Carcinoembryonic Antigen in Human Ovarian Neoplasms

[CANCER RESEARCH 35, 3807-3810,December 1975]

Anthony Marchand,1 Cecilia M. Fenoglio,2Robrt Pascal,3 Ralph M. Richart,4 and Sidney Bennett5

Departments of Pathology [A . M.. C. M. F., R. P., R. M. R.] and Surgery [S. B.], Columbia Presbyterian Medical Center. New York, New York 11X132

SUMMARY MATERIALS AND METHODS

The presence and location of cancinoembryonic antigen Antisera. Monospecific goat anti-CEA serum (Gol83) (CEA) was examined in tissue sections of 14 mucinous and and control serum, Co183 with the CEA specificity removed 13 serous cystadenomas and cystadenocancinomas of the by affinity chromatography, were kindly provided by Dr. F. ovary. CEA was demonstrated in the mucinous tumors but J. . Pnimus, Hoffman-La Roche, Inc., Nutley, N. J. The was not present in the serous tumors. In the mucinous antiserum (Go183) was tested by immunodiffusion and tumors, CEA was located in the glycocalyx and apical electrophoresis with homogenates of primary and meta portions of the absorptive-type epithelium, with only trace static colon carcinoma in order to verify specificity. It was quantities in goblet cells, a pattern identical to that seen in found to react with the unique site of CEA only (17). colonic neoplasms. Endocervical-type epithelium in the A rabbit anti-CEA serum (Rb 20) was also used. This mucinous tumors contained little or no CEA. antiserum was prepared by immunizing a rabbit with an extract of human colonic carcinoma followed by absorption INTRODUCTION with nonneoplastic tissues as previously described ( 15). This serum was found to contain antibody that recognized the Much attention has been focused on the antigenic proper antigenic site common to both colonic carcinoma antigen ties of various human cancers (3, 5, 19). Recent observa III and CEA but did not react with the colonic carcinoma tions have suggested that tumor-specific antigens may be antigen III specific antigenic site (17, 18). Preimmunization present in serous and mucinous cystadenomas of the ovary serum from the same rabbit was used as the control. (1, 2, 4, 6, 8, 13, 14). McNeil et al. (16) have suggested a Three-Layer Bridge Immunoperoxidase Procedure. The similarity in antigenicity between mucinous tumors of the 3-layer bridge immunoperoxidase procedure for antigen ovary and colonic tumors. Van Nagell et a!. (21) recently detection was performed according to the technique of Hsu suggested that ovarian cyst fluid CEA6 from mucinous et a!. ( I 1). The secondary and tertiary antisera for the goat tumors is immunochemically similar to colonic cancer CEA system were also supplied by Dr. Pnimus. In the rabbit on the basis of molecular size and immunoreactivity by system, the goat anti-rabbit globulin was purchased from immunodiffusion. Thus, it seemed relevant to investigate Cappel Laboratories, Downingtown, Pa. The rabbit the presence of CEA in mucinous tumors of the ovary and anti-horseradish penoxidase was prepared in this laboratory. to compare its location there to that of colonic tumors. In Following the reaction, sections were lightly countenstained addition, the localization of CEA in the various types of with nuclear fast ned. benign ovarian cystadenomas and cystadenocarcinomas was Tissues Examined. Fourteen mucinous tumors were ex studied. amined. Seven were adenomas, and 7 were carcinomas (Table 2). Thirteen serous tumors were studied, 5 benign and 8 malignant. The paraffin-embedded tissues were 1 Recipient of Special Fellowship Award 5ToICA-05107 from the obtained from the Division of Surgical Pathology and National Cancer Institute. Present address: Department of Pathology, Gynecologic Pathology. Control tissue specimens included 8 Hartford Hospital, Hartford, Conn. cases of colonic adenomas and adenocarcinomas, as well as â€˜Assistant Professor of Pathology and Associate Director of the Division of Obstetrical and Gynecologic Pathology, The Sloane Hospital sections ofnormal cervix (in various phases ofthe menstrual for Women. cycle), normal ovary with germinal (coelomic) inclusion 3 Associate Professor of Pathology and Associate Director of the cysts, and several products of conception. Central Tissue Facility of the Cancer Research Center (Center Grant The degree of staining was graded from I to 3, with CA-I3696-02 from the National Cancer Institute). minimal faint brown precipitate classified as Grade I. The 4 Professor of Pathology and Director of Pathology and Cytology Laboratories of the Obstetrical and Gynecological Service, the Sloane grading was evaluated according to the overall average Hospital for Women. To whom reprint requests should be addressed at, degree of staining. In cases with significant variation the 630 West 168th Street, New York, N. Y. 10032. range of grading was recorded. Cut edges of tissue sections S Research Associate, Department of Surgery, Cancer Research Center. and nuclear staining were ignored, because identical stain Supported by National Cancer Institute Grant 13696. aThe abbreviationusedis:CEA, carcinoembryonicantigen. ing was obtained with control serum and was considered to Received July 14, 1975; accepted September 9, 1975. be nonspecific (12).

DECEMBER 1975 3807

Downloaded from cancerres.aacrjournals.org on September 23, 2021. © 1975 American Association for Cancer Research. A . Marchand et al. RESULTS Table 2 Distribution of CEA in mucinous tumors Colonic Tumors. The location of CEA in the colonic locationGlycocalyxCytoplasmGobletCarcinomas92+1+1102+2+0Ii000122+(0-2+)'2+(0-2+)0Adenomas tumors is summarized in Table 1 and conforms to previous descriptions (9, 10). Identical results were obtained with the CaseCEA Gol83 and Rb 20 antisera. All colonic carcinomas exam med contained moderate to marked quantities (Grades 2 to 3) of CEA, located in the glycocalyx of tumor epithelium and the apical portions of the neoplastic absorptive cells. CEA in the colonic adenomas had an identical pattern, although the amount of CEA varied considerably from one (intestinalareas)132+(0-2+)2+(0-2+)0142+(0-2+)2+(02+)015I+(0-l+)l+(0-2+)0Adenomas area to another within the same tumor section (Fig. I). (Work currently in progress in our laboratories is designed to investigate the relationship between cellular atypia and expression of CEA in colonic adenomas.) An occasional (endocervicalareas)15'000160001700018000l9c000Atypical20d3+(2+--3+)e210002200(0-1+)0 neoplastic goblet cell contained intracellular traces of CEA. The surrounding normal mucosa in these cases contained no CEA (Fig. 1), except for 2 cases (Cases I and 2) in which there was slight patchy localization in the glycocalyx of normal glands immediately adjacent to the tumor. Mucinous Tumors. Table 2 contains a summary of the findings. Although the pattern ofCEA localization varies in this group, the intestinal-type epithelium of the mucinous tumors had a pattern ofCEA distribution that was identical to that of the colonic tumors (Fig. 2). Staining was only a Numbers in parentheses, range of reactivity. found in an occasional goblet cell. In only 1 carcinoma of â€˜Mixed intestinal and endocervical adenoma. C Mixed endocervical and serous adenoma. the intestinal type (Case 11) were we unable to detect CEA. d No gland formation. In contrast, the endocervical-type epithelium, whether aNo gobletcellsobserved. pant of a mixed endocervical-intestinal-type tumor or as the sole component of the tumor, had no demonstrable CEA composed purely ofgoblet cells. Since almost no CEA was (Fig. 3). As expected, I adenoma of a mixed endocervical found in goblet cells of the control colonic tissues, it is not serous type (Case 19) had no evidence of CEA. surprising that these 2 cases did not contain CEA. Three atypical mucinous tumors were also examined. In Serous Tumors. The serous cystadenomas and adenocar Case 20, a poorly differentiated ovarian carcinoma, there cinomas, as well as 1 case of a serous cystadenofibroma, were individual cells as well as small nests of cells widely contained no CEA. infiltrating the ovarian stroma, but neoplastic glands and, Control Tissues. With only 2 exceptions the control therefore, luminal surfaces of cells were absent. CEA was tissues contained no CEA. One exception occurred in a found in the cytoplasm of the tumor cells. In the 2 other specimen of endocenvical epithelium, biopsied during the atypical cases (Cases 21 and 22), the neoplasms were secretory phase of the menstrual cycle, that had a patchy distribution of CEA in the glycocalyx and glandular epithe Table I hal cytoplasm. We are unable to explain this isolated Distribution of CEA in colonic tumors finding. The other exception was a section of fetal adrenal and fetal kidney. Patchy staining was seen in the cytoplasm locationGlycocalyxCytoplasmGobletCarcinomasI3+(2+-3+r2+â€˜23+3+033+3+043+3+0Adenomas2c3+3+03c2+(I+-2+)2+(l+-2+)52+(0of both organs, but the amount of staining was identical to that seen in section incubated with the control sera. This CaseCEA may be due to the presence of peroxidase in these cells.

DISCUSSION The present investigation demonstrates the presence of CEA in mucinous tumors of the ovary. Previous studies from this laboratory indicate that a portion of the ovarian mucinous cystadenomas contain a population of intestinal like cells that resemble those present in colonic adenomas -2+)063+(0 -2+)2+(O (7). In addition, the ovarian mucinous cystadenocar -2+)073+(O -3+)2+(0 -3+)2+(l+-2+)083+(0 cinomas contain cells that resemble those found in colonic -3+)2+(0 -2+)0 carcinomas7 and, histochemically, there is a similarity

a Numbers in parentheses, range of reactivity. b No globlet cells seen in tissue sections. 7 C. M. Fenoglio, A. Ferenczy, and R. M. Richart. Mucinous Tumors of C Areas of adenomatous epithelium adjacent to carcinoma. the Ovary, 11. Ultrastructural Features of Mucinous Cystadenocar

3808 CANCER RESEARCH VOL. 35

Downloaded from cancerres.aacrjournals.org on September 23, 2021. © 1975 American Association for Cancer Research. CEA in Human Ovarian Neoplasms between the mucins secreted by the intestinal type of Urinary Bladder Tumors in Man. Intern. J. Cancer, 5: 39-46, 1970. ovarian tumors and colonic tumors.8 It is therefore, not 4. Chen, S., Koffler, D., and Cohen, C. J. Cell-Mediated Immunity in surprising that there is also a similarity in the CEA Patients with Ovarian Carcinoma. Am. J. Obstet. Gynecol., 115: distribution in the colonic tumors and the intestinal areas of 467-470,1973. the ovarian mucinous tumors. 5. Chu, E. H. Y., Stjernsward, J., Clifford, P., and Klein, G. Reactivity of Human Lymphocytes against Autochthonous and Allogenic Tu CEA is present only in the mucinous cystadenomas and mor Cells in Vitro. J. NatI. Cancer Inst., 39: 595-615, 1967. cystadenocarcinomas that contain intestinal-type epithe 6. DiSaia, P. J., Rutledge, F. N., Smith, J. P., and Sinkovics, J. G. hum. The presence of CEA in the apex of the neoplastic Cell-Mediated Immune Reaction to Two Gynecologic Malignant absorptive-type cells in these cases suggests intracellular Tumors. Cancer,28: 1129-1137,1971. antigen synthesis by the absorptive cell, followed by trans 7. Fenoglio, C. M., Ferenczy, A., and Richart, R. M. Mucinous Tumors port and incorporation of the antigen into the glycocalyx. of the Ovary. Ultrastructural Studies of Mucinous Cystadenomas This is consistent with what is known concerning glycopro with Histogenetic Considerations. Cancer, in press. tein synthesis (20). 8. Gall, S. A., Walling, J., and Pearl, J. Demonstration of Tumor The areas of the mucinous tumors, composed of endocer Associated Antigens in Human Gynecologic Malignancies. Am. J. Obstet. Gynecol., 115: 387-393, 1973. vical-type epithelium, contain virtually no CEA. This 9. Gold, P., Drupey, J., and Ansari, H. Position of Carcinoembryonic difference between intestinal and endocervical-type epithe Antigen of the Human Digestive System in Ultrastructure of Tumor hum may explain some of the conflicting results other Cell Surface. J. NatI. Cancer Inst., 45: 219-225, 1970. investigators have obtained in examining the antigenic 10. Gold, P., Gold, M., and Feedman, S. 0. Cellular Localization of properties of extracts of mucinous ovarian tumors. None of Carcinoembryonic Antigens of the Human Digestive System. Cancer these investigators separated the ovarian mucinous tumors Res.,28:1331-1334,1968. on the basis of the differentiation of the mucin-producing II. Hsu, K. C., Zimmerman, E. A., Rudin, L., Logerfo, P. Bennett, 5, epithelium into pure endocervical or mixed endocervical and Tannenbaum, M. Application of Immunoperoxidase Bridge intestinal type. Technique in Studies of Tumor Tissues with Antibody to Tumor As All the mucinous cystadenocarcinomas in this and our sociated Antigen (TAA). In: Proceedings of the Second Conference previously studied series have been of the intestinal type, and Workshop on Embryonic and Fetal Antigens in Cancer, 147-151. Oak Ridge. Tenn.: U. S. Atomic Energy Commission Division of Tech and the vast majority contain CEA. Endocervical-type nical Information, 1972. mucinous cystadenomas appear not to undergo malignant 12. Kataoka, K., Nakai, Y., and Fujita, H. The Fine Structural Localiza transformation or, if they do, to do so only rarely. If the tion of Peroxidase Activity in Digestive Organs of Rats and Mice. CEA can be detected in the blood or cyst fluid, its presence Histochemie, 38: 5-18, 1974. may be valuable in diagnosis or prognosis. 13. Levi, M. M., Keller, S., and Mandl, I. Antigenicity of Serous Cystadenocarcinoma Tissue Homogenates and Its Fractions. Am. J. Obstet. Gynecol., 105: 856-861, 1969. ACKNOWLEDGM ENTS 14. Levi, M. M., Parshley, M. S., and Mandl, I. Antigenicity of Papillary Serous Cystadenocarcinoma Tissue Culture Cells. Am. J. Obstet. The cooperation of Dr. Raffaele Lattes in allowing us to use paraffin Gynecol., 102: 433-439, 1968. blocks of colonic tumors is gratefully acknowledged. We also acknowledge 15. LoGerfo, P., Herter, F. P., LiVolsi, V., and Bennett, S. Studies on Dr. F. J. Primus who prepared many of the antisera used in this study. Tumor Associated Antigen: TAA. J. Surg. Res.. /5: 290, 1973. 16. McNeil, C., Ladle, N. J., Helmick, W. M., Trentelman, E., and Wentz, M. W. Antiserum to Ovarian Mucinous Cyst Fluid with Colon REFERENCES Cancer Specificity. Cancer Res., 29: 1539-1540, 1969. 17. Newman, E. S., Petras, S. E., Georgiadis, A., and Hansen, H. J. I. Bhattacharya, M., and Barlow, J. J. An Immunologic Comparison Interrelationship of Carcinoembryonic Antigen and Colon Carcinoma between Serous Cystadenocarcinoma of the Ovary and Other Human Antigen, III. Cancer Res., 34: 2125-2130, 1974. Gynecologic Tumors. Am. J. Obstet. Gynecol., 117: 849-853, 1973. 18. Newman, E. S., Petras, S. E., Hamilton, J. G., Hager, H. J., and 2. Bhattacharya, M., and Barlow, J. J. Immunologic Studies of Human Hansen, H. J. Demonstration of Two Tumor-Associated Antigens in Serous Cystadenocarcinoma of the ovary. Cancer, 3!: 588-595, 1973. Human Colonic Adenocarcinoma. Federation Proc., 3/: 639, 1972. 3. Bubenik, J., Perlmann, P., and Helmstein, K. Immune Response to 19. Sinkovics, J. G., Shirato, E., Gyorkey, F., Cabiness, J. R., and Howe, C. D. Relationship between Lymphoid Neoplasm and Immunologic Functions. In: 14th Annual Clinical Conference, University of Texas, p. 53. M. D. Anderson Hospital Year Book, Medical Publishers, 1970. 20. Spiro, R. G. Glycoproteins: Their Biochemistry, Biology, and Role in cinomas, submitted for publication to American Journal ofObstetrics and Gynecology. Human Disease. New EngI. J. Med., 28!: 991-1000, 1969. â€˜C.M. Fenoglio, G. A. Cottral, A. Ferenczy, and R. M. Richart. 21. Van Nagell, J. R., Pletsch, Q. A., and Goldenberg, D. M. A Study of Mucinous Tumors ofthe Ovary, III. Histochemical Studies, submitted for Cyst Fluid and Plasma Carcinoembryonic Antigen in Patients with publication to A merican Journal of Obstetrics and Gynecology. Cystic Ovarian Neoplasms. Cancer Res., 35: 1433- 1437, 1975.

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@ S 3b Fig. 1. Area of positive staining in colonic adenoma (top), and adjacent normal epithelium (bottom)stained for presence ofCEA. CEA is recognized by dense black precipitate on surface of adenomatous cells. x 200. Inset , higher magnification of such an area demonstrating reaction product on glycalyx and in apical cytoplasm. Normal tissue does not stain. x 500. Fig. 2. Mucinous cystadenocarcinoma of ovary. Neoplastic glands composed of intestinal-type epithelium, seen in cross-section, contain CEA on luminal surfaces. (x 1250). Fig. 3. Mixed intestinal endocervical-type mucinous cystadenoma. A, endocervical area stained for CEA contains no reaction product. Dark region at basilar portion of cells represents nuclei. B, intestinal area stained for CEA with reaction product at surface of cells. (A, x 700; B, x 1100).

Anthony Marchand, Cecilia M. Fenoglio, Robert Pascal, et al.

Cancer Res 1975;35:3807-3810.

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