Gastrin gene expression in human colon cancer cells measured by a simple competitive PCR method

Zhidong Xu, Bosong Dai, Bharati Dhruva, Pomila Singh

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Gastrin is mitogenic for several colon cancers and is postulated as an autocrine growth factor for colon cancer cells. In the present study we report the development of a simple competitive polymerase chain reaction (PCR) method for measuring relative abundance of gastrin gene expression in colon cancer cells. Primers flanking exons 2 and 3 of the gastrin gene were utilized for co-amplification of cDNA and genomic DNA. The amplification of genomic DNA was distinguished from that of cDNA by the presence of the 130 bp intron sequence which was resolved by electrophoresis on agarose gels. A standard reaction of competitive PCR, using known concentrations of genomic DNA and cDNA, was first established. The steady state levels of gastrin mRNA were next quantitated in three human colon cancer cell lines (HCT-116, Colo-205 and DLD-1) by competitive PCR. Gastrin mRNA levels in these cell lines ranged from ∼0.1 to 1.0 fmoles/mg total RNA (∼2-25 copies of gastrin mRNA per cell). This low to moderate levels of gastrin were expressed by human colon cancer cell lines which may function as autocrine growth factors for colon cancers.

Original languageEnglish (US)
Pages (from-to)671-678
Number of pages8
JournalLife Sciences
Volume54
Issue number10
DOIs
StatePublished - 1994

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Polymerase chain reaction
Gastrins
Gene expression
Colonic Neoplasms
Cells
Gene Expression
Polymerase Chain Reaction
Complementary DNA
Cell Line
Messenger RNA
Amplification
DNA
Intercellular Signaling Peptides and Proteins
Agar Gel Electrophoresis
Electrophoresis
Sepharose
Introns
Exons
Genes
Gels

ASJC Scopus subject areas

  • Pharmacology

Cite this

Gastrin gene expression in human colon cancer cells measured by a simple competitive PCR method. / Xu, Zhidong; Dai, Bosong; Dhruva, Bharati; Singh, Pomila.

In: Life Sciences, Vol. 54, No. 10, 1994, p. 671-678.

Research output: Contribution to journalArticle

Xu, Zhidong ; Dai, Bosong ; Dhruva, Bharati ; Singh, Pomila. / Gastrin gene expression in human colon cancer cells measured by a simple competitive PCR method. In: Life Sciences. 1994 ; Vol. 54, No. 10. pp. 671-678.
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