Establishment Of Quantification In FQ-RT-PCR To Measure The HnRNP A2/B1 In The Peripheral Blood And Clinical Value Between The Level Of HnRNPA2/B1 Expression And The Diagnosis Of Lung Cancer

Heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2/B1) is a kind of cancer embryo antigen relating to the beginning and the development of lung cancer. It has been recently reported that hnRNPA2/Bl pooly expressed in normal cells and overexpressed in the beginning of lung cancer . the level of expression of hnRNPA2/Bl rised in the developing process of lung cancer.After reaching a peak value it reduced on a poor level.IHC(Imrnunohistochemistry) and classical RT-PCR(Reverse transcription polymerase chain reaction) are often adopted in detection of nature determination or quasi-quantification of hnRNP A2/B1 based on the overexpression of protein and alteration of mRNA respectively. We investigate the difference of expression among patients of lung cancer and other two groups by fluorescent quantification Real-time polymerase chain reaction(FQ-RT-PCR) to find the clinical value in diagnosis of lung cancer.1 Establishment of FQ-RT-PCR for quantitatively measuring the level of hnRNP A2 /Bl in the peripheral bloodRecently, besides the detection of circulating tumor cells in peripheral blood of patients with solid tumors, the presence of free circulating nucleic acids in the plasma and serum has also been reported. This project is focused on the study of isolating and amplifying intact extracellular tumor-related mRNA from the plasma of patients with lung cancer and on the establishment of FQ-RT-PCR based amplification systems. A TaqMan probe technique based on ABI 5700 Fluorescent Quantification PCR System was introduced to quantify the expression of hnRNP A2/B1 in peripheral blood. The method is: series dilutions of a plasmid with known concentration were used to establish the standard curve and to measure the amplification efficiency of the system. The standard curve was defined by six points. The amplification efficiency was close to 100% by modifying reaction...