Skip to main content
SpringerLink
Log in

Aberrant microRNAs expression in CD133+/CD326+ human lung adenocarcinoma initiating cells from A549

  • Published:
Molecules and Cells

Abstract

Increasing evidence demonstrates that miRNAs are involved in the dysregulation of tumor initiating cells (TICs) in various tumors. Due to a lack of definitive markers, cell sorting is not an ideal separation method for lung adenocarcinoma initiating cells. In this study, we combined paclitaxel with serum-free medium cultivation (inverse-induction) to enrich TICs from A549 cells, marked by CD133/CD326, defined features of stemness. We next investigated aberrant microRNAs in this subpopulation compared to normal cells with miRNA microarray and found that 50 miRNAs exhibited a greater than 2-fold change in expression. As further validation, 10 miRNAs were chosen to perform quantitative RT-PCR on the A549 cell line and primary samples. The results suggest that aberrant expression of miRNAs such as miR-29ab, miR-183, miR-17-5p and miR-127-3P may play an important role in regulating the bio-behavior of TICs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Al-Hajj, M., Wicha, M.S., Hernandez, A.B., Morrison, S.J., and Clarke, M.F. (2003). Prospective identification of tumorigenic breast cancer cells. Proc. Natl. Acad. Sci. USA 100, 3983–3988.

    Article  PubMed  CAS  Google Scholar 

  • Bonnet, D., and Dick, J.E. (1997). Human acute myeloid leukemia is organized as a hierarchy that originates form a primitive hematopoietic cell. Nat. Med. 3, 730–737.

    Article  PubMed  CAS  Google Scholar 

  • Bertolinia, G., Roza, L., Peregob, P., Tortoretob, M., Fontanellac, E., Gattib, L., Pratesib, G., Fabbrid, A., Andriania, F., Tinellib, S., 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, 16281–16286.

    Article  Google Scholar 

  • Chiou, S.H., Yu, C.C., Huang, C.Y., Lin, S.C., Liu, C.J., Tsai, T.H., Chou, S.H., Chien, C.S., Ku, H.H., and Lo, J.F. (2008). Positive correlations of Oct-4 and Nanog in oral cancer stem-like cells and high-grade oral squamous cell carcinoma. Clin. Cancer Res. 14, 4085–4095.

    Article  PubMed  CAS  Google Scholar 

  • Cloonan, N., Brown, M.K., Steptoe, A.L., Wani, S., Chan, W.L., Forrest, A.R.R., Kolle, G., Gabrielli, B., and Grimmond, S.M. (2008). The miR-17-5p microRNA is a key regulator of the G1/S phase cell cycle transition. Genome Biol. 9, R127.

    Article  PubMed  Google Scholar 

  • Collins, A.T., and Maitland, N.J. (2006). Prostate cancer stem cells. Eur. J. Cancer 42 1213–1218.

    Article  PubMed  CAS  Google Scholar 

  • DeSano, J.T., and Xu, L. (2009). MicroRNA regulation of cancer stem cells and therapeutic implications. AAPS J. 11, 682–692.

    Article  PubMed  CAS  Google Scholar 

  • Eramo, A., Lotti, F., Sette, G., Pilozzi, E., Biffoni, M., Virgilio, A.D., Conticello, C., Ruco, L., Peschle, C., and Maria, R.D. (2008). Identification and expansion of the tumorigenic lung cancer stem cell population. Cell Death Differ.15, 504–514.

    Article  PubMed  CAS  Google Scholar 

  • Garzia, L., Andolfo, I., Cusanelli, E., Marino, N., Petrosino, G., Martino, D.D., Esposito, V., Galeone, A., Navas, L., Esposito, S., et al. (2009). MicroRNA-199b-5p impairs cancer stem cells through negative regulation of HES1 in medulloblastoma. PLOS One 4, e–4998.

    Article  Google Scholar 

  • Godlewskil, J., Nowicki, M.O., Bronisz, A., Williams, S., Otsuki, A., Nuovo, G., Chaudhury, A.R., Newton, H.B., Chiocca, E.A., and Lawler, S. (2008). Targeting of the Bmi-1 oncogene/stem cell renewal factor by microRNA-128 inhibits glioma proliferation and self-renewal. Cancer Res. 68, 9125–9130.

    Article  Google Scholar 

  • Hatfield, S., and Ruohola-Baker, H. (2008).microRNA and stem cell function. Cell Tissue Res. 331, 57–66.

    Article  PubMed  CAS  Google Scholar 

  • Hermann, P.C., Huber, S.L., Herrler, T., Aicher, A., Ellwart, J.W., Guba, M., Bruns, C.J., and Heeschen, C. (2007). Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 1, 313–323.

    Article  PubMed  CAS  Google Scholar 

  • Hossain, A., Kuo, M.T., and Saunders, G.F. (2006). Mir-17-5p regulates breast cancer cell proliferation by inhibiting translation of AIB1 mRNA. Mol. Cell. Biol. 26, 8191–8201.

    Article  PubMed  CAS  Google Scholar 

  • Ji, J., Yamashita, T., Budhu, A., Forgues, M., Jia, H.L., Li, C., Deng, C., Wauthier, E., Reid, L.M., Ye, Q.H., et al. (2009). Identification of microRNA-181 by genome-wide screening as a critical player in EpCAM-positive hepatic cancer stem cells. Hepatology 50, 472–480.

    Article  PubMed  CAS  Google Scholar 

  • Ji, Q., Hao, X., Zhang, M., Tang, W., Meng, Y., Li, L., Xiang, D., Jeffrey, T., Sano, D., Bommer, G.T., et al. (2009). MicroRNA miR-34 inhibits human pancreatic cancer tumor-initiating cells. PLOS One 4, e6816.

    Article  PubMed  Google Scholar 

  • Jiang, J., Gusev, Y., Aderca, I., Mettler, T.A., Nagorney, D.M., Brackett, D.J., Roberts, L.R., and Schmittgen, T.D. (2008). Association of MicroRNA expression in hepatocellular carcinomas with hepatitis infection, cirrhosis, and patient survival. Clin. Cancer Res. 14, 419–427.

    Article  PubMed  CAS  Google Scholar 

  • Levina, V., Marrangoni, A.M., DeMarco, R., Gorelik, E., and Lokshin, A.E. (2008). Drug-elected human lung cancer stem cells: cytokine network, tumorigenic and metastatic properties. PLOS One 3, e3077.

    Article  PubMed  Google Scholar 

  • Li, G., Luna, C., Qiu, J., Epstein, D.L., and Gonzaenz, P. (2010). Targeting of integrinβ1 and kinesin 2A by microRNA 183. J. Biol. Chem. 285, 5461–5471.

    Article  PubMed  CAS  Google Scholar 

  • Li, H., Bian, C., Liao, L., Li, J., and Zhao, R.C. (2011). miR-17-5p promotes human breast cancer cell migration and invasion through suppression HBP1. Breast Cancer Res. Treat. 126. 565–575.

    Article  PubMed  CAS  Google Scholar 

  • Li, J., Fu, H., Xu, C., Tie, Y., Xing, R., Zhu, J., Qin, Y., Sun, Z., and Zheng, X. (2011). MiR-183 inhibits TGFβ1-induced apoptosis by downregulation of PDCD4 expression in human hepatocellular carcinoma cells. BMC Cancer 10, 354.

    Article  Google Scholar 

  • Lu, W., Thomson, J.M., Wong, H.Y.F., Hammond, S.M., and Hogan, B.L.M. (2007). Transgenic over-expression of the microRAN miR-17-92 cluster promotes proliferation and inhibits differenttiation of lung epithelial progenitor cells. Dev. Biol. 310, 442–453.

    Article  PubMed  CAS  Google Scholar 

  • Ma, S., Tang, K., Chan, Y., Lee, T., Kwan, P., Castilho, A., Ng, I., Man, K., Wong, N., To, K.F., et al. (2010). miR-130b promotes CD133+ liver tumor-initiating cell growth and self-renewal via tumor protein 53-induced nuclear protein 1. Cell Stem Cell 7, 694–707.

    Article  PubMed  CAS  Google Scholar 

  • Meng, X., Li, M., Wang, X., Wang, Y., and Ma, D. (2009). Both CD133+ and CD133− sub populations of A549 and H446 cells contain cancer-initiating cells. Cancer Sci. 100, 1040–1046.

    Article  PubMed  CAS  Google Scholar 

  • Peacock, D.C., and Watkins, D.N. (2008). Cancer stem cells and the ontogeny of lung cancer. J. Clin. Oncol. 26, 2883–2889.

    Article  PubMed  CAS  Google Scholar 

  • Ricci-Vitiani, L., Lombardi, D.G., Pilozzi, E., Biffoni, M., Todaro, M., Peschle, C., and Maria, R.D. (2007). Identification and expansion of human colon-cancer-initiating cells. Nature 445, 111–115.

    Article  PubMed  CAS  Google Scholar 

  • Saito, Y., Liang, G., Egger, G., Friedman, J.M., Chuang, J., Coetzee, G.A., and Jones, P.A. (2006). Specific activation of microRNA-127 with down-regulation of the proto-oncogene BCL6 by chromatin-modifying drugs in human cancer cells. Cancer Cell 9. 435–443.

    Article  PubMed  CAS  Google Scholar 

  • Saver, A.L., Li, L.H., and Subramanian, S. (2010). MicroRNA miR-183 functions as an oncogene by targeting the transcription factor EGR1 and promoting tumor cell migration. Cancer Res. 70, 9570–9580.

    Article  Google Scholar 

  • Singh, S.K., Hawkins, C., Clarke, I.D., Squire, J.A., Bayani, J., Hide, T., Henkelman, R.M., Cusimano, M.D., and Dirks, P.B. (2004). Identification of a cancer stem cell in human brain tumors. Nature 432, 396–401.

    Article  PubMed  CAS  Google Scholar 

  • Shan, S.W., Lee, D.Y., Deng, Z., Shatseva, T., Jeyapalan, Z., Du, W.W., Zhang, Y., Xuan, J.W., Yee, S.P., Siragam, V., et al. (2009). MicroRNA miR-17 retards tissue growth and represses fibronectin expression. Nat. Cell Biol. 11, 1031–1038.

    Article  PubMed  CAS  Google Scholar 

  • Shi, L., Zhang, J.X., Pan, T.H., Zhou, J.F., Gong, W.Y., Liu, N., Fu, Z., and You, Y.P. (2010). MiR-125b is critical for the suppression of human U251 glioma stem cell proliferation. Brain Res. 1312, 120–126.

    Article  PubMed  CAS  Google Scholar 

  • Shimono, Y., Zabala, M., Cho, R.W., Lobo, N., Dalerba, P., Qian, D., Diehn, M., Liu, H., Panula, S.P., Chiao, E., et al. (2009). Down-regulation of miRNA-200c links breast cancer stem cells with normal stem cells. Cell 138, 592–603.

    Article  PubMed  CAS  Google Scholar 

  • Tagawa, H., Karube, K., Tsuzuki, S., Ohshima, K., Seto, M., and Tagawa, H. (2007). Synergistic action of the microRNA-17 polycistron and myc in aggressive cancer development. Cancer Sci. 98, 1482–1490.

    Article  PubMed  CAS  Google Scholar 

  • Teng, Y., Wang, X.W., Wang, Y.W., and Ma, D.X. (2010). Wnt/beta-catenin signaling regulates cancer stem cells in lung cancer A549 cells. Biochem. Biophys. Res. Commun. 392, 373–379.

    Article  PubMed  CAS  Google Scholar 

  • Tirino, V., Camerlingo, R., Franco, R., Malanga, D., Rocca, A.L., Viglietto, G., Rocco, G., and Pirozzi, G. (2009). The role of CD133 in the identification and characterisation of tumour-initiating cells in non-small-cell lung cancer. Eur. J. Cardiothorac. Surg. 36, 446–453.

    Article  PubMed  Google Scholar 

  • Tryndyak, V.P., Ross, S.A., Beland, F.A., and Pogribny, I.P. (2009). Down-regulation of the microRNAs miR-34a, miR-127, and miR-200b in rat liver during hepatocarcinogenesis induced by a methy1-deficient diet. Mol. Carcinog. 48, 479–487.

    Article  PubMed  CAS  Google Scholar 

  • Ura, S., Honda, M., Yamashita, T., Ueda, T., Takatori, H., Nishino, R., Sunakozaka, H., Sakai, Y., Horimoto, K., and Kaneko, S. (2009). Differential microRNA expression between hepatitis B and hepatitis C leading disease progression to hepatocellular carcinoma. Hepatology 49, 1098–1112.

    Article  PubMed  CAS  Google Scholar 

  • Yu, F., Deng, H., Yao, H., Liu, Q., Su, F., and Song, E. (2010). Mir-30 reduction maintainsself-renewal and inhibits apoptosis in breast tumor-initiating cells. Oncogene 29, 4194–4204.

    Article  PubMed  CAS  Google Scholar 

  • Wang, G. F., Mao, W.M., and Zheng, S. (2008). MicroRNA-183 regulates Ezrin expression in lung cancer cells. FEBS Lett. 582, 3663–3668.

    Article  PubMed  CAS  Google Scholar 

  • Wang, Z.W., Li, Y.W., Ahmad, A., Azmi, A.S., Kong, D.J., Banerjee, S., and Sarkar, F.H. (2010). Targeting miRNAs involved in cancer stem cell and EMT regulation: an emerging concept in overcoming drug resistance. Drug Resist. Updat. 13 109–118.

    Article  PubMed  CAS  Google Scholar 

  • Wiemer, E.A. (2007). The role of microRNAs in cancer: no small matter. Eur. J. Cancer 43, 1529–1544.

    Article  PubMed  CAS  Google Scholar 

  • Wong, P., Iwasaki, M., Somervaille, T.C.P, Ficara, F., Carico, C., Arenold, C., Chen, C.Z., and Cleary, M. (2010). The miR-17-92 microrna polycistron regulates MLL leukemia stem cell potential by modulating p21 expression. Cancer Res. 70, 3383–3342.

    Article  Google Scholar 

  • Wellner, U., Schubert, J., Burk, U.C., Schmalhofer, O., Zhu, F., Sonntag, A., Waldvogel, B., Vannier, C., Darling, D., Hausen, A., et al. (2009).The EMT-activator ZEB1 promotes tumorigenicity by repressing stemness-inhibiting microRNAs. Nat. Cell Biol. 11, 1487–1495.

    Article  PubMed  CAS  Google Scholar 

  • Zabierowski, S.E., and Herlyn, M. (2008). Melanoma stem cells: the dark seed of melanoma. J. Clin. Oncol. 26, 2890–2894.

    Article  PubMed  Google Scholar 

  • Zhang, S., Balch, C., Chan, M.W., Lai, H.C., Matei, D., Schilder, J.M., Yan, P.S., Huang, T.H.M., and Nephew, K.P. (2008). Identification and characterization of ovarian cancer-initiating cells from primary human tumors, Cancer Res. 68, 4311–4320.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, M., Liu, Q., Mi, S., Liang, X., Zhang, Z., Su, X., Liu, J., Chen, Y., Wang, M., Zhang, Y., et al. (2011). Both miR-17-5p and miR-20a alleviate suppressive potential of myeloid-derived suppressor cells by modulating STAT3 expression. J. Immunol. 186, 4716–4724.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Cancer, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China

    Sheng Lin, Jian-guo Sun, Hai-xia Long, Cong-hui Zhu, Tong Xiang, Hu Ma, Zhong-quan Zhao, Quan Yao, An-mei Zhang, Bo Zhu & Zheng-tang Chen

  2. Oncology Department, The Affiliated Hospital of Luzhou Medical College, Sichuan, 646000, China

    Sheng Lin & Jing-bo Wu

Authors
  1. Sheng Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jian-guo Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jing-bo Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hai-xia Long
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Cong-hui Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tong Xiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hu Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Zhong-quan Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Quan Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. An-mei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Bo Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Zheng-tang Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Bo Zhu or Zheng-tang Chen.

Additional information

These authors contributed equally to this work.

About this article

Cite this article

Lin, S., Sun, Jg., Wu, Jb. et al. Aberrant microRNAs expression in CD133+/CD326+ human lung adenocarcinoma initiating cells from A549. Mol Cells 33, 277–283 (2012). https://doi.org/10.1007/s10059-012-2252-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10059-012-2252-y

Keywords

Search

Navigation