Monitoring microRNAs Using a Molecular Beacon in CD133+/CD338+ Human Lung Adenocarcinoma-initiating A549 Cells

  • Yao, Quan (Diagnosis and Treatment Center of Cancer, Chengdu Military General Hospital) ;
  • Sun, Jian-Guo (Institute of Cancer, Xinqiao Hospital, Third Military Medical University) ;
  • Ma, Hu (Department of Oncology, Affiliated Hospital of Zunyi Medical University) ;
  • Zhang, An-Mei (Institute of Cancer, Xinqiao Hospital, Third Military Medical University) ;
  • Lin, Sheng (Department of Oncology, Affiliated Hospital of Luzhou Medical College) ;
  • Zhu, Cong-Hui (Institute of Cancer, Xinqiao Hospital, Third Military Medical University) ;
  • Zhang, Tao (Diagnosis and Treatment Center of Cancer, Chengdu Military General Hospital) ;
  • Chen, Zheng-Tang (Institute of Cancer, Xinqiao Hospital, Third Military Medical University)
  • Published : 2014.01.15


Lung cancer is the most common causes of cancer-related deaths worldwide, and a lack of effective methods for early diagnosis has greatly impacted the prognosis and survival rates of the affected patients. Tumor-initiating cells (TICs) are considered to be largely responsible for tumor genesis, resistance to tumor therapy, metastasis, and recurrence. In addition to representing a good potential treatment target, TICs can provide clues for the early diagnosis of cancer. MicroRNA (miRNA) alterations are known to be involved in the initiation and progression of human cancer, and the detection of related miRNAs in TICs is an important strategy for lung cancer early diagnosis. As Hsa-miR-155 (miR-155) can be used as a diagnostic marker for non-small cell lung cancer (NSCLC), a smart molecular beacon of miR-155 was designed to image the expression of miR-155 in NSCLC cases. TICs expressing CD133 and CD338 were obtained from A549 cells by applying an immune magnetic bead isolation system, and miR-155 was detected using laser-scanning confocal microscopy. We found that intracellular miR-155 could be successfully detected using smart miR-155 molecular beacons. Expression was higher in TICs than in A549 cells, indicating that miR-155 may play an important role in regulating bio-behavior of TICs. As a non-invasive approach, molecular beacons could be implemented with molecular imaging to diagnose lung cancer at early stages.


  1. Zandberga E, Kozirovskis V, Abols A, et al (2013). Cell-free microRNAs as diagnostic, prognostic, and predictive biomarkers for lung cancer. Genes Chromosomes Cancer, 52, 356-69.
  2. Zhou S, Schuetz JD, Bunting KD, et al (2001). The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the sidepopulation phenotype. Nat Med, 7, 1028-34.
  3. Wu K, Ding J, Chen C, et al (2012). Hepatic transforming growth factor beta gives rise to tumor-initiating cells and promotes liver cancer development. Hepatology, 56, 2255-67.
  4. Tyagi S, Kramer FR (1996). Molecular beacons: probes that fluoresce upon hybridization. Nat Biotechnol, 14, 303-8.
  5. Wang B, Majumder S, Nuovo G, et al (2009a). Role of microRNA-155 at early stages of hepatocarcinogenesis induced by choline-deficient and amino acid-defined diet in C57BL/6 mice. Hepatology, 50, 1152-61.
  6. Wang QZ, Xu W, Habib N, Xu R (2009b). Potential uses of microRNA in lung cancer diagnosis, prognosis, and therapy. Curr Cancer Drug Targets, 9, 572-94.
  7. Xie Y, Todd NW, Liu Z, et al (2010). Altered miRNA expression in sputum for diagnosis of non-small cell lung cancer. Lung Cancer, 67, 170-6.
  8. Yanaihara N, Caplen N, Bowman E, et al (2006). Unique microRNA molecular profiles in lung cancer diagnosis and prognosis. Cancer Cell, 9, 189-98.
  9. Yao Q, Zhang AM, Ma H, et al (2012). Novel molecular beacons to monitor microRNAs in non-small-cell lung cancer. Mol Cell Probes, 26, 182-7.
  10. Yin S, Li J, Hu C, et al (2007). CD133 positive hepatocellular carcinoma cells possess high capacity for tumorigenicity. Int J Cancer, 120, 1444-50.
  11. Yu F, Deng H, Yao H, et al (2010). Mir-30 reduction maintains self-renewal and inhibits apoptosis in breast tumor-initiating cells. Oncogene, 29, 4194-204.
  12. Yu F, Jiao Y, Zhu Y, et al (2012). MicroRNA 34c gene downregulation via DNA methylation promotes self-renewal and epithelial-mesenchymal transition in breast tumor-initiating cells. J Biol Chem, 287, 465-73.
  13. Ryu JK, Hong SM, Karikari CA, et al (2010). Aberrant MicroRNA-155 expression is an early event in the multistep progression of pancreatic adenocarcinoma. Pancreatology, 10, 66-73.
  14. Meng X, Li M, Wang X, Wang Y, Ma D (2009). Both CD133+ and CD133-subpopulations of A549 and H446 cells contain cancer-initiating cells. Cancer Sci, 100, 1040-6.
  15. O'Brien CA, Pollett A, Gallinger S, Dick JE (2007). A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature, 445, 106-10.
  16. Ricci-Vitiani L, Lombardi DG, Pilozzi E, et al (2007). Identification and expansion of human colon-cancerinitiating cells. Nature, 445, 111-5.
  17. Santangelo P, Nitin N, Bao G (2006). Nanostructured probes for RNA detection in living cells. Ann Biomed Eng, 34, 39-50.
  18. Seo DC, Sung JM, Cho HJ, et al (2007). Gene expression profiling of cancer stem cell in human lung adenocarcinoma A549 cells. Mol Cancer, 6, 75.
  19. Singh SK, Clarke ID, Terasaki M, et al (2003). Identification of a cancer stem cell in human brain tumors. Cancer Res, 63, 5821-8.
  20. Singh SK, Hawkins C, Clarke ID, et al (2004). Identification of human brain tumour initiating cells. Nature, 432, 396-401.
  21. Suetsugu A, Nagaki M, Aoki H, et al (2006). Characterization of CD133+ hepatocellular carcinoma cells as cancer stem/ progenitor cells. Biochem Biophys Res Commun, 351, 820-824.
  22. Sullivan JP, Minna JD, Shay JW (2010). Evidence for selfrenewing lung cancer stem cells and their implications in tumor initiation, progression, and targeted therapy. Cancer Metastasis Rev, 29, 61-72.
  23. Tyagi S, Bratu DP, Kramer FR (1998). Multicolor molecular beacons for allele discrimination. Nat Biotechnol, 16, 49-53.
  24. Jemal A, Siegel R, Ward E, et al (2006). Cancer statistics, 2006. CA Cancer J Clin, 56, 106-30.
  25. Henschke CI, Yankelevitz DF, Libby DM, et al (2006). Survival of patients with stage I lung cancer detected on CT screening. N Engl J Med, 355, 1763-71.
  26. Hermann PC, Huber SL, Herrler T, et al (2007). Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell, 1, 313-23.
  27. Ho MM, Ng AV, Lam S, Hung JY (2007). Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res, 67, 4827-33.
  28. Jiang S, Zhang HW, Lu MH, et al (2010). MicroRNA-155 functions as an OncomiR in breast cancer by targeting the suppressor of cytokine signaling 1 gene. Cancer Res, 70, 3119-27.
  29. Juan D, Alexe G, Antes T, et al (2010). Identification of a microRNA panel for clear-cell kidney cancer. Urology, 75, 835-41.
  30. Kim Y, Yang CJ, Tan W (2007). Superior structure stability and selectivity of hairpin nucleic acid probes with an L-DNA stem. Nucleic Acids Res, 35, 7279-87.
  31. Kong W, He L, Coppola M, et al (2010). MicroRNA-155 regulates cell survival, growth, and chemosensitivity by targeting FOXO3a in breast cancer. J Biol Chem, 285, 17869-79.
  32. Krol J, Loedige I, Filipowicz W (2010). The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet, 11, 597-610.
  33. Kwak PB, Iwasaki S, Tomari Y (2010). The microRNA pathway and cancer. Cancer Sci, 101, 2309-15.
  34. Kwon MJ, Shin YK (2013). Regulation of ovarian cancer stem cells or tumor-initiating cells. Int J Mol Sci, 14, 6624-48.
  35. Garofalo M, Croce CM (2011). microRNAs: Master regulators as potential therapeutics in cancer. Annu Rev Pharmacol Toxicol, 51, 25-43.
  36. Elton TS, Selemon H, Elton SM, Parinandi NL (2012). Regulation of the MIR155 host gene in physiological and pathological processes. Gene, 532, 1-12
  37. Eramo A, Lotti F, Sette G, et al (2008). Identification and expansion of the tumorigenic lung cancer stem cell population. Cell Death Differ, 15, 504-14.
  38. Faraoni I, Antonetti FR, Cardone J, Bonmassar E (2009). miR-155 gene: a typical multifunctional microRNA. Biochim Biophys Acta, 1792, 497-505.
  39. Adhikari AS, Agarwal N, Iwakuma T (2011). Metastatic potential of tumor-initiating cells in solid tumors. Front Biosci, 16, 1927-38.
  40. Bartel DP (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Cell, 116, 281-97.
  41. Garzon R, Calin GA, Croce CM (2009). MicroRNAs in Cancer. Annu Rev Med, 60, 167-79.
  42. Georgantas RW, Hildreth R, Morisot S, et al (2007). CD34+ hematopoietic stem-progenitor cell microRNA expression and function: a circuit diagram of differentiation control. Proc Natl Acad Sci USA, 104, 2750-5.
  43. Greither T, Grochola LF, Udelnow A, et al (2010). Elevated expression of microRNAs 155, 203, 210 and 222 in pancreatic tumors is associated with poorer survival. Int J Cancer, 126, 73-80.
  44. Guo W, Lasky JL, Wu H (2006). Cancer stem cells. Pediatr Res, 59, 59-64.
  45. Habbe N, Koorstra JB, Mendell JT, et al (2009). MicroRNA miR-155 is a biomarker of early pancreatic neoplasia. Cancer Biol Ther, 8, 340-6.
  46. Hatfield S, Ruohola-Baker H (2008). microRNA and stem cell function. Cell Tissue Res, 331, 57-66.
  47. Bertolini G, Roz L, Perego P, et al (2009). Highly tumorigenic lung cancer CD133+ cells display stem-like features and are spared by cisplatin treatment. Proc Natl Acad Sci USA, 106, 16281-6.
  48. Bushati N, Cohen SM (2007). microRNA functions. Annu Rev Cell Dev Biol, 23, 175-205.
  49. Chen CZ (2005). MicroRNAs as oncogenes and tumor suppressors. N Engl J Med, 353, 1768-71.
  50. Collins AT, Berry PA, Hyde C, Stower MJ, Maitland NJ (2005). Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res, 65, 10946-51.
  51. Donnem T, Eklo K, Berg T, et al (2011). Prognostic impact of MiR-155 in non-small cell lung cancer evaluated by in situ hybridization. J Transl Med, 9, 6.

Cited by

  1. Advances in the Early Detection of Lung Cancer using Analysis of Volatile Organic Compounds: From Imaging to Sensors vol.15, pp.11, 2014,
  2. miR-421, miR-155 and miR-650: Emerging Trends of Regulation of Cancer and Apoptosis vol.15, pp.5, 2014,
  3. Matrine Reduces Proliferation of Human Lung Cancer Cells by Inducing Apoptosis and Changing miRNA Expression Profiles vol.15, pp.5, 2014,
  4. Chitosan Combined with Molecular Beacon for Mir-155 Detection and Imaging in Lung Cancer vol.19, pp.9, 2014,
  5. Effect of CXCR4 and CD133 Co-expression on the Prognosis of Patients with Stage II~III Colon Cancer vol.16, pp.3, 2015,
  6. Targeting the Wnt-Regulatory Protein CTNNBIP1 by microRNA-214 Enhances the Stemness and Self-Renewal of Cancer Stem-Like Cells in Lung Adenocarcinomas vol.33, pp.12, 2015,
  7. Prognostic value of cancer stem cell marker CD133 expression in esophageal carcinoma: A meta-analysis vol.4, pp.1, 2015,