Nine miRNAs expression differences and its significance in serum and peripheral blood mononuclear cell of lung cancer / 中华检验医学杂志

ABSTRACT

Objective To investigate the change of expression patterns of miRNA in the serum and peripheral blood mononuclear cells (PBMC) of lung cancer and its significance.Methods Clinical case control study was employed.Establish the method of microRNA(miRNA) detection by real time quantitative PCR (RT-qPCR).peripheral blood of the study subjects were collected in First affiliated hospital of SooChow University from November 2011 to September 2012.Gender and age matched subjects whose median age was 64(40-85) included 61 lung cancer cases,48 healthy control and benign lung diseases.We used quantitative RT-PCR to assess miRNA expression pattern of-miR-20a,21,-25,-29,-31,-126,-129,-145 and -205 in peripheral blood.U6 was taken as reference,and the expression of miRNA were indicated as F =2-△Ct,ACt =CtmiRNA--CtU6.F represents relative change of miRNA expression compared to U6 in the same sample.SPSS 19.0 was used as statistical software; t test was used for comparison of two sets of samples One way ANOVA was used for multiple groups' comparison,and make multiple comparison by the S-N-K method if the result with a significant difference.Pearson correlation analysis were used for the relationship between two variables,Brown-Forsythe test was used for Ct value equality testing among multiple samples.P ＜ 0.05were regarded as statistically significant.Results miR-20a (F =271.64,P ＜ 0.01),miR-21 (F =2232.51,P＜0.01),miR-205 (F=45.13,P＜0.01),miR-29a (F=19.98,P ＜0.01),miR-25 (F=313.19,P ＜ 0.01) and miR-126 (F =32.38,P ＜ 0.01) were differently expressed in the serum of lung cancer patients and healthy control or benign disease control.miR-29a,miR-25,miR-126 was down regulated in the development of malignant lung disease; miR-31 elevated in lung cancer compared with healthy control,while miR-145 fell; miR-31 expression changed with various differentiation of lung cancer (F =5.22,P ＜ 0.01)itwas significantly increased in the moderate-differentiated cancer,but decreased when distant metastasis existed (especially bone metastasis).But in PBMC paired with the serum samples above,statistically significance was shown in lung cancer and healthy control group and benign lung diseases group in miR-126 (F=690.58,P＜0.01),miR-129 (F=26.66,P＜0.01),miR-145 (F=48.57,P＜0.01),miR-205 (F=308.61,P＜0.01).miR-25 (F=218.57,P＜0.01) and miR-31 (F=48.05,P＜0.01),were down regulated in the development of malignant lung disease.miR-20a,miR-29a were elevated in lung cancer compared to healthy controls,miR-21 was up-regulated when distant metastasis existed; the expression of miR-31 in serum and PBMC was negatively correlated (r =-0.369,P ＜ 0.05).Areas under ROC curve of miR-25 (S =0.906,P ＜ 0.01) and miR-126 (S =0.969,P ＜ 0.01) were statistically different.Conclusions miRNA may contribute to several steps of metastasis,including local invasion,extravasation or initial survival at a distant site,and metastatic colonization,or can affect the prognosis of lung cancer.The detection of miRNA in lung cancer provides a new clue to the research of its chronic progress.