MspI restriction fragment length polymorphism at the glycoprotein hormone α-subunit locus. Association of certain genotypes with neoplasia

G. Stanley Cox, Dominic E Cosgrove, Michael J. Haas, Warren Stiles, David G. McIntosh

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

A restriction fragment length polymorphism (RFLP) in the human glycoprotein hormone common α-subunit gene has been identified and partially characterized in normal lymphocytes and placentae, established tumor cell lines, and tumor biopsy samples. High molecular weight DNA was digested with the restriction endonuclease MspI, separated by electrophoresis in agarose gels, transferred to nylon membranes by the method of Southern, and hybridized to 32P-labeled human chorionic gonadotropin α-subunit cDNA. After autoradiography, bands were detected at 5.3, 3.3, 2.1, 1.6, 0.8 and 0.6 kbp. Presence of the 5.3, 3.3 and 0.6 kbp bands was invariant and uninformative. Patterns missing the 0.8 kbp band and both the 2.1 and 1.6 kbp bands are consistent with separate alleles that occur in placental and lymphocyte DNA with frequencies of 0.44 (15/34) and 0.06 (2/34), respectively. Presence of all three bands (2.1, 1.6 and 0.8 kbp) is indicative of heterozygosity, occurring at a frequency of 0.50 (17/34). Additional restriction patterns, not yet observed in DNA isolated from term placentae or circulating lymphocytes, were detected in DNA obtained from tumor cell lines and fresh tumor tissues at frequencies of 0.79 (15/19) and 0.59 (10/17), respectively. Thus, particular α-subunit genotypes are disproportionately represented in tumor-derived DNA, occurring at frequencies 10- to 13-times higher than would be predicted from their occurrence in normal tissue. Paired normal and tumor tissues from the same individual exhibited identical hybridization patterns, suggesting that this RFLP may be representative of a predisposition toward a variety of neoplasias rather than indicative of a change in DNA structure at or near this locus as a result of tumor development.

Original languageEnglish (US)
Pages (from-to)83-96
Number of pages14
JournalBiochimica et Biophysica Acta - Gene Structure and Expression
Volume1354
Issue number1
DOIs
StatePublished - Oct 1 1997

Fingerprint

Polymorphism
Restriction Fragment Length Polymorphisms
Tumors
Glycoproteins
Genotype
Hormones
DNA
Lymphocytes
Neoplasms
Tumor Cell Line
Placenta
Tissue
Cells
Agar Gel Electrophoresis
Nylons
DNA Restriction Enzymes
Chorionic Gonadotropin
Autoradiography
Biopsy
Electrophoresis

Keywords

  • Chromosome 6
  • RFLP
  • α-Choriogonadotropin

ASJC Scopus subject areas

  • Structural Biology
  • Biophysics
  • Biochemistry
  • Genetics

Cite this

MspI restriction fragment length polymorphism at the glycoprotein hormone α-subunit locus. Association of certain genotypes with neoplasia. / Cox, G. Stanley; Cosgrove, Dominic E; Haas, Michael J.; Stiles, Warren; McIntosh, David G.

In: Biochimica et Biophysica Acta - Gene Structure and Expression, Vol. 1354, No. 1, 01.10.1997, p. 83-96.

Research output: Contribution to journalArticle

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abstract = "A restriction fragment length polymorphism (RFLP) in the human glycoprotein hormone common α-subunit gene has been identified and partially characterized in normal lymphocytes and placentae, established tumor cell lines, and tumor biopsy samples. High molecular weight DNA was digested with the restriction endonuclease MspI, separated by electrophoresis in agarose gels, transferred to nylon membranes by the method of Southern, and hybridized to 32P-labeled human chorionic gonadotropin α-subunit cDNA. After autoradiography, bands were detected at 5.3, 3.3, 2.1, 1.6, 0.8 and 0.6 kbp. Presence of the 5.3, 3.3 and 0.6 kbp bands was invariant and uninformative. Patterns missing the 0.8 kbp band and both the 2.1 and 1.6 kbp bands are consistent with separate alleles that occur in placental and lymphocyte DNA with frequencies of 0.44 (15/34) and 0.06 (2/34), respectively. Presence of all three bands (2.1, 1.6 and 0.8 kbp) is indicative of heterozygosity, occurring at a frequency of 0.50 (17/34). Additional restriction patterns, not yet observed in DNA isolated from term placentae or circulating lymphocytes, were detected in DNA obtained from tumor cell lines and fresh tumor tissues at frequencies of 0.79 (15/19) and 0.59 (10/17), respectively. Thus, particular α-subunit genotypes are disproportionately represented in tumor-derived DNA, occurring at frequencies 10- to 13-times higher than would be predicted from their occurrence in normal tissue. Paired normal and tumor tissues from the same individual exhibited identical hybridization patterns, suggesting that this RFLP may be representative of a predisposition toward a variety of neoplasias rather than indicative of a change in DNA structure at or near this locus as a result of tumor development.",
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AU - Cox, G. Stanley

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AB - A restriction fragment length polymorphism (RFLP) in the human glycoprotein hormone common α-subunit gene has been identified and partially characterized in normal lymphocytes and placentae, established tumor cell lines, and tumor biopsy samples. High molecular weight DNA was digested with the restriction endonuclease MspI, separated by electrophoresis in agarose gels, transferred to nylon membranes by the method of Southern, and hybridized to 32P-labeled human chorionic gonadotropin α-subunit cDNA. After autoradiography, bands were detected at 5.3, 3.3, 2.1, 1.6, 0.8 and 0.6 kbp. Presence of the 5.3, 3.3 and 0.6 kbp bands was invariant and uninformative. Patterns missing the 0.8 kbp band and both the 2.1 and 1.6 kbp bands are consistent with separate alleles that occur in placental and lymphocyte DNA with frequencies of 0.44 (15/34) and 0.06 (2/34), respectively. Presence of all three bands (2.1, 1.6 and 0.8 kbp) is indicative of heterozygosity, occurring at a frequency of 0.50 (17/34). Additional restriction patterns, not yet observed in DNA isolated from term placentae or circulating lymphocytes, were detected in DNA obtained from tumor cell lines and fresh tumor tissues at frequencies of 0.79 (15/19) and 0.59 (10/17), respectively. Thus, particular α-subunit genotypes are disproportionately represented in tumor-derived DNA, occurring at frequencies 10- to 13-times higher than would be predicted from their occurrence in normal tissue. Paired normal and tumor tissues from the same individual exhibited identical hybridization patterns, suggesting that this RFLP may be representative of a predisposition toward a variety of neoplasias rather than indicative of a change in DNA structure at or near this locus as a result of tumor development.

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