A novel strategy for the investigation of clonality in precancerous disease states and early stages of tumor progression.
A novel strategy for clonality determination from only 100 cells using the polymerase chain reaction in amplifying a 511 bp region located within the first intron of the human hypoxanthine phosphoribosyl transferase (HPRT) gene has been devised. The strategy rests on several observations: that this region in females contains two HpaII/MspI sites whose methylation remains both obligate with X chromosome inactivation and independent of tumor progression; and that this region contains single base allelic polymorphisms in 5-10% of females which can be detected by denaturing gradient gel electrophoresis (DGGE) on the PCR product. In polymorphic individuals, multiple bands (homo- and heteroduplexes) indicate multiclonality, single bands indicate monoclonality, and their comparative migrations indicate clonal identity/non-identity.
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Department of Pathology, UCLA School of Medicine 90024.
Mirell
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Safarians
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Barsky
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D001483:Base Sequence; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D002999:Clone Cells; D017931:DNA Primers; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific; D005260:Female; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D007438:Introns; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology*; D016376:Oligonucleotides, Antisense; D010735:Phosphoglycerate Kinase / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011230:Precancerous Conditions / Q000201:enzymology / Q000235:genetics* / Q000473:pathology*; D014960:X Chromosome
D017931:DNA Primers; D016376:Oligonucleotides, Antisense; D007041:Hypoxanthine Phosphoribosyltransferase; D010735:Phosphoglycerate Kinase; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific