Method for quantitative measurement of gene expression for indentifying individuals at risk for bronchogenic carcinoma

Abstract

A method measure expression of multiple target genes in a progenitor cell for bronchogenic carcinoma comprising the use of reverse transcription-polymerase chain reaction (RT-PCR) to allow simultaneous expression measurement of the multiple target genes is disclosed.

Description

TECHNICAL BACKGROUND[0002] The PCR techniques are generally described in U.S. Pat. Nos. 4,683,195; 4,683,202 and 4,965,188. The PCR technique generally involves a process for amplifying any desired specific nucleic acid sequence contained within a nucleic acid molecule. The PCR process includes treating separate complementary strains of the nucleic acid with an excess of two oligonucleotide primers. The primers are extended to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence. The PCR process is carried out in a simultaneous step-wise fashion and can be repeated as often as desired in order to achieve increased levels of amplification of the desired nucleic acid sequence. According to the PCR process, the sequence of DNA between the primers on the respective DNA strains are amplified selectively over the remaining portions of the DNA and selected sample. The PCR process provides for the specific amplification of a ...

AI Technical Summary

Problems solved by technology

Normal bronchial epithelial cells (NBECs) are at an increased risk for oxidative damage following inhalational exposure to reactive oxygen species in cigarette smoke (1, 2), ozone (3), possibly asbestos (4), and other particulates in the environment.

These procarcinogens may be metabolically activated in the cytoplasm and subsequently damage nuclear DNA.

There is very little information presently available regarding quantitative levels of GST or glutathione peroxidase gene expression in primary NBECs relative to primary bronchogenic carcinoma tissue.

Consequently, studies that do not control for expression of all relevant genes may generate data that are difficult to interpret.

However, techniques described thus far allow only qualitative, not quantitative measurement.

Because GSHPx and GSTM3 each have peroxidase activity, cells expressing low levels of these genes are more susceptible to oxidant damage and carcinogenic transformation.

Further, GSTM3 and GSTP1 metabolically inactivate PAH diol-epoxide carcinogens in NBECs; thus, decreased expression levels in NBECs lead to a decrease in the cellular capacity to detoxify these carcinogens.

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