Establishment of a new method, transcription-reverse transcription concerted reaction, for detection of plasma hnRNP B1 mRNA, a biomarker of lung cancer.

Abstract

PURPOSE:

Development of an early detection marker is one of the most important strategies for improving overall prognosis in lung cancer patients. We previously reported that hnRNP B1--an RNA binding protein--is overexpressed in lung cancer tissue from the early stage of cancer, and found that hnRNP B1 mRNA is detectable in the plasma of lung cancer patients using real-time RT-PCR. The purpose of this study was to establish a quick and simple method for detecting plasma hnRNP B1mRNA for use in screening for lung cancer.

METHODS:

TRC, a homogenous method for fluorescence real-time monitoring of isothermal RNA amplification using intercalation activating fluorescence DNA probe, was used to detect plasma hnRNP B1 mRNA.

RESULTS:

The detection limit of hnRNP B1 mRNA by TRC using synthetic control RNA or total RNA derived from a lung cancer cell line was 25 or 8.65 x 10(2) copies, respectively. Using total RNA extracted from 600 mul of plasma, we detected hnRNP B1 mRNA in 39.1% (9/23) of lung cancer patients, with levels ranging from 1.9 to 19,045.5 copies/100 ng RNA, and in 5.2% (5/97) of healthy volunteers. Copy numbers were not associated with age, gender, smoking status, or histological type of cancer. TRC could detect 10(3) copies of hnRNP B1 mRNA in 10 min.

CONCLUSION:

Detection of plasma hnRNP B1 mRNA by TRC is a quick, easy, and non-invasive method suitable for lung cancer screening.

• Authors: Akemi Sato, Naoko Sueoka-Aragane, Juichi Saitoh, Kazutoshi Komiya, Takashi Hisatomi, Rika Tomimasu, Shinichiro Hayashi, Eisaburo Sueoka
• Journal: Journal of cancer research and clinical oncology (J Cancer Res Clin Oncol) Vol. 134 Issue 11 Pg. 1191-7 (Nov 2008) ISSN: 1432-1335 [Electronic] Germany
• PMID: 18461365 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA Primers
• Genetic Markers
• Heterogeneous-Nuclear Ribonucleoprotein Group A-B
• Heterogeneous-Nuclear Ribonucleoproteins
• RNA, Messenger
• RNA, Neoplasm
• hnRNP A2
• RNA

Topics

• Base Sequence
• DNA Primers
• Female
• Genetic Markers
• Heterogeneous-Nuclear Ribonucleoprotein Group A-B (blood, genetics)
• Heterogeneous-Nuclear Ribonucleoproteins (genetics)
• Humans
• Lung Neoplasms (genetics)
• Male
• Molecular Sequence Data
• Prognosis
• RNA (genetics)
• RNA, Messenger (genetics)
• RNA, Neoplasm (genetics)
• Reference Values
• Reverse Transcriptase Polymerase Chain Reaction
• Sensitivity and Specificity