid. The precipitates were resuspended in 1 ml distilled water. LASA in the suspension was determined using resorcinol reagent [ 131. Estimation of MP and hexoses Separation of soluble perchloric acid and phosphotungstic acid precipitable fractions and measurement of hexoses were done according to the method of Winzlar [25] with minor necessary modifications. Protein contents of the (MP) fraction was determined according to Hartee [9]. Statistical methods The results were analysed statistically and values were expressed as mean f S.E. The level of significance was assessed by computing Student’s t-test. Only when the P-value was less than 0.01 the difference between the two groups was considered statistically significant. The i&a-assay and interassay coefficients of variation were between 3.6% and
Table 2.
Comparison
of levels of the markers in controls, Group
LASA (mg/dI) Mucoid Proteins (mg/dI) Hexoses (mg/dI)
Control Benign Malignant Control Benign Malignant Control Benign Malignant
benign and malignant breast diseases
Mean
f S.E.
Statistical significance
0.607 1.661 1.905 4.026 7.572 12.480 0.590 0.994 1.764
15.070 23.701 28.026 78.634 95.379 147.615 8.874 12.450 20.072
< 0.001’ NS” < 0.001** NS < 0.001 < 0.001 < 0.01 < 0.001 < 0.001
(P)
*
‘Comparison between controls and benign breast diseases. l ‘Comparison between benign and malignant breast diseases. l * ‘Comparison between controls and breast carcinoma. NS: Not significant.
7.5 % , respectively. Sensitivity was calculated as percentage of individuals with breast carcinoma who had levels of the markers above upper limit of the controls. Specificity was the percentage of individuals with benign breast diseases who had values of the markers within normal range [6]. When the utility of combination of any two markers was studied positivity was determined when both or either of the markers were positive. Results
be seen from the table, MP and hexoses had significantly higher levels (P < 0.001); whereas LASA levels, though elevated, were not statistically significant in breast carcinoma compared to benign breast diseases. Sensitivity and specificity of the markers are shown in Tables 3 and 4, respectively. Alterations in hexoses were comparatively less sensitive (59.0%) and alterations in LASA were comparatively less specific (42.9%) for breast carcinoma. Both sensitivity and specificity were higher when the markers were studied in combination with each other. MP in combination with LASA gave highly sensitive (97.4%) results. Combination of MP with hexoses revealed highly specific (92.9%) results for breast cancer detection.
The levels of LASA, MP and hexoses were as summarized in Table 2. It is evident from Table 2 that all the three biomarkers showed higher levels in patients in both benign and malignant diseases when compared with controls. Elevations in the three biomarkers were Table 3. statistically also significant in breast carcinoma as compared to the controls (P < 0.001). LASA and hexoses had significantly higher levels in benign conditions in comparison with controls (P< 0.001and P < 0.01, respecLASA tively), while elevations in MP were not statistiMucoid cally significant. proteins Table 2 also provides comparison between Hexoses benign and malignant breast diseases. As can
Individual and combined
sensitivity (%).
LASA
Mucoid proteins
Hexoses
69.2 97.4
97.4 71.8
94.9 82.1
94.9
82.1
59.0
206 Table4. Individual and combined
Discussion
specificity (46).
LASA
Mucoid proteins
Hexoses
LASA Mucoid
42.9 85.7
85.7 71.4
78.6 92.9
proteins Hexoses
85.7
92.9
64.3
Levels of the markers in lobular carcinoma and infiltrating duct carcinoma are compared in Fig. 1. Patients with infiltrating duct carcinoma showed higher mean values in comparison with lobular carcinoma. LASA levels were statistically also significant when compared between the two histologic types (P < 0.001). All the three markers were also compared between premenopausal and postmenopausal status of the patients. We have not observed any significant difference in the levels of these markers between the two groups
Among the substances that are shed from growing malignant cell gangliosides, in particular LASA showed significant elevations in serum from patients bearing cancer [4,13,16]. Thus, serum LASA has been proposed as useful tumor marker, more satisfactory than total sialic acid determination [5]. However, LASA is expected to behave as a non-specific tumor marker after shedding, and that is an important aspect of the turnover of normal cell surface constituents [Z] which occurs principally in both normal and malignant growing cells. The present investigation showed significantly elevated levels of LASA in breast carcinoma patients when compared with controls (P < 0.001). But, LASA from our results is less specific and sensitive for breast carcinoma. Both sensitivity and specificity were increased when LASA was combined with either MP or hexoses. Previous studies have also compared multiple tumor markers and have found that combination of multiple markers is more beneficial [4,8,23]. In the present report,
pco~ool r
30.0
2o.c
t
T <*o.o E"
10x
10.0
LASA Fig.1.Comparison mg/dl.
1, Controls;
Mucold
Proteins
Hexoses
of mean values of biomarkers in different histologic types bf breast carcinoma. 2, lobular carcinoma; 3, infiltrating duct carcinoma.
Results expressed
as
207
combination of LASA with MP was highly sensitive (97.4%)) while the combination of MP with hexoses was most specific (92.9 %) . The seromucoid and carbohydrate-rich fraction of proteins has been studied as an indicator of tumor presence and the changes in glycoproteins could be determined by their carbohydrate moities. From the data it is apparent that the levels of MP and hexoses in simultaneously are malignant patients increased with LASA levels, but MP and hexoses levels are determined more specifically than LASA in breast cancer patients (Table 2). Breadly et al. [3] have examined the possibilities of the source of increased protein bound in cancer and occurring carbohydrates reported that there may be glycoprotein synthesis by tumor itself. In more general studies these markers are found to be increased when compared with controls. Previous reports have also stated that the levels of the biomarkers bear a positive correlation in malignancies with stage of diseases [7,14,17,18]. We have observed significant difference in the levels of LASA and hexoses between stage II and stage IIIb. However, the number of individuals being less in each stage of the disease, results are not correlated with stage-wise disease activity. Infiltrating duct carcinomas are invasive than lobular carcinomas and, also, levels of different biochemical substances can be correlated with invasiveness of the disease. The present study revealed significantly higher levels of LASA (P< 0.001) and higher mean values of MP and hexoses in infiltrating duct carcinoma patients, compared to the lobular carcinoma patients. It might be possible to correlate levels of the markers with stage-wise disease activity and invasiveness of the disease after studying more number of patients. The present study thus suggests that the measurement of the markers be helpful in the diagnosis of breast carcinoma as well as in differentiating between lobular carcinoma and infiltrating duct carcinoma patients. The combined use of these markers provides high degree of marker positivity.
Acknowledgements The authors are grateful to Dr. T.B. Pate], Director, The Gujarat Cancer and Research Institute, Ahmedabad for providing necessary facilities, staff members of Department of Surgical Oncology for their co-operation and Directorate, Medical Education and Research, Gujarat State for their partial financial support. References 1
2 3
4
5
6
7
8
9 10
11
12
Adelbert, S.B., Michael, F.D., Philip, Z.S. and Frances, A.Q. (1974) Utility of serum protein bound neutral hexoses and t_.-fucose for estimation of malignant tumor extension evaluation of efficacy for therapy. Cancer Res., 34, 538-542. Black, P.H. (1983) Shedding from normal and cancer cellsurfaces. N. Engl. J. Med., 303, 1415-1416. Breadly, W .P., Blasco, A.P., Weiss, J.F., Alexander, Jc., Jr., Silverman, N A. and Cheretien, P.B. (1977) Correlations among serum protein bound carbohydrates, serum glycoproteins, lymphocyte reactivity and tumor burden in cancer patients. Cancer, 40,2264-2272. Dinistrian, A.M. and Schwartz, M.K. (1981) Plasma lipid bound sialic acid and carcinoembryonic antigen in cancer patients. Clin. Chem., 27, 1737-1739. Dinistrian, A.M., Schwartz, M.K., Katopodis, N., Fracchia, A.A. and Stock, C. (1982) Serum lipid bound sialic acid as a marker in breast cancer. Cancer, 50, 18151819. Erbil, K.M., Jones, J.D. and Klee, G.G. (1985) Use and limitations of serum total and lipid bound sialic acid concentrations as markers for colorectal cancer. Cancer, 55,404-409. Erbil, K.M., Sen, S.E., Zincke, H. and Jones, J.D. (1986) Significance of serum protein and lipid bound sialic acid as marker for genitourinary malignancies. Cancer, 57, 1389 - 1394. Gustafsson, H., Franzen, L., Grankvist, K., Anniko, M. and Henriksson, R. (1988) Glycoprotein tumor markers in head and neck neoplasms a consecutive study on CA-50. CA 19-9 and CEA. J. Cancer Res. Clin. Oncol., 114, 394 -398. Hartee, E.F. (1972) Determination of proteins - a modification of Lowry method. Anal. Biochem., 48,422-427. Henderson, M. and Kessel, D. (1977) Alterations in plasma sialyltransferase in patients with neoplastic disease. Cancer, 39, 1129-1134. Hirshaut, Y., Katopodis, N. and Stock, CC. (1981) Serum or plasma lipid sialic acid determinations in malignant disease. Proc. Am. Assoc. Cancer Res., 22, 739. Kakari, S., Michaelidis, G., Besbeas. S.T., Ferderigos. A.S., Tsitsoura, M. and Poulaki, E. (1984) Serum lipid bound sialic acid as a marker of gastrointestinal Cancer. Anticancer Res., 4 (Suppl l), 3-6.
13
14
Katopodis, N., Hirashaut, Y., Nancy, L.G. and Stock, CC. (1982) Lipid associated sialic acid test for the detection of human cancer. Cancer Res., 42,5270-5275. Kiricuta, O., Bojan, O., Comes, R. and Christian, R. (1979) Significance of serum fucose, sialic acid, hepatoglobin and phospholipid levels in the evolution and treatment of breast cancer. Arch. Geschwulst Forsch., 49, 106-112.
15
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