비소세포 폐암환자에서 Real-time PCR을 이용한 혈장 Heterogeneous Ribonucleoprotein B1 mRNA 상대정량의 유용성
Comparative Quantification of Plasma hnRNP B1 mRNA in Non-small Cell Lung Cancer Patients by Real-time PCR
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배경 : 혈장에서 검출되는 핵산(cell-free nucleic acids)이 최근 암의 진단에 있어 비침습적 표지자로 유용한 것으로 알려져 있다. Nuclear core complex의 하나인, heterogeneous nuclear ribonucleoprotein (hnRNP) B1은 폐암 초기에 과발 현 되는 것으로 알려져 있다. 본 연구에서는 한국에서 유병률이 높고, 종종 폐암과 구별이 어려운 폐결핵을 포함한 양성폐질환 으로부터 비소세포 폐암환자를 확인하는데 혈장 hnRNP B1 mRNA가 유용한지 알아보고자 하였다 방법 : 비소세포 폐암환자 30명, 폐결핵을 포함한 양성 폐질 환자 30명, 정상인 10명을 대상으로 혈장에서 RNA를 추출하였 다. 혈장hnRNP B1 mRNA를 측정하기 위해 predesigned, validated TaqMan Gene Expression Assay (Applied Biosystems, USA)를 이용하였고, 상대적인 정량을 위해 pre-developed beta-actin (ACTB) mRNA을 사용하였다. Delta-delta Ct 방법을 이용하여 상대적으로 정량하였다. 결과 : 비소세포 폐암환자 93.3% (28/30)에서 혈장 hnRNP B1 mRNA가 측정되었다. 정상인의 hnRNP B1 mRNA로 보정 한 상대 발현 값인 2-ΔΔCt는 비소세포 폐암환자군에서 62.2 (95% 신뢰구간; 6.4-210.1), 양성폐질환자군에서 2.7 (95% 신뢰구간; 0.5-13.6)로 통계적으로 유의하게 높게 발현되었다(P<0.001, Mann-Whitney U test). 결론 : 혈장 hnRNP B1 mRNA는 정상대조군 또는 양성폐질 환군에 비해 비소세포 폐암환자에서 유의하게 증가되어 있었다. 따라서, 혈장 hnRNP B1 mRNA는 암표지자로서 비소세포성 폐암의 검출에 유용할 수 있을 것으로 판단된다.
Abstract
Background : Circulating cell-free nucleic acids are known to be a noninvasive diagnostic tool for cancer detection. Heterogeneous nuclear ribonucleoprotein (hnRNP) B1, a nuclear core complex, is overexpressed in early stage lung cancer. We intended to evaluate the usefulness of plasma hn-RNP B1 mRNA in differentiating non-small cell lung cancer (NSCLC) from other benign lung diseases, especially pulmonary tuberculosis, which is highly prevalent in Korea and often difficult to distinguish from lung cancer. Methods : Plasma RNA was extracted from 30 patients with NSCLC, 30 patients with benign lung diseases including pulmonary tuberculosis, and 10 healthy controls. Plasma hnRNP B1 mRNA was measured by TaqMan Gene Expression Assay (Applied Biosystems, USA), and pre-developed betaactin (ACTB) mRNA was used for normalization. We analyzed the relative gene expression data using the delta-delta Ct method. Results : Plasma hnRPN B1 mRNA was measurable in 93.3% (28/30) of NSCLC patients. Normalized 2-ΔΔCt of plasma hnRPN B1 mRNA was 62.2 (95%Cl, 6.4-210.1) in NSCLC patients and 2.7 (95% Cl, 0.5-13.6) in benign lung disease patients (P<0.001, Mann-Whitney U test). Conclusions : Plasma hnRNP B1 mRNA was significantly increased in patients with lung cancer compared with that in patients with other benign lung diseases. Plasma hnRNP B1 mRNA may be useful as a potential marker for the detection of NSCLC.
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