Abstract
Protein extracted from conventional formalin-fixed and paraffin-embedded tissue sections of human gliomas was examined for immunoblot analysis using antibody against the placental form of glutathioneS-transferase (GST-π). Four benign astrocytomas, five anaplastic astrocytomas and four glioblastomas were used in this study. The preliminary study demonstrated that immunoreactivity of GST-π was well preserved in normal brain tissue and normal term placenta fixed in acetone, formalin or buffered formalin (pH 7.4). GST-π in gliomas fixed in formalin also had a good immunoreactivity and showed clear bands on nitrocellulose membranes processed by the method of Western blotting using anti-GST-π antibody. The results of immunoblot analysis for GST-π indicate that the intensity of immunoreactivity of benign astrocytoma, anaplastic astrocytoma and glioblastoma increases with the advance of malignancy of these neoplasms. Western blot analysis for GST-π can be performed using protein extracted from formalin-fixed and paraffin-embedded tissue sections, and the immunoreactive bands can be analyzed quantitatively by densitometric scanning.
Similar content being viewed by others
Abbreviations
- GST-π:
-
placental form of glutathioneS-transferase
- SDS-PAGE:
-
sodium dodecylsulfate/polyacrylamide gel electrophoresis
References
Batist G, Tulpule A, Sinha BK, Katki AG, Myers CE, Cowan KH (1986) Overexpression of a novel anionic glutathione transferase in multidrug-resistant human breast cancer cells. J Biol Chem 261:15544–15549
Chasseaud LF (1979) The role of glutathione and glutathione S-transferases in the metabolism of chemical carcinogens and other electrophilic agents. Adv Cancer Res 29:175–274
Clark RK, Damjanov I (1986) Immunoblotting of keratin polypeptides extracted from tissue preserved in standard histologic fixatives. J Histochem Cytochem 34:679–682
Conti CJ, Larcher F, Chesner J, Aldaz CM (1988) Polyacrylamide gel electrophoresis and immunoblotting of proteins extracted from paraffin-embedded tissue sections. J Histochem Cytochem 36:547–550
Hara A, Sakai N, Yamada H, Tanaka T, Kato K, Mori H, Sato K (1989) Induction of glutathione S-transferase, placental type in T9 glioma cells by dibutyryladenosine 3′, 5′-cyclic monophosphate and modification of its expression by naturally occurring isothiocyanates. Acta Neuropathol 79:144–148
Hara A, Yamada H, Sakai N, Hirayama H, Tanaka T, Mori H (1990) Immunohistochemical demonstration of the placental form of glutathione S-transferase a detoxifying enzyme in human gliomas. Cancer 66:2563–2568
Jakoby WB (1978) The glutathione S-transferases: a group of multifunctional detoxification proteins. Adv Enzymol 46:383–414
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophoage T4. Nature 227:680–685
Mannervik B (1985) The isoenzymes of glutathione transferase. Adv Enzymol Relat Areas Mol Biol 57:357–417
McGown AT, Fox BW (1986) A proposed mechanism of resistance to cyclophosphamide and phosphoramide mustard in a Yoshida cell line in vitro. Cancer Chemother Pharmacol 17:223–226
Moorghen M, Cairns J, Forrester LM, Hayes JD, Hall, A, Cattan AR, Wolf CR, Harris AL (1991) Enhanced expression of glutathione S-transferase in colorectal carcinoma compared to non-neoplastic mucosa. Carcinogenesis 12:13–17
Moscow JA, Fairchild CR, Madden MJ, Ransom DT, Wieand HS, O'Brien EE, Poplack DG, Cossman J, Myers CE, Cowan KH (1989) Expression of anionic glutathione S-transferase and P-glycoprotein genes in human tissues and tumors. Can Res 49:1422–1428
Sato K, Kitahara A, Satoh K, Ishikawa, T, Tatematsu M, Ito N (1984) The placental form of glutathione S-transferase as a new marker protein for preneoplasia in rat chemical hepatocarcinogenesis. Gann 75:199–202
Satoh K, Kitahara A, Soma Y, Inaba Y, Hatayama I, Sato K (1985) Purification, induction, and distribution of placental glutathione transferase: a new marker enzyme for preneoplastic cells in the rat chemical hepatocarcinogenesis. Proc Natl Acad Sci USA 82:3964–3968
Schisselbauer JC, Silver R, Papadopoulos E, Abram K, LaCreta FP, Tew KD (1990) Characterization of glutathione S-transferase expression in lymphocytes from chronic lymphocytic leukemia patients. Cancer Res 50:3562–3568
Shea TC, Kelley SL, Henner WD (1988) Identification of an anionic form of glutathione transferase present in may human tumors and human tumor cell lines. Can Res 48:527–533
Tatematsu M, Mera Y, Ito N, Satoh K (1985) Relative merits of immunohistochemical demonstration of placental, A, B and C forms of glutathione S-transferase and histochemical demonstration of γ-glutamyl transferase as markers of altered foci during liver carcinogenesis in rats. Carcinogenesis 6:1621–1626
Tatematsu M, Aoki T, Kagawa M, Mera Y, Ito N (1988) Reciprocal relationship between developmental glutathione S-transferase positive liver foci and proliferation of surrounding hepatocytes in rats. Carcinogenesis 9:221–225
Tylor CW, Brattain MG, Yeoman LC (1985) Occurrence of cytosolic protein and phosphoprotein changes in human colon tumor cells with the development of resistance to Mitomycin C. Cancer Res 45:4422–4427
Towbin H, Staehelin T, Gordon J (1982) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350
Tsuchida S, Izumi T, Shimizu T, Ishikawa T, Hatayama I, Satoh K, Sato K (1987) Purification of a new acidic glutathione S-transferase, GST-Yn1 Yn1, with a high leukotriene-C4 synthase activity from rat brain. Eur J Biochem 170:159–164
Waxman DJ (1990) Glutathione S-transferase: role in alkylating agent resistance and possible target for modulation chemotherapy-review. Cancer Res 50:6449–6454
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hara, A., Sakai, N., Yamada, H. et al. Immunoblot analysis of the placental form of glutathione S-transferase in protein extracted from paraffin-embedded human glioma tissue. J Cancer Res Clin Oncol 119, 493–496 (1993). https://doi.org/10.1007/BF01215931
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01215931