Abstract
AIMS--To investigate the effects of fixation on the immunohistochemical demonstration of c-erbB-2 oncoprotein using paraffin wax and cryostat sections; to compare c-erbB-2 expression in non-neoplastic and neoplastic gastric tissues. METHODS--Adjacent blocks of tumour and non-neoplastic tissue from four gastrectomy specimens were put into a panel of 10 fixatives including acetone, B5, Bouin's fluid, Carnoy's fluid, buffered formalin, formol dichromate, zinc formalin, 4% paraformaldehyde, periodate-lysine-paraformaldehyde (PLP) and periodate-lysine-paraformaldehyde-dichromate (PLPD) before embedding in paraffin wax for sectioning. Similar tissue blocks were snap frozen and cryostat sections were postfixed in these fixatives, either alone or in combination, before immunostaining. RESULTS--In paraffin wax embedded sections the best fixative was PLP, and in frozen tissues the best results were obtained after fixation of cryostat sections in buffered formalin followed by cold methanol and acetone. Applying these fixatives to samples from a further 16 gastrectomy specimens, strong membrane staining of c-erbB-2 protein was found in the tumour in eight of 16 cases (50%) using paraffin wax sections, and staining was stronger in the better differentiated carcinomas. For frozen tissues, positive membrane staining was found in all gastric adenocarcinomas, but differential staining intensity associated with tumour differentiation could not be detected. CONCLUSIONS--These results indicate that fixation and paraffin wax embedding affect the results of immunohistochemical demonstration of c-erbB-2 in gastric cancer. The choice of fixative is critical in the demonstration and evaluation of c-erbB-2 protein expression by immunohistochemistry in gastric carcinomas. Staining results also vary depending on whether frozen or paraffin wax embedded tissues are studied.
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- Cordell J. L., Falini B., Erber W. N., Ghosh A. K., Abdulaziz Z., MacDonald S., Pulford K. A., Stein H., Mason D. Y. Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J Histochem Cytochem. 1984 Feb;32(2):219–229. doi: 10.1177/32.2.6198355. [DOI] [PubMed] [Google Scholar]
- Houldsworth J., Cordon-Cardo C., Ladanyi M., Kelsen D. P., Chaganti R. S. Gene amplification in gastric and esophageal adenocarcinomas. Cancer Res. 1990 Oct 1;50(19):6417–6422. [PubMed] [Google Scholar]
- Kameda T., Yasui W., Yoshida K., Tsujino T., Nakayama H., Ito M., Ito H., Tahara E. Expression of ERBB2 in human gastric carcinomas: relationship between p185ERBB2 expression and the gene amplification. Cancer Res. 1990 Dec 15;50(24):8002–8009. [PubMed] [Google Scholar]
- Lemoine N. R., Jain S., Silvestre F., Lopes C., Hughes C. M., McLelland E., Gullick W. J., Filipe M. I. Amplification and overexpression of the EGF receptor and c-erbB-2 proto-oncogenes in human stomach cancer. Br J Cancer. 1991 Jul;64(1):79–83. doi: 10.1038/bjc.1991.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Porter P. L., Garcia R., Moe R., Corwin D. J., Gown A. M. C-erbB-2 oncogene protein in in situ and invasive lobular breast neoplasia. Cancer. 1991 Jul 15;68(2):331–334. doi: 10.1002/1097-0142(19910715)68:2<331::aid-cncr2820680221>3.0.co;2-x. [DOI] [PubMed] [Google Scholar]
- Semba K., Kamata N., Toyoshima K., Yamamoto T. A v-erbB-related protooncogene, c-erbB-2, is distinct from the c-erbB-1/epidermal growth factor-receptor gene and is amplified in a human salivary gland adenocarcinoma. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6497–6501. doi: 10.1073/pnas.82.19.6497. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Slamon D. J., Clark G. M., Wong S. G., Levin W. J., Ullrich A., McGuire W. L. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science. 1987 Jan 9;235(4785):177–182. doi: 10.1126/science.3798106. [DOI] [PubMed] [Google Scholar]
- Slamon D. J., Godolphin W., Jones L. A., Holt J. A., Wong S. G., Keith D. E., Levin W. J., Stuart S. G., Udove J., Ullrich A. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science. 1989 May 12;244(4905):707–712. doi: 10.1126/science.2470152. [DOI] [PubMed] [Google Scholar]
- Tal M., Wetzler M., Josefberg Z., Deutch A., Gutman M., Assaf D., Kris R., Shiloh Y., Givol D., Schlessinger J. Sporadic amplification of the HER2/neu protooncogene in adenocarcinomas of various tissues. Cancer Res. 1988 Mar 15;48(6):1517–1520. [PubMed] [Google Scholar]
- Yokota J., Yamamoto T., Miyajima N., Toyoshima K., Nomura N., Sakamoto H., Yoshida T., Terada M., Sugimura T. Genetic alterations of the c-erbB-2 oncogene occur frequently in tubular adenocarcinoma of the stomach and are often accompanied by amplification of the v-erbA homologue. Oncogene. 1988 Mar;2(3):283–287. [PubMed] [Google Scholar]
- Yokota J., Yamamoto T., Toyoshima K., Terada M., Sugimura T., Battifora H., Cline M. J. Amplification of c-erbB-2 oncogene in human adenocarcinomas in vivo. Lancet. 1986 Apr 5;1(8484):765–767. doi: 10.1016/s0140-6736(86)91782-4. [DOI] [PubMed] [Google Scholar]
- Yonemura Y., Ninomiya I., Yamaguchi A., Fushida S., Kimura H., Ohoyama S., Miyazaki I., Endou Y., Tanaka M., Sasaki T. Evaluation of immunoreactivity for erbB-2 protein as a marker of poor short term prognosis in gastric cancer. Cancer Res. 1991 Feb 1;51(3):1034–1038. [PubMed] [Google Scholar]