Faculty Collaboration Database

Detection of oncogene mRNA sequences in cultured cells by in situ hybridization. Ann Clin Lab Sci 1987;17(2):74-82

Date

03/01/1987

Pubmed ID

3579212

Scopus ID

2-s2.0-0023127121 (requires institutional sign-in at Scopus site)   1 Citation

Abstract

The present study was undertaken to determine the feasibility of using in situ hybridization techniques to identify oncogene transcription in cultured cells. Following in situ hybridization with 32P-labeled v-src and v-Ha-ras DNA probes, src and Ha-ras related transcripts were identified in cell lines transfected with v-src and Ha-ras, respectively. In both the v-src and c-Ha-ras transfected cell lines, the number of silver grains over individual cells were significantly higher (p less than 0.001, t-test) than in a non-transfected, non-tumorigenic, rat esophageal epithelial cell line. There was a highly variable number of silver grains above individual cells. Significantly fewer silver grains were counted over cells that had been preincubated with either non-labeled v-src or v-Ha-ras DNA or that were pretreated with RNase A. Both oncogene transfected cell lines contained approximately 10 times more oncogene related mRNAs than non-transfected cells as judged by the numbers of silver grains over individual cells. Filter-hybridization analysis of the transfected and non-transfected cell lines confirmed that the expression of src and Ha-ras transcripts was higher in the transfected cell lines than in the non-transfected cell line. Therefore, the in situ hybridization technique would appear useful for the identification of oncogene transcripts in single cells and could potentially be applied to cytological preparations of human cells and to human tumor cells in culture.

Author List

Stoner GD, You M, Skouv J, Budd GC, Pansky B, Wang Y

MESH terms used to index this publication - Major topics in bold

Animals
Autoradiography
Base Sequence
Cell Line
Cell Transformation, Neoplastic
Cell Transformation, Viral
DNA
Humans
Mice
Nucleic Acid Hybridization
Oncogenes
RNA, Messenger
Rats
Transcription, Genetic
Transfection