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Capture‐RT‐PCR assay of mRNA in breast tumors: int ‐2 and HERV‐K env mRNA
Author(s) -
Gillespie David,
Poirer Stephanie,
Poley Justin,
Cuddy Kevin,
Brodsky Isadore,
Brodsky Jeffrey
Publication year - 1993
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240531152
Subject(s) - gene expression , breast cancer , biology , biomarker , real time polymerase chain reaction , cancer research , cancer , messenger rna , gene , malignancy , microbiology and biotechnology , genetics
Abstract Gene expression at the mRNA level is likely to be a rapidly responding intermediate biomarker for use as a surrogate endpoint in Phase II clinical trials of breast cancer. Using a technology called “capture‐RT‐PCR,” we have been able to show ectopic int ‐2 mRNA expression in 7/9 breast tumors, including two fibroadenomas. The level of expression varied, high expression being observed in three tumors. Frequent expression of int ‐2 in breast tumors is contrary to published reports, possibly reflecting differences in technology. int ‐2 expression in fibroadenomas suggests that the marker may precede malignancy. Conversely, if fibroadenomas are not premalignant, int ‐2 expression may be gratuitous. These alternatives need to be tested in a clinical trial. Human endogenous retrovirus K (HERV‐K) env gene expression was detected in only 1/7 breast tumors. Expression of this retroviral gene is more restricted than int ‐2 expression. c‐ myc was expressed in all 10 tumors studied, albeit at widely varying levels. Expression of prad 1 and erb B‐2 are under study. Gene expression will be compared with gene amplification. It is anticipated that a capture‐RT‐PCR assay simultaneously signifying levels of expression of several oncogenes/anti‐oncogenes will be a most informative surrogate endpoint biomarker. In particular, chaotropic salt methods that simplify sample preparation and mRNA isolation and reduce these combined steps to a 10 minute procedure will be presented.