Identification of mRNAs differentially-expressed between benign and malignant breast tumour cells

Abstract

Two suppression subtracted cDNA libraries have been constructed, one containing cDNAs to mRNAs present at a higher level in a benign human breast tumour-derived cell line relative to the malignant mammary cell line, MCF-7, and the other containing cDNAs present at a higher level in the MCF-7 cells relative to the benign cells. Randomly-picked cloned DNAs have been sequenced yielding 29 and 128 different cDNAs from the benign and malignant libraries, respectively. Using reverse Northern hybridisation, 76% and 83% of the cDNAs were differentially expressed by greater than two-fold, whilst 14% and 11% of cDNAs in the respective libraries were differentially expressed by more than 15-fold. Amongst these were oestrogenresponsive cDNAs and expressed sequence tags. One such oestrogen-responsive expressed sequence tag, M41, is transcribed from a gene located on chromosome 21q22.3, within an intron of a larger gene. The M41 gene contains oestrogen response elements, one of which is associated with alu repeats. M41 mRNA is expressed at a statistically significantly higher level in human breast cancer specimens than in normal human breast and benign lesions. In carcinomas, its up-regulation is associated with the development of the malignant cell.

Using a Xenograft Model of Human Breast Cancer Metastasis to Find Genes Associated with Clinically Aggressive Disease

“…In this report, the gene expression patterns in isogenic variants of a human breast cancer cell line with different metastatic potential were compared, and 106 genes (0.66% of the total analyzed) were identified as differentially expressed >2.5-fold. The relatively small number is likely due to the common origin of the cell lines, decreasing the genetic variation that may be found in comparisons of cell lines from different individuals (32). However, the fact that the expression levels of 106 genes were altered z2.5-fold in cells with increased metastatic potential indicates that multiple genetic events contribute to metastasis.…”

DSCAM‐AS1 regulates the G1/S cell cycle transition and is an independent prognostic factor of poor survival in luminal breast cancer patients treated with endocrine therapy

“…DSCAM‐AS1 was originally described by Liu. It is transcribed from a gene located on chromosome 21q22.3 (GRCh38/hg38) with a full length of approximately 1.4 kb . In 2015, Miano et al analyzed the RNA‐seq data from MCF‐7 cells in the absence of estrogen.…”

Human Homologue of Cement Gland Protein, a Novel Metastasis Inducer Associated with Breast Carcinomas

“…A suppression subtractive (11) library consisting of PCR products representing mRNAs expressed at a higher level in the malignant breast epithelial cell line, MCF-7, relative to a benign human breast-derived cell line, Huma 123, was constructed using a PCR-Select cDNA Subtraction kit (Clontech, Palo Alto, CA) as described previously (14). Reverse Northern screening of the subtracted cDNA library was carried out as described previously (12).…”

“…PCR-selected suppression subtractive hybridization (11) has been used to identify cDNAs representing mRNAs differentially expressed (12)(13)(14) between an estrogen receptor a (ERa)-negative benign human mammary epithelial cell line, Huma 123 (15), and the ERa-positive malignant human mammary epithelial cell line, MCF-7 (16). The resulting subtracted libraries contained wellcharacterized, differentially expressed cDNAs that have been associated previously with tumor progression (12).…”