Skip to main content

Advertisement

SpringerLink
Log in

Identifying microRNA-mRNA regulatory network in gemcitabine-resistant cells derived from human pancreatic cancer cells

  • Research Article
  • Published:
Tumor Biology

Abstract

Pancreatic cancer is unresectable in over 80 % of patients owing to difficulty in early diagnosis. Chemotherapy is the most frequently adopted therapy for advanced pancreatic cancer. The development of drug resistance to gemcitabine (GEM), which is always used in standard chemotherapy, often results in therapeutic failure. However, the molecular mechanisms underlying the gemcitabine resistance remain unclear. Therefore, we sought to explore the microRNA-mRNA network that is associated with the development of gemcitabine resistance and to identify molecular targets for overcoming the gemcitabine resistance. By exposing SW1990 pancreatic cancer cells to long-term gemcitabine with increasing concentrations, we established a gemcitabine-resistant cell line (SW1990/GEM) with a high IC50 (the concentration needed for 50 % growth inhibition, 847.23 μM). The mRNA and microRNA expression profiles of SW1990 cells and SW1990/GEM cells were determined using RNA-seq analysis. By comparing the results in control SW1990 cells, 507 upregulated genes and 550 downregulated genes in SW1990/GEM cells were identified as differentially expressed genes correlated with gemcitabine sensitivity. Gene ontology (GO) analysis showed that the differentially expressed genes were related to diverse biological processes. The upregulated genes were mainly associated with drug response and apoptosis, and the downregulated genes were correlated with cell cycle progression and RNA splicing. Concurrently, the differentially expressed microRNAs, which are the important player in drug resistance development, were also examined in SW1990/GEM cells, and 56 differential microRNAs were identified. Additionally, the expression profiles of selected genes and microRNAs were confirmed by using Q-PCR assays. Furthermore, combining the differentially expressed microRNAs and mRNAs as well as the predicted targets for these microRNAs, a core microRNA-mRNA regulatory network was constructed, which included hub microRNAs, such as hsa-miR-643, hsa-miR-4644, hsa-miR-4650-5p, hsa-miR-4455, hsa-miR-1261, and hsa-miR-3676. The predicted targets of these hub microRNAs in the microRNA-mRNA network were also observed in the identified differential genes. As a result, a differential gene and microRNA expression pattern was constructed in gemcitabine-resistant pancreatic cancer cells. Therefore, these data may be useful for the detection and treatment of drug resistance in pancreatic cancer patients, and the microRNA-mRNA network-based analysis is expected to be more effective and provides deep insights into the molecular mechanism of drug resistance.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Siegel R, Naishadham D, Jemal A. Cancer statistics, 2013. CA Cancer J Clin. 2013;63:11–30.

    Article  PubMed  Google Scholar 

  2. Warshaw AL, Fernandez-del CC. Pancreatic carcinoma. N Engl J Med. 1992;326:455–65.

    Article  CAS  PubMed  Google Scholar 

  3. Yeo CJ, Cameron JL, Sohn TA, Lillemoe KD, Pitt HA, Talamini MA, et al. Six hundred fifty consecutive pancreaticoduodenectomies in the 1990s: pathology, complications, and outcomes. Ann Surg. 1997;226:248–57.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Ahmad NA, Lewis JD, Ginsberg GG, Haller DG, Morris JB, Williams NN, et al. Long term survival after pancreatic resection for pancreatic adenocarcinoma. Am J Gastroenterol. 2001;96:2609–15.

    Article  CAS  PubMed  Google Scholar 

  5. Stefanska B, Arciemiuk M, Bontemps-Gracz MM, Dzieduszycka M, Kupiec A, Martelli S, et al. Synthesis and biological evaluation of 2,7-Dihydro-3H-dibenzo[de, h]cinnoline-3,7-dione derivatives, a novel group of anticancer agents active on a multidrug resistant cell line. Bioorg Med Chem. 2003;11:561–72.

    Article  CAS  PubMed  Google Scholar 

  6. Garcia-Manteiga J, Molina-Arcas M, Casado FJ, Mazo A, Pastor-Anglada M. Nucleoside transporter profiles in human pancreatic cancer cells: role of hCNT1 in 2′,2′-difluorodeoxycytidine-induced cytotoxicity. Clin Cancer Res. 2003;9:5000–8.

    CAS  PubMed  Google Scholar 

  7. Saif MW. Options for the treatment of gemcitabine-resistant advanced pancreatic cancer: are we there yet? JOP. 2010;11:288–9.

    PubMed  Google Scholar 

  8. Duxbury MS, Ito H, Zinner MJ, Ashley SW, Whang EE. RNA interference targeting the M2 subunit of ribonucleotide reductase enhances pancreatic adenocarcinoma chemosensitivity to gemcitabine. Oncogene. 2004;23:1539–48.

    Article  CAS  PubMed  Google Scholar 

  9. Giovannetti E, Del TM, Mey V, Funel N, Nannizzi S, Ricci S, et al. Transcription analysis of human equilibrative nucleoside transporter-1 predicts survival in pancreas cancer patients treated with gemcitabine. Cancer Res. 2006;66:3928–35.

    Article  CAS  PubMed  Google Scholar 

  10. Duxbury MS, Ito H, Zinner MJ, Ashley SW, Whang EE. Focal adhesion kinase gene silencing promotes anoikis and suppresses metastasis of human pancreatic adenocarcinoma cells. Surgery. 2004;135:555–62.

    Article  CAS  PubMed  Google Scholar 

  11. Duxbury MS, Ito H, Zinner MJ, Ashley SW, Whang EE. SiRNA directed against c-Src enhances pancreatic adenocarcinoma cell gemcitabine chemosensitivity. J Am Coll Surg. 2004;198:953–9.

    Article  PubMed  Google Scholar 

  12. Liau SS, Ashley SW, Whang EE. Lentivirus-mediated RNA interference of HMGA1 promotes chemosensitivity to gemcitabine in pancreatic adenocarcinoma. J Gastrointest Surg. 2006;10:1254–62.

    Article  PubMed  Google Scholar 

  13. Mahon PC, Baril P, Bhakta V, Chelala C, Caulee K, Harada T, et al. S100A4 contributes to the suppression of BNIP3 expression, chemoresistance, and inhibition of apoptosis in pancreatic cancer. Cancer Res. 2007;67:6786–95.

    Article  CAS  PubMed  Google Scholar 

  14. Kim MP, Gallick GE. Gemcitabine resistance in pancreatic cancer: picking the key players. Clin Cancer Res. 2008;14:1284–5.

    Article  CAS  PubMed  Google Scholar 

  15. Zhao YP, Chen G, Feng B, Zhang TP, Ma EL, Wu YD. Microarray analysis of gene expression profile of multidrug resistance in pancreatic cancer. Chin Med J (Engl). 2007;120:1743–52.

    CAS  Google Scholar 

  16. Lee KH, Galloway JF, Park J, Dvoracek CM, Dallas M, Konstantopoulos K, et al. Quantitative molecular profiling of biomarkers for pancreatic cancer with functionalized quantum dots. Nanomedicine. 2012;8:1043–51.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Carrington JC, Ambros V. Role of microRNAs in plant and animal development. Science. 2003;301:336–8.

    Article  CAS  PubMed  Google Scholar 

  18. Ma J, Dong C, Ji C. MicroRNA and drug resistance. Cancer Gene Ther. 2010;17:523–31.

    Article  CAS  PubMed  Google Scholar 

  19. Paik WH, Kim HR, Park JK, Song BJ, Lee SH, Hwang JH. Chemosensitivity induced by down-regulation of microRNA-21 in gemcitabine-resistant pancreatic cancer cells by indole-3-carbinol. Anticancer Res. 2013;33:1473–81.

    CAS  PubMed  Google Scholar 

  20. Mittal A, Chitkara D, Behrman SW, Mahato RI. Efficacy of gemcitabine conjugated and miRNA-205 complexed micelles for treatment of advanced pancreatic cancer. Biomaterials. 2014;35:7077–87.

    Article  CAS  PubMed  Google Scholar 

  21. Giovannetti E, Funel N, Peters GJ, Del CM, Erozenci LA, Vasile E, et al. MicroRNA-21 in pancreatic cancer: correlation with clinical outcome and pharmacologic aspects underlying its role in the modulation of gemcitabine activity. Cancer Res. 2010;70:4528–38.

    Article  CAS  PubMed  Google Scholar 

  22. Nagano H, Tomimaru Y, Eguchi H, Hama N, Wada H, Kawamoto K, et al. MicroRNA-29a induces resistance to gemcitabine through the Wnt/beta-catenin signaling pathway in pancreatic cancer cells. Int J Oncol. 2013;43:1066–72.

    CAS  PubMed  Google Scholar 

  23. Li Y, VandenBoom TN, Kong D, Wang Z, Ali S, Philip PA, et al. Up-regulation of miR-200 and let-7 by natural agents leads to the reversal of epithelial-to-mesenchymal transition in gemcitabine-resistant pancreatic cancer cells. Cancer Res. 2009;69:6704–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Ohuchida K, Mizumoto K, Kayashima T, Fujita H, Moriyama T, Ohtsuka T, et al. MicroRNA expression as a predictive marker for gemcitabine response after surgical resection of pancreatic cancer. Ann Surg Oncol. 2011;18:2381–7.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Djuranovic S, Nahvi A, Green R. MiRNA-mediated gene silencing by translational repression followed by mRNA deadenylation and decay. Science. 2012;336:237–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Li Y, Xu J, Chen H, Bai J, Li S, Zhao Z, et al. Comprehensive analysis of the functional microRNA-mRNA regulatory network identifies miRNA signatures associated with glioma malignant progression. Nucleic Acids Res. 2013;41:e203.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Monzo M, Navarro A, Bandres E, Artells R, Moreno I, Gel B, et al. Overlapping expression of microRNAs in human embryonic colon and colorectal cancer. Cell Res. 2008;18:823–33.

    Article  CAS  PubMed  Google Scholar 

  28. Wettenhall JM, Smyth GK. LimmaGUI: a graphical user interface for linear modeling of microarray data. Bioinformatics. 2004;20:3705–6.

    Article  CAS  PubMed  Google Scholar 

  29. Benjamini Y, Drai D, Elmer G, Kafkafi N, Golani I. Controlling the false discovery rate in behavior genetics research. Behav Brain Res. 2001;125:279–84.

    Article  CAS  PubMed  Google Scholar 

  30. Ashburner M, Ball CA, Blake JA, Botstein D, Butler H, Cherry JM, et al. Gene ontology: tool for the unification of biology. The gene ontology consortium. Nat Genet. 2000;25:25–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Draghici S, Khatri P, Tarca AL, Amin K, Done A, Voichita C, et al. A systems biology approach for pathway level analysis. Genome Res. 2007;17:1537–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Kornmann M, Danenberg KD, Arber N, Beger HG, Danenberg PV, Korc M. Inhibition of cyclin D1 expression in human pancreatic cancer cells is associated with increased chemosensitivity and decreased expression of multiple chemoresistance genes. Cancer Res. 1999;59:3505–11.

    CAS  PubMed  Google Scholar 

  33. Jimeno A, Hidalgo M. Molecular biomarkers: their increasing role in the diagnosis, characterization, and therapy guidance in pancreatic cancer. Mol Cancer Ther. 2006;5:787–96.

    Article  CAS  PubMed  Google Scholar 

  34. Brody JR, Witkiewicz AK, Yeo CJ. The past, present, and future of biomarkers: a need for molecular beacons for the clinical management of pancreatic cancer. Adv Surg. 2011;45:301–21.

    Article  PubMed  Google Scholar 

  35. Dong Y, Stephens C, Walpole C, Swedberg JE, Boyle GM, Parsons PG, et al. Paclitaxel resistance and multicellular spheroid formation are induced by kallikrein-related peptidase 4 in serous ovarian cancer cells in an ascites mimicking microenvironment. PLoS ONE. 2013;8:e57056.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Ho AS, Huang X, Cao H, Christman-Skieller C, Bennewith K, Le QT, et al. Circulating miR-210 as a novel hypoxia marker in pancreatic cancer. Transl Oncol. 2010;3:109–13.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Boren T, Xiong Y, Hakam A, Wenham R, Apte S, Chan G, et al. MicroRNAs and their target messenger RNAs associated with ovarian cancer response to chemotherapy. Gynecol Oncol. 2009;113:249–55.

    Article  CAS  PubMed  Google Scholar 

  38. Xia L, Zhang D, Du R, Pan Y, Zhao L, Sun S, et al. MiR-15b and miR-16 modulate multidrug resistance by targeting BCL2 in human gastric cancer cells. Int J Cancer. 2008;123:372–9.

    Article  CAS  PubMed  Google Scholar 

  39. Wang H, Zhu LJ, Yang YC, Wang ZX, Wang R. MiR-224 promotes the chemoresistance of human lung adenocarcinoma cells to cisplatin via regulating G(1)/S transition and apoptosis by targeting p21(WAF1/CIP1). Br J Cancer. 2014;111:339–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Hong L, Yang Z, Ma J, Fan D. Function of miRNA in controlling drug resistance of human cancers. Curr Drug Targets. 2013;14:1118–27.

    Article  CAS  PubMed  Google Scholar 

  41. Tang D, Kang R, Livesey KM, Zeh HR, Lotze MT. High mobility group box 1 (HMGB1) activates an autophagic response to oxidative stress. Antioxid Redox Signal. 2011;15:2185–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Kang CB, Feng L, Chia J, Yoon HS. Molecular characterization of FK-506 binding protein 38 and its potential regulatory role on the anti-apoptotic protein Bcl-2. Biochem Biophys Res Commun. 2005;337:30–8.

    Article  CAS  PubMed  Google Scholar 

  43. Shi C, Ma Y, Liu H, Zhang Y, Wang Z, Jia H. The non-receptor tyrosine kinase c-Src mediates the PDGF-induced association between Furin and pro-MT1-MMP in HPAC pancreatic cells. Mol Cell Biochem. 2012;362:65–70.

    Article  CAS  PubMed  Google Scholar 

  44. Zhang YT, Xu LH, Lu Q, Liu KP, Liu PY, Ji F, et al. VASP activation via the Galpha13/RhoA/PKA pathway mediates cucurbitacin-B-induced actin aggregation and cofilin-actin rod formation. PLoS ONE. 2014;9:e93547.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This study was supported by the National Science Fundation of China grant 81102685 (PI Yehua Shen).

Conflicts of interest

The authors have declared no conflicts of interest.

Author information

Authors and Affiliations

  1. Department of Integrative Oncology, Fudan University Shanghai Cancer Center, Shanghai, 200032, China

    Yehua Shen, Yan Pan, Litao Xu, Lianyu Chen, Luming Liu, Hao Chen, Zhen Chen & Zhiqiang Meng

  2. Department of Oncology, Shanghai Medical College, Fudan University, 270 Dong’An Road, Shanghai, 200032, China

    Yehua Shen, Yan Pan, Litao Xu, Lianyu Chen, Luming Liu, Hao Chen, Zhen Chen & Zhiqiang Meng

Authors
  1. Yehua Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yan Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Litao Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lianyu Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Luming Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hao Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Zhen Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Zhiqiang Meng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhiqiang Meng.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(XLS 259 kb)

ESM 2

(XLS 34 kb)

ESM 3

(XLS 209 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shen, Y., Pan, Y., Xu, L. et al. Identifying microRNA-mRNA regulatory network in gemcitabine-resistant cells derived from human pancreatic cancer cells. Tumor Biol. 36, 4525–4534 (2015). https://doi.org/10.1007/s13277-015-3097-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13277-015-3097-8

Keywords

Search

Navigation