Abstract
Lymph node (LN) metastasis is often an early event in the progression of malignant tumors and it contributes to the majority of cancer mortalities. MiRNAs play key roles in tumor metastasis. This study aimed to investigate the specific miRNAs as putative indicators of metastasis early diagnosis for non-small cell lung cancer (NSCLC). In this study, five NSCLC cases with LN metastasis and four cases without metastasis (NLN) were enrolled for Agilent Human miRNA array. The interested differentially expressed miRNA was validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in the LN metastasis (n = 46) and NLN (n = 39) groups. The microarray results revealed that three miRNAs (miR-1260b, miR-423-3p, miR-23a-5p) were differentially expressed in LN metastasis group compared with NLN group. The expression of miR-1260b was tested by qRT-PCR and the mean relative expression fold change (2(-ΔΔCt)) in LN metastasis was significantly higher than that in the NLN group (3.942, 1.743 respectively, P = 1.179E-04). The patients with tumor-node-metastasis (TNM) stage III were identified more frequently in LN metastasis group (P = 1.772E-11) and with a higher expression level of miR-1260b (5.126, P = 1.147E-06). In addition, the LN metastasis cases were associated with a poorly differentiated degree (P = 0.007). The overexpression of miR-1260b in NSCLC with LN metastasis can be regarded as a specific signature for early progression and prognosis of NSCLC.
Highlights
Lung cancer is currently the leading cause of death worldwide and 80% of patients are diagnosed as nonsmall cell lung cancer (NSCLC) [1,2]
Fresh frozen tissues obtained from five NSCLC cases with Lymph node (LN) metastasis, four cases without LN (NLN) metastasis, and their paired para-carcinoma tissues were used for miRNA array analysis
After removing the reduplicative miRNAs, we analyzed the 51-pooled miRNAs for the comparison of the LN metastasis and NLN groups, and found 3 differentially expressed miRNAs (Fig. 1). Among these altered miRNAs, miR1260b was overexpressed while miR-23a-5p and miR423-3p were underexpressed in the LN metastasis group (Table 1)
Summary
Lung cancer is currently the leading cause of death worldwide and 80% of patients are diagnosed as nonsmall cell lung cancer (NSCLC) [1,2]. MicroRNAs (miRNAs) are a large family of posttranscriptional regulators of gene expression with a length of 19-22 nucleotides [6,7]. Several miRNAs such as miR-145, miR-186 and the let-7 family were considered suppressors for the metastasis and epithelial mesenchymal transition (EMT) in lung cancer [2, 9, 10] while miR-10b and miR-155 were positively correlated with the NSCLC tumorous process [11, 12]. MiR-1260b and non-small cell lung cancer to identify differentially expressed miRNAs in NSCLC with LN metastasis through miRNA array and quantitative reverse transcription polymerase chain reaction (qRT-PCR). The obtained raw data were normalized by Quantile algorithm with Gene Spring Software 12.6 (Agilent technologies, Santa Clara, CA, US)
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