M. Sternlicht et al., A NOVEL STRATEGY FOR THE INVESTIGATION OF CLONALITY IN PRECANCEROUS DISEASE STATES AND EARLY STAGES OF TUMOR PROGRESSION, Biochemical and biophysical research communications, 199(2), 1994, pp. 511-518
A novel strategy for clonality determination from only 100 cells using
the polymerase chain reaction in amplifying a 511 bp region located w
ithin the first intron of the human hypoxanthine phosphoribosyl transf
erase (HPRT) gene has been devised. The strategy rests on several obse
rvations: that this region in females contains two HpaII/MspI sites wh
ose methylation remains both obligate with X chromosome inactivation a
nd independent of tumor progression; and that this region contains sin
gle base allelic polymorphisms in 5-10% of females which can be detect
ed by denaturing gradient gel electrophoresis (DGGE) on the PCR produc
t. In polymorphic individuals, multiple bands (homo- and heteroduplexe
s) indicate multiclonality, single bands indicate monoclonality, and t
heir comparative migrations indicate clonal identity/non-identity. (C)
1994 Academic Press, Inc.