Advertisement

In vitro expression of the diphtheria toxin A-chain gene under the control of human chorionic gonadotropin gene promoters as a means of directing toxicity to ovarian cancer cell lines

      Abstract

      Objective: Our goal was to determine whether toxicity of the diphtheria toxin A-chain gene regulated by the human chorionic gonadotropin promoter can be directed to malignant ovarian cell lines. Study design: Plasmids containing diphtheria toxin A-chain gene linked to the regulatory elements of the metalloergothioneine and human chorionic gonadotropin promoters were transfected into the cell lines. Expression of diphtheria toxin A-chain gene was determined by the inhibition of a cotransfected luciferase reporter gene. Results: Cytotoxicity of the diphtheria toxin A-chain gene is shown in a dose-responsive manner. Transfection of a plasmid expressing the diphtheria toxin A-chain gene controlled by a constitutive promoter readily inhibits protein synthesis. Specific inhibition of luciferase protein synthesis occurs in ovarian cancer cells transfected with the diphtheria toxin A-chain gene under the control of the human chorionic gonadotropin promoters when compared with normal ovarian epithelial cells or fibroblasts. Conclusions: These data demonstrate the preferential expression of the diphtheria toxin A-chain gene, regulated by the human chorionic gonadotropin promoter, to ovarian cancer cell lines. This provides an avenue for targeting such cells for suicide, toxin, or cytokine genes.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to American Journal of Obstetrics & Gynecology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Parker SL
        • Tong T
        • Bolden S
        • Wingo PA
        Cancer Statistics, 1996.
        CA Cancer J Clin. 1996; 46: 5-27
        • Cannistra SA
        Cancer of the ovary.
        N Engl J Med. 1993; 329: 1550-1559
        • Lidor Y
        • Bast Jr, RC
        Serotherapy of ovarian cancer.
        Natl Immun Cell Growth Regul. 1988; 7: 193-215
        • Breitman ML
        • Clapoff S
        • Rossant J
        • Tsui LC
        • Glode LM
        • Maxwell IH
        • et al.
        Genetic ablation: targeted expression of a toxin gene causes microphthalmia in transgenic mice.
        Science. 1987; 238: 1563-1565
        • Cole LA
        • Wang YX
        • Elliott M
        • Latif M
        • Chambers JT
        • Chambers SK
        • et al.
        Urinary human chorionic gonadotropin free beta-subunit and beta-core fragment: a new marker of gynecological cancers.
        Cancer Res. 1988; 48: 1356-1360
        • Marcillac I
        • Troalen F
        • Bidart JM
        • Ghillani P
        • Ribrug V
        • Escudier B
        • et al.
        Free human chorionic gonadotropin beta subunit in gonadal and nongonadal neoplasms.
        Cancer Res. 1992; 52: 3901-3907
        • Cosgrove DE
        • Campain JA
        • Cox GS
        Chorionic gonadotropin synthesis by human tumor cell lines: examination of subunit accumulation, steady-state levels of mRNA, and gene structure.
        Blochim Biophys Acta. 1989; 1007: 44-54
        • Wehmann RE
        • Nisula BC
        Characterization of a discrete degradation product of the human chorionic gonadotropin beta-subunit in humans.
        J Clin Endocrinol Metab. 1980; 51: 101-105
        • Behrens BC
        • Hamilton TC
        • Masuda H
        • Grotzinger KR
        • Whang-Peng J
        • Louie KG
        • et al.
        Characterization of a cis-diamminedichloroplatinum(ll)-resistant human ovarian cancer cell line and its use in evaluation of platinum analogues.
        Cancer Res. 1987; 47: 414-418
        • Bast Jr, RC
        • Feeney M
        • Lazarus H
        • Nadler LM
        • Colvin RB
        • Knapp RC
        Reactivity of a monoclonal antibody with human ovarian carcinoma.
        J Clin Invest. 1981; 68: 1331-1337
        • Lidor YJ
        • Xu FJ
        • Martinez-Maza O
        • Olt GJ
        • Marks JR
        • Berchuck A
        • et al.
        Constitutive production of macrophage colony-stimulating factor and interleukin-6 by human ovarian surface epithelial cells.
        Exp Cell Res. 1993; 207: 332-339
        • de Wet JR
        • Wood KV
        • DeLuca M
        • Helinski DR
        • Subramani S
        Firefly luciferase gene: structure and expression in mammalian cells.
        Mol Cell Biol. 1987; 7: 725-737
        • Maxwell IH
        • Maxwell F
        • Glode LM
        Regulated expression of a diphtheria toxin A-chain gene transfected into human cells: possible strategy for inducing cancer cell suicide.
        Cancer Res. 1986; 46: 4660-4664
        • Fiddes JC
        • Talmadge K
        Structure, expression, and evolution of the genes for the human glycoprotein hormones.
        Recent Prog Horm Res. 1984; 40: 43-78
        • Maxwell IH
        • Harrison GS
        • Wood WM
        • Maxwell F
        A DNA cassette containing a trimerized SV40 polyadenylation signal which efficiently blocks spurious plasmid initiated transcription in transfected mammalian cells.
        Biotechniques. 1989; 7: 276-280
        • Bast Jr, RC
        • De Fabritiis P
        • Lipton J
        • Gelber R
        • Maven C
        • Nadler L
        • et al.
        Elimination of malignant clonogenic cells from human bone marrow using multiple monoclonal antibodies and complement.
        Cancer Res. 1985; 45: 499-503
        • Lidor YJ
        • Shpall EJ
        • Peters WP
        • Bast Jr, RC
        Synergistic cytotoxicity of different alkylating agents for epithelial ovarian cancer.
        Int J Cancer. 1991; 49: 704-710
        • Maxwell IH
        • Maxwell F
        Electroporation of mammalian cells with a firefly luciferase expression plasmid: kinetics of transient expression differ markedly among cell types.
        DNA. 1988; 7: 557-562
        • McGregor WG
        • Kuhn RW
        • Jaffe RB
        Biologically active chorionic gonadotropin: synthesis by the human fetus.
        Science. 1983; 220: 306-308
        • Iles RK
        • Purkis PE
        • Whitehead PC
        • Oliver RT
        • Leigh I
        • Chard T
        Expression of beta human chorionic gonadotrophin by non-trophoblastic non-endocrine “normal” and malignant epithelial cells.
        Br J Cancer. 1990; 61: 663-666
        • Goldstein S
        • Jones RA
        • Hardin JW
        • Braunstein GD
        • Shmookler Reis RJ
        Expression of alpha- and beta-human chorionic gonadotropin subunits in cultured human cells.
        In Vitro Cell Dev Biol. 1990; 26: 857-864
        • Kendall SK
        • Saunders TL
        • Jin L
        • Lloyd RV
        • Glode LM
        • Nett TM
        • et al.
        Targeted ablation of pituitary gonadotropes in transgenic mice.
        Mol Endocrinol. 1991; 5: 2025-2036
        • Nilson JH
        • Bokar JA
        • Andersen B
        • Bohinski R
        • Kennedy G
        • Kevi RA
        • et al.
        CRE-binding proteins interact cooperatively to enhance placental-specific expression of the glycoprotein hormone alpha-subunit gene.
        Ann N Y Acad Sci. 1989; 564: 77-85
        • Hamemik DL
        • Keri RA
        • Clay CM
        • et al.
        Gonadotrope- and thyrotrope-specific expression of the human and bovine glycoprotein hormone alpha-subunit genes is regulated by distinct cis-acting elements.
        Mol Endocrinol. 1992; 6: 1745-1755
        • Sarapura VD
        • Wood WM
        • Gordon DF
        • Ocran KW
        • Kao MY
        • Ridgway EC
        Thyrotrope expression and thyroid hormone inhibition map to different regions of the mouse glycopretein hormone alpha-subunit gene promoter.
        Endocrinology. 1990; 127: 1352-1361