5780447327

972

A. Edorh et al.

use for estimating a gene copy number. However, our results show clearly that digoxigenin-dUTP™ labelling provides the same results than ³²P labelling when considering accuracy and sensitivity. Such a non-radioactive method should be favoured sińce results are obtained morę rapidly, reagents may be conserved over months, and their manipulation does not reąuire specific installations.

REFERENCES

Bloom, H.J. and Richardson, W. W., Histological grading and prognosis in breast cancer: a study of 1,409 cases of which 359 have been followed for 15 years. Br. J. Cancer 1957,11: 359-377.

Borresen, A.L., Ottestad, L., Gaustad, A., Andersen, T.I., Heikkila, Jahnsen, T., Tveit, K.M. and Nesland J.M., Amplification and protein over-expression of the neu/ HER/c-erbB-2 protooncogene in human breast carcinomas: relationship to loss of gene sequences on chromosome 17, family history and prognosis. Br. J. Cancer 1990;62:585-590.

Dawkins, H.J.S., Robbins, P.D., Sama, M., Carrelo, S., Harvey, J.M. and Sterrett, G.F., c-erbB-2 amplification and overexpression in breast cancer: Evaluation and comparison of Southern biot, slot doi, ELISA and immunohistochemistry. Pathology 1993,25: 124-132. Edorh, A., Parache, R.M., Migeon, C., N’sossani, B. and Rihn, B., Expression de 1’oncoprotćine c-erbB-2 dans la maladie de Paget mammaire: etude immunohisto-chimique a 1’aide de trois anticorps. Pathol. Biol. 1995, 43: 584-589.

Ferre, F., Marchese, A., Pezzoli, P., Griffin, S., Buxton, E. and Boyer, V., Quantitative PCR: an overview. In: The Polymerase Chain Reaction. Mullis, K.B., Ferrć, F. and Gibbs, R.A. (eds.), Birkhauser, Boston, 1994, pp. 67-88. Gutman, M., Ravia, Y„ Assaf, D., Yamamoto, T., Rozin, R. and Shiloh, Y., Amplification of c-myc and c-erbB-2 protooncogenes in human solid tumors: Frequency and clinical significance. Int. J. Cancer 1989, 44: 802-805. Kems, B.J.M., Pence J.C., Huper, G., Kinney, R.B. and Iglehart, J.D., c-erbB-2 expression in breast cancer detected by immunoblotting and immunohistochemistry. J. Histochem. Cytochem. 1990,38: 1823-1830. Machotka, S.V., Garrett, C.T., Schwartz, A.M. and Callahan, R., Amplification of the proto-oncogenes int-2, c-erbB-2 and c-myc in human breast cancer. Clin. chini. Acta 1989,184:207-218.

Olsson, H., Borg, A., Fernó, M., Ranstam, J. and Sigurdsson, H., Her-2/neu anó int-2 proto-oncogene amplification in malignant breast tumors in relation to reproductive factors and exposure to exogcnous hormones. J. natn. Cancer Inst. 1991,83: 1484-1487.

Orlando, C., Sestini, R., Zentilin, L., Gelmini, S., Pinzani, P.,

Giacca, M. and Pazzagli, M., Image analysis in quantitative PCR. An application for the measurement of c-erbB-2 oncogene amplification in DNA from human tumors. J. Biolumin. Chemilumin. 1994, 9: 223-228.

Paterson, M.C., Dietrich, K.D., Danyluk, J., Paterson, A.H.G., Lees, A.W., Jamil, N., Hanson, J., Jenkins, H., Krause, B.E., McBlain, W.A., Slamon, D.J. and Foumey, R.M., Correlation between c-erbB-2 amplification and risk of recurrent disease in node-negative breast cancer .Cancer Res. 1991,51:556-567.

Rihn, B._t Bottin, M.C., Coulais, C. and Martinet, N., Evaluation of non-radioactive labelling of deoxy-ribonucleic acids. Part two: colorigenic methods and comparison withchemiluminescent methods. J. biochem. biophys. Meth. 1995a, 30: 103-112.

Rihn, B., Coulais,C., Bottin, M.C. and Martinet, N., Evaluation of non-radioactive labelling and detection of deoxyribonucleic acids. Part one: chemiluminescent methods. J. biochem. biophys. Methods 1995b, 30: 91-102.

Rihn, B., Coulais, C., Bottin, M.C., Zissu, D. and Edorh, A., Use of non-radioactive methods for the determination of transgene expression, sequence and copy-number. Celi. mol. Biol. 1995c, 41: 907-915.

Ro, J., El-Naggar, A., Ro, J.Y., Blick, M., Frye, D., Fraschini, G., Fritsche, H. and Hortobagyi, G., c-erbB-2 amplification in node-negative human breast cancer. Cancer Res. 1989; 49: 6941 -6944.

Schechter, A.L., Stern, D.F., Vaidyanathan, L., Decker, S.J., Drcbin, J.A., Greene, M.I. and Weinberg, R.A., The neu oncogene: an erbB-2 related gene encoding a 185,000-Mr tumor antigen. Naturę 1984, 312: 513-516.

Sestini, R., Orlando, C., Zentilin, L., Gelmini, S._t Pinzani, P., Bianchi, S., Selli, C., Giacca, M. and Pazzagli, M., Measuring c-erbB-2 oncogene amplification in fresh and paraffin-embedded tumors by competitive polymerase chain reaction. Clin. Chem. 1994, 40: 630-636.

Slamon, D.J., Clark, G.M.,Wong, S.G., Levin, W.J., Ullrich, A. and McGuire, W.L., Human breast cancer: correlation of relapse and survival with amplification of the HER-2/ neu oncogene. Science 1987, 235: 177-182.

Slamon, J.D., Godolphin, W., Jones, L.A., Holt, J.A., Wong, S.G., Keith, D.E., Levin, W.J., Stuart, S.G., Udove, J_M Ullrich, A., Press, M.F., Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 1989, 244: 707-712.

Tavassoli, M., Quirke, P., Farzaneh, F., Lock. N.J., Mayne, L.V. and Kirkham, N., c-erbB-2/c-erbA co-amplification indicative of lymph node metastasis, and c-myc amplification of high tumor grade, in human breast carcinoma. Br. J. Cancer 1989, 60: 505-510.

Tsuda, H., Hirohashi, S., Shimosato, Y., Hirota, T., Tsugane, S., Yamamoto, H., Miyajima, N., Toyoshima, K., Yamamoto, T., Yokota, J., Yoshida, T._t Sakamoto, H., Terada, M. and Sugimura, T., Correlation between long-term survival in breast cancer patients and amplification of two putative oncogene co-amplification units: hst-J/ int-2 and c-erbB-2/ear-l. Cancer Res. 1989, 49: 3104-

79