MiR-21 Upregulation Induced by Promoter Zone Histone Acetylation is Associated with Chemoresistance to Gemcitabine and Enhanced Malignancy of Pancreatic Cancer Cells

  • Song, Wei-Feng (Department of Medical Oncology, Shanghai First People's Hospital, Shanghai Jiao Tong University) ;
  • Wang, Lei (Department of Medical Oncology, Shanghai First People's Hospital, Shanghai Jiao Tong University) ;
  • Huang, Wei-Yi (Department of Medical Oncology, Shanghai First People's Hospital, Shanghai Jiao Tong University) ;
  • Cai, Xun (Department of Medical Oncology, Shanghai First People's Hospital, Shanghai Jiao Tong University) ;
  • Cui, Jiu-Jie (Department of Medical Oncology, Shanghai First People's Hospital, Shanghai Jiao Tong University) ;
  • Wang, Li-Wei (Department of Medical Oncology, Shanghai First People's Hospital, Shanghai Jiao Tong University)
  • Published : 2013.12.31


Background and Aims: MicroRNA-21 (miR-21) is reported to be overexpressed and to contribute to proliferation, apoptosis and gemcitabine resistance in pancreatic ductal adenocarcinomas (PDACs). The aims of this study were to explore regulation of miR-21 expression by epigenetic change and its impact on chemoresistance and malignant properties of of pancreatic cancer. Materials and methods: We retrospectively collected 41 cases of advanced pancreatic cancer patients who were sensitive or resistant to gemcitabine and assessed levels of serum circulating miR-21 for correlation with cytotoxic activity. Histone acetylation in the miR-21 promoter was also studied in gemcitabine-sensitive and gemcitabine-resistant PDAC cells. Gemcitabine-resistant HPAC and PANC-1 cells were transfected with pre-miR-21 precursors (pre-miR-21) and antisense oligonucleotides (anti-miR-21), and were treated with TSA. Finally, invasion and metastasis assays were performed and alteration in mir-21, PTEN, AKT and pAKT level was evaluated in these cells. Results: Serum miR-21 levels were increased in gemcitabine-resistant PDAC patients compared with gemcitabine-sensitive subjects. The miR-21 levels were increased in 6 PDAC cells treated with gemcitabine significantly, associated with 50% inhibitory concentrations ($IC_{50}s$). Histone acetylation levels at miR-21 promoter were increased in PDAC cells after treatment with gemcitabine. Enhanced invasion and metastasis, increased miR-21 expression, decreased PTEN, elevated pAKT level were demonstrated in gemcitabine-resistant HPAC and PANC-1 cells. Pre-miR-21 transfection or TSA treatment further increased invasion and metastasis ability, decreased PTEN, and elevated pAKT levels in these two lines. In contrast, anti-miR-21 transfection could reverse invasion and metastasis, and PTEN and pAKT expressions induced by gemcitabine. Conclusions: MiR-21 upregulation induced by histone acetylation in the promoter zone is associated with chemoresistance to gemcitabine and enhanced malignant potential in pancreatic cancer cells.


  1. Bader AG, Kang S, Zhao L, Vogt PK (2005). Oncogenic PI3K deregulates transcription and translation. Nature reviews. Cancer, 5, 921-9.
  2. Bartel DP (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Cell, 116, 281-97.
  3. Ali S, Ahmad A, Banerjee S, et al (2010). Gemcitabine sensitivity can be induced in pancreatic cancer cells through modulation of miR-200 and miR-21 expression by curcumin or its analogue CDF. Cancer Res, 70, 3606-17.
  4. Asangani IA, Rasheed SA, Nikolova DA, et al (2008). MicroRNA-21 (miR-21) post-transcriptionally downregulates tumor suppressor Pdcd4 and stimulates invasion, intravasation and metastasis in colorectal cancer. Oncogene, 27, 2128-36.
  5. Bloomston M, Frankel WL, Petrocca F, et al (2007). MicroRNA expression patterns to differentiate pancreatic adenocarcinoma from normal pancreas and chronic pancreatitis. JAMA:J Am Med Assoc, 297, 1901-8.
  6. Boumber Y, Issa JP (2011). Epigenetics in cancer: what's the future? Oncology (Williston Park, N.Y.), 25, 220-6, 228.
  7. Burris HA, 3rd Moore MJ, Andersen J, et al (1997). Improvements in survival and clinical benefit with gemcitabine as first-line therapy for patients with advanced pancreas cancer: a randomized trial. J Clin Oncol, 15, 2403-13.
  8. Chakrabarti SK, James JC, Mirmira RG (2002). Quantitative assessment of gene targeting in vitro and in vivo by the pancreatic transcription factor, Pdx1. Importance of chromatin structure in directing promoter binding. J Biol Chem, 277, 13286-93.
  9. Chan SH, Wu CW, Li AF, Chi CW, Lin WC (2008). miR-21 microRNA expression in human gastric carcinomas and its clinical association. Anticancer Res, 28, 907-11.
  10. Chen X, Ba Y, Ma L, et al (2008). Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res, 18, 997-1006.
  11. Cho WC (2010). MicroRNAs in cancer - from research to therapy. BBA, 1805, 209-17.
  12. Tomimaru Y, Eguchi H, Nagano H, et al (2010). MicroRNA-21 induces resistance to the anti-tumour effect of interferonalpha/5-fluorouracil in hepatocellular carcinoma cells. Br J Cancer, 103, 1617-26.
  13. Vogt PK, Gymnopoulos M, Hart JR (2009). PI 3-kinase and cancer: changing accents. Curr Opin Genet Dev, 19, 12-7.
  14. Wang P, Zhuang L, Zhang J, et al (2013a). The serum miR-21 level serves as a predictor for the chemosensitivity of advanced pancreatic cancer, and miR-21 expression confers chemoresistance by targeting FasL. Mol Oncol, 7, 334-45.
  15. Wang Z, Yao H, Lin S, et al (2013b). Transcriptional and epigenetic regulation of human microRNAs. Cancer Lett, 331, 1-10.
  16. Wickramasinghe NS, Manavalan TT, Dougherty SM, et al (2009). Estradiol downregulates miR-21 expression and increases miR-21 target gene expression in MCF-7 breast cancer cells. Nucleic Acids Res, 37, 2584-95.
  17. Yang CH, Yue J, Pfeffer SR, Handorf CR, Pfeffer LM (2011). MicroRNA miR-21 regulates the metastatic behavior of B16 melanoma cells. J Biol Chem, 286, 39172-8.
  18. Yoo CB, Jones PA (2006). Epigenetic therapy of cancer: past, present and future. Nature Rev Drug Discov, 5, 37-50.
  19. Zhang BG, Li JF, Yu BQ, et al (2012). microRNA-21 promotes tumor proliferation and invasion in gastric cancer by targeting PTEN. Oncology Rep, 27, 1019-26.
  20. Zhang XH, Rao M, Loprieato JA, et al (2008a). Aurora A, Aurora B and survivin are novel targets of transcriptional regulation by histone deacetylase inhibitors in non-small cell lung cancer. Cancer Biol Ther, 7, 1388-97.
  21. Zhang Z, Li Z, Gao C, et al (2008b). miR-21 plays a pivotal role in gastric cancer pathogenesis and progression. Lab Invest; a journal of technical methods and pathology, 88, 1358-66.
  22. Zhu S, Si ML, Wu H, Mo YY (2007). MicroRNA-21 targets the tumor suppressor gene tropomyosin 1 (TPM1). J Biol Chem, 282, 14328-36.
  23. Zhu S, Wu H, Wu F, et al (2008). MicroRNA-21 targets tumor suppressor genes in invasion and metastasis. Cell Res, 18, 350-9.
  24. Lim LP, Lau NC, Garrett-Engele P, et al (2005). Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature, 433, 769-73.
  25. Liu ZL, Wang H, Liu J, Wang ZX (2013). MicroRNA-21 (miR-21). expression promotes growth, metastasis, and chemoor radioresistance in non-small cell lung cancer cells by targeting PTEN. Mol Cell Biochem, 372, 35-45.
  26. Mardin WA, Mees ST (2009). MicroRNAs: novel diagnostic and therapeutic tools for pancreatic ductal adenocarcinoma? Ann Surg Oncol, 16, 3183-9.
  27. Meng F, Henson R, Lang M, et al (2006). Involvement of human micro-RNA in growth and response to chemotherapy in human cholangiocarcinoma cell lines. Gastroenterology, 130, 2113-29.
  28. Meng F, Henson R, Wehbe-Janek H, et al (2007). MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology, 133, 647-58.
  29. Mitchell PS, Parkin RK, Kroh EM, et al (2008). Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci USA, 105, 10513-8.
  30. Miyanaga A, Gemma A, Noro R, et al (2008). Antitumor activity of histone deacetylase inhibitors in non-small cell lung cancer cells: development of a molecular predictive model. Mol Cancer Ther, 7, 1923-30.
  31. Moriyama T, Ohuchida K, Mizumoto K, et al (2009). MicroRNA-21 modulates biological functions of pancreatic cancer cells including their proliferation, invasion, and chemoresistance. Mol Cancer Ther, 8, 1067-74.
  32. Nagao Y, Hisaoka M, Matsuyama A, et al (2012). Association of microRNA-21 expression with its targets, PDCD4 and TIMP3, in pancreatic ductal adenocarcinoma. Modern Pathol, 25, 112-21.
  33. Negrini M, Ferracin M, Sabbioni S, Croce CM (2007). MicroRNAs in human cancer: from research to therapy. J Cell Sci, 120, 1833-40.
  34. Ng SS, Tsao MS, Nicklee T, Hedley DW (2001). Wortmannin inhibits pkb/akt phosphorylation and promotes gemcitabine antitumor activity in orthotopic human pancreatic cancer xenografts in immunodeficient mice. Clin Cancer Res, 7, 3269-75.
  35. Siegel R, Naishadham D, Jemal A (2012). Cancer statistics, 2012. CA Cancer J Clin, 62, 10-29.
  36. Slaby O, Svoboda M, Fabian P, et al (2007). Altered expression of miR-21, miR-31, miR-143 and miR-145 is related to clinicopathologic features of colorectal cancer. Oncology, 72, 397-402.
  37. Szafranska AE, Davison TS, John J, et al (2007). MicroRNA expression alterations are linked to tumorigenesis and nonneoplastic processes in pancreatic ductal adenocarcinoma. Oncogene, 26, 4442-52.
  38. Li J, Huang H, Sun L, et al (2009). MiR-21 indicates poor prognosis in tongue squamous cell carcinomas as an apoptosis inhibitor. Clin Cancer Res, 15, 3998-4008.
  39. Li L, Ernsting BR, Wishart MJ, Lohse DL, Dixon JE (1997). A family of putative tumor suppressors is structurally and functionally conserved in humans and yeast. J Biol Chem, 272, 29403-6.
  40. Choi YH (2005). Induction of apoptosis by trichostatin A, a histone deacetylase inhibitor, is associated with inhibition of cyclooxygenase-2 activity in human non-small cell lung cancer cells. Int J Oncol, 27, 473-9.
  41. Dillhoff M, Liu J, Frankel W, Croce C, Bloomston M (2008). MicroRNA-21 is overexpressed in pancreatic cancer and a potential predictor of survival. J Gastrointest Surg, 12, 2171-6.
  42. Funel N, Giovannetti E, Del Chiaro M, et al (2008). Laser microdissection and primary cell cultures improve pharmacogenetic analysis in pancreatic adenocarcinoma. Lab Invest, 88, 773-84.
  43. Gabriely G, Wurdinger T, Kesari S, et al (2008). MicroRNA 21 promotes glioma invasion by targeting matrix metalloproteinase regulators. Mol Cell Biol, 28, 5369-80.
  44. Giovannetti E, Del Tacca M, Mey V, et al (2006). Transcription analysis of human equilibrative nucleoside transporter-1 predicts survival in pancreas cancer patients treated with gemcitabine. Cancer Res, 66, 3928-35.
  45. Giovannetti E, Funel N, Peters GJ, et al (2010). MicroRNA-21 in pancreatic cancer: correlation with clinical outcome and pharmacologic aspects underlying its role in the modulation of gemcitabine activity. Cancer Res, 70, 4528-38.
  46. Hiyoshi Y, Kamohara H, Karashima R, et al (2009). MicroRNA-21 regulates the proliferation and invasion in esophageal squamous cell carcinoma. Clin Cancer Res, 15, 1915-22.
  47. Hwang JH, Voortman J, Giovannetti E, et al (2010). Identification of microRNA-21 as a biomarker for chemoresistance and clinical outcome following adjuvant therapy in resectable pancreatic cancer. PloS one, 5, e10630.
  48. Iorio MV, Visone R, Di Leva G, et al (2007). MicroRNA signatures in human ovarian cancer. Cancer Res, 67, 8699-707.
  49. Isozaki Y, Hoshino I, Nohata N, et al (2012). Identification of novel molecular targets regulated by tumor suppressive miR-375 induced by histone acetylation in esophageal squamous cell carcinoma. Int J Oncol, 41, 985-94.
  50. Jamieson NB, Morran DC, Morton JP, et al (2012). MicroRNA molecular profiles associated with diagnosis, clinicopathologic criteria, and overall survival in patients with resectable pancreatic ductal adenocarcinoma. Clin Cancer Res, 18, 534-45.
  51. Krichevsky AM, Gabriely G (2009). miR-21: a small multifaceted RNA. J Cell Mol Med, 13, 39-53.
  52. Kwak HJ, Kim YJ, Chun KR, et al (2011). Downregulation of Spry2 by miR-21 triggers malignancy in human gliomas. Oncogene, 30, 2433-42.
  53. Lane AA, Chabner BA (2009). Histone deacetylase inhibitors in cancer therapy. J Clin Oncol, 27, 5459-68.
  54. Leslie NR, Downes CP (2002). PTEN: The down side of PI 3-kinase signalling. Cell Signal, 14, 285-95.
  55. Li D, Xie K, Wolff R, Abbruzzese JL (2004). Pancreatic cancer. Lancet, 363, 1049-57.

Cited by

  1. Associations between Single Nucleotide Polymorphisms of COX-2 and MMP-2 Genes and Colorectal Cancer Susceptibility in the Saudi Population vol.15, pp.12, 2014,
  2. MicroRNAs as Promising Biomarkers for Tumor-staging: Evaluation of MiR21 MiR155 MiR29a and MiR92a in Predicting Tumor Stage of Rectal Cancer vol.15, pp.13, 2014,
  3. Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling vol.15, pp.18, 2014,
  4. Induction of MicroRNA-9 Mediates Cytotoxicity of Curcumin Against SKOV3 Ovarian Cancer Cells vol.15, pp.8, 2014,
  5. miR-21 Is Overexpressed in Hydatidiform Mole Tissues and Promotes Proliferation, Migration, and Invasion in Choriocarcinoma Cells vol.27, pp.2, 2017,
  6. Gemcitabine Enhances Kras-MEK–Induced Matrix Metalloproteinase-10 Expression Via Histone Acetylation in Gemcitabine-Resistant Pancreatic Tumor-initiating Cells vol.46, pp.2, 2017,
  7. Critical role of HMGA proteins in cancer cell chemoresistance vol.95, pp.4, 2017,
  8. miRNA-21 enhances chemoresistance to cisplatin in epithelial ovarian cancer by negatively regulating PTEN vol.14, pp.2, 2017,
  9. The role and mechanisms of action of microRNAs in cancer drug resistance vol.11, pp.1, 2019,