A three-microRNA signature for lung squamous cell carcinoma diagnosis in Chinese male patients

Various studies have demonstrated the diagnostic value of microRNA (miRNA) for lung cancer, but miRNA signatures varied between different subtypes. Whether serum miRNAs could be used as biomarkers in lung squamous cell carcinoma (SCC) remains unknown. Using quantitative real-time polymerase chain reaction (qRT-PCR) based Exiqon panel, 38 differentially expressed miRNAs were identified from 3 male lung SCC pool samples and 1 normal control (NC) pool in the initial screening phase. After the training (24 SCC VS. 15 NCs), testing (44 SCC VS. 57 NCs) and external validation (34 SCC VS. 36 NCs VS. 10 pulmonary hamartoma) processes via qRT-PCR, we identified a three-miRNA panel ((miR-106a-5p, miR-20a-5p and miR-93-5p) to be a potential diagnostic marker for male lung SCC patients. The areas under the receiver operating characteristic (ROC) curve of the three-miRNA panel for the training, testing and validation phases were 0.969, 0.881 and 0.954 respectively. In addition, this signature could also differentiate lung SCC from pulmonary hamartoma (AUC=0.900). The 3 miRNAs were consistently up-regulated in lung SCC tissues (23 SCC VS. 23 NCs) and serum exosomes (17 SCC VS. 24 NCs). Moreover, expression of the 3 miRNAs was decreased in arterial serum (n = 3). In conclusion, we established a three-miRNA signature in the peripheral serum with considerable clinical value in the diagnosis of male lung SCC patients.

[1]  John D. Minna,et al.  miR-93, miR-98, and miR-197 Regulate Expression of Tumor Suppressor Gene FUS1 , 2009, Molecular Cancer Research.

[2]  Jiang Li,et al.  Tumor-derived exosomes promote tumor progression and T-cell dysfunction through the regulation of enriched exosomal microRNAs in human nasopharyngeal carcinoma , 2014, Oncotarget.

[3]  Xian-gao Jiang,et al.  ZNRF3 contributes to the growth of lung carcinoma via inhibiting Wnt/β-catenin pathway and is regulated by miR-93 , 2016, Tumor Biology.

[4]  X. Liu,et al.  Expression of miR-29c, miR-93, and miR-429 as Potential Biomarkers for Detection of Early Stage Non-Small Lung Cancer , 2014, PloS one.

[5]  E. Brambilla,et al.  The evolving role of histology in the management of advanced non-small-cell lung cancer. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[6]  X. Su,et al.  MicroRNAs May Serve as Emerging Molecular Biomarkers for Diagnosis and Prognostic Assessment or as Targets for Therapy in Gastric Cancer. , 2015, Asian Pacific journal of cancer prevention : APJCP.

[7]  M. Mahdipour,et al.  Validating reference microRNAs for normalizing qRT-PCR data in bovine oocytes and preimplantation embryos , 2015, BMC Developmental Biology.

[8]  Holger Sültmann,et al.  Circulating miRNAs are correlated with tumor progression in prostate cancer , 2011, International journal of cancer.

[9]  Xi Chen,et al.  Serum microRNA expression profile as a biomarker in the diagnosis and prognosis of pancreatic cancer. , 2012, Clinical chemistry.

[10]  Y. Shu,et al.  MiR-4728-3p could act as a marker of HER2 status. , 2015, Cancer biomarkers : section A of Disease markers.

[11]  Guangxue Wang,et al.  A direct quantification method for measuring plasma MicroRNAs identified potential biomarkers for detecting metastatic breast cancer , 2016, Oncotarget.

[12]  Christina Backes,et al.  High-throughput qRT-PCR validation of blood microRNAs in non-small cell lung cancer , 2015, Oncotarget.

[13]  A. McCullough.  Comprehensive genomic characterization of squamous cell lung cancers , 2013 .

[14]  B. Su,et al.  Identification of serum miRNAs by nano-quantum dots microarray as diagnostic biomarkers for early detection of non-small cell lung cancer , 2016, Tumor Biology.

[15]  Xin-shu Dong,et al.  The potential of plasma miRNAs for diagnosis and risk estimation of colorectal cancer. , 2015, International journal of clinical and experimental pathology.

[16]  Chuan Wang,et al.  Serum microRNA panel as biomarkers for early diagnosis of colorectal adenocarcinoma , 2014, British Journal of Cancer.

[17]  Daniel B. Martin,et al.  Circulating microRNAs as stable blood-based markers for cancer detection , 2008, Proceedings of the National Academy of Sciences.

[18]  R. Biondi,et al.  Differential Stability of Cell-Free Circulating microRNAs: Implications for Their Utilization as Biomarkers , 2013, PloS one.

[19]  P. Buckhaults,et al.  An epigenetic auto-feedback loop regulates TGF-β type II receptor expression and function in NSCLC , 2015, Oncotarget.

[20]  Nicola J. Armstrong,et al.  Haemolysis during Sample Preparation Alters microRNA Content of Plasma , 2011, PloS one.

[21]  L. Tanoue,et al.  New TNM classification for non-small-cell lung cancer , 2009, Expert review of anticancer therapy.

[22]  Q. Lan,et al.  Identification of 9 serum microRNAs as potential noninvasive biomarkers of human astrocytoma. , 2015, Neuro-oncology.

[23]  A. Jemal,et al.  Global cancer statistics, 2012 , 2015, CA: a cancer journal for clinicians.

[24]  R. Askeland,et al.  MiR-93 enhances angiogenesis and metastasis by targeting LATS2 , 2012, Cell cycle.

[25]  A Shiotani,et al.  Identification of serum miRNAs as novel non-invasive biomarkers for detection of high risk for early gastric cancer , 2013, British Journal of Cancer.

[26]  Wei Wang,et al.  Utility of squamous cell carcinoma antigen, carcinoembryonic antigen, Cyfra 21-1 and neuron specific enolase in lung cancer diagnosis: a prospective study from China. , 2011, Chinese medical journal.

[27]  N. Girard,et al.  Comparisons of microRNA Patterns in Plasma before and after Tumor Removal Reveal New Biomarkers of Lung Squamous Cell Carcinoma , 2013, PloS one.

[28]  Miao Zhang,et al.  Diagnostic significance of miR-106a in gastric cancer. , 2015, International journal of clinical and experimental pathology.

[29]  J. Minna,et al.  miR-93-directed down-regulation of DAB2 defines a novel oncogenic pathway in lung cancer , 2013, Oncogene.

[30]  D. Wheeler,et al.  Identification of a pan-cancer oncogenic microRNA superfamily anchored by a central core seed motif , 2013, Nature Communications.

[31]  Xue-mei Ma,et al.  Identification of Suitable Reference Genes for qPCR Analysis of Serum microRNA in Gastric Cancer Patients , 2012, Digestive Diseases and Sciences.

[32]  J. Nikliński,et al.  Validation for histology‐driven diagnosis in non‐small cell lung cancer using hsa‐miR‐205 and hsa‐miR‐21 expression by two different normalization strategies , 2016, International journal of cancer.

[33]  N. Saavedra,et al.  Identification of microRNAs involved in the modulation of pro-angiogenic factors in atherosclerosis by a polyphenol-rich extract from propolis. , 2014, Archives of biochemistry and biophysics.

[34]  Yanhong Zhou,et al.  Circulating miR-17, miR-20a, miR-29c, and miR-223 Combined as Non-Invasive Biomarkers in Nasopharyngeal Carcinoma , 2012, PloS one.

[35]  A. Schetter,et al.  Circulating micro‐RNA expression profiles in early stage nonsmall cell lung cancer , 2012, International journal of cancer.

[36]  A. Bahnassy,et al.  Circulating Serum miRNAs as Diagnostic Markers for Colorectal Cancer , 2016, PloS one.

[37]  Yaming Li,et al.  MicroRNA-106a confers cisplatin resistance in non-small cell lung cancer A549 cells by targeting adenosine triphosphatase-binding cassette A1. , 2015, Molecular medicine reports.

[38]  Rui Hu,et al.  miR-106a promotes growth and metastasis of non-small cell lung cancer by targeting PTEN. , 2015, International journal of clinical and experimental pathology.

[39]  Jian Wang,et al.  MiR‐145, miR‐133a and miR‐133b inhibit proliferation, migration, invasion and cell cycle progression via targeting transcription factor Sp1 in gastric cancer , 2014, FEBS letters.

[40]  Siqi Liu,et al.  Identification and validation of rice reference proteins for western blotting , 2011, Journal of experimental botany.

[41]  Q. Shan,et al.  Serum miR-210 and miR-30a expressions tend to revert to fetal levels in Chinese adult patients with chronic heart failure. , 2013, Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology.

[42]  Fang Wang,et al.  Diagnostic value of a plasma microRNA signature in gastric cancer: a microRNA expression analysis , 2015, Scientific Reports.

[43]  D. Beer,et al.  A Novel Serum 4-microRNA Signature for Lung Cancer Detection , 2015, Scientific Reports.

[44]  Jia’nan Xie,et al.  Identification and validation of reference genes for quantitative RT-PCR analysis of retinal pigment epithelium cells under hypoxia and/or hyperglycemia. , 2016, Gene.

[45]  Wei Li,et al.  Identification and Validation of Reference Genes for qPCR Detection of Serum microRNAs in Colorectal Adenocarcinoma Patients , 2013, PloS one.

[46]  L. Hood,et al.  Circulating microRNAs, potential biomarkers for drug-induced liver injury , 2009, Proceedings of the National Academy of Sciences.

[47]  K. Pantel,et al.  Diagnostic and prognostic potential of serum miR-7, miR-16, miR-25, miR-93, miR-182, miR-376a and miR-429 in ovarian cancer patients , 2015, British Journal of Cancer.

[48]  P. Bonomi,et al.  Treatment of advanced squamous cell carcinoma of the lung: a review. , 2015, Translational lung cancer research.

[49]  Zhaoyong Li,et al.  A plasma microRNA panel for early detection of colorectal cancer , 2015, International journal of cancer.

[50]  F. He,et al.  Expression of circulating microRNA-20a and let-7a in esophageal squamous cell carcinoma. , 2015, World journal of gastroenterology.

[51]  E. Lianidou,et al.  Quantification of circulating miRNAs in plasma: effect of preanalytical and analytical parameters on their isolation and stability. , 2013, The Journal of molecular diagnostics : JMD.

[52]  Tao Jiang,et al.  Circulating microRNAs in cancer: origin, function and application , 2012, Journal of Experimental & Clinical Cancer Research.