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World Journal of Surgery

, 33:698 | Cite as

Profiling of 95 MicroRNAs in Pancreatic Cancer Cell Lines and Surgical Specimens by Real-Time PCR Analysis

  • Yuqing Zhang
  • Min Li
  • Hao Wang
  • William E. Fisher
  • Peter H. Lin
  • Qizhi Yao
  • Changyi ChenEmail author
Article

Abstract

Background

MicroRNAs (miRNAs) are involved in cancer pathogenesis, apoptosis, and cell growth, thereby functioning as tumor suppressors or oncogenes. However, expression alterations and roles of these miRNAs in pancreatic cancer are largely unknown. We hypothesized that pancreatic cancer may have a unique miRNA profile, which may play a critical role in pancreatic cancer development, progression, diagnosis, and prognosis.

Methods

Differential expression of 95 miRNAs was analyzed by real time RT-PCR using the QuantiMir System. All 95 miRNAs chosen for the array are based on their potential functions related to cancer biology, cell development, and apoptosis. The expression of miRNAs for pancreatic cancer tissue samples or cancer cell lines was normalized to U6 RNA and compared with those in relatively normal pancreatic tissues or normal human pancreatic ductal epithelial (HPDE) cells. Human pancreatic tissue with chronic pancreatitis also was included for analysis.

Results

In the initial analysis, the expression of most 95 miRNAs was substantially changed in pancreatic cancer tissues (n = 5) and cell lines (n = 3) compared with relatively normal pancreatic tissues and HPDE cells. However, each pancreatic cancer tissue or cell type had a substantially different profiling pattern with other cases or cell types as well as chronic pancreatitis tissue, indicating the individual diversity of pancreatic cancer. Further analysis was performed on 10 pancreatic cancer cell lines and 17 pairs of pancreatic cancer/normal tissues. Eight miRNAs were significantly upregulated in most pancreatic cancer tissues and cell lines, including miR-196a, miR-190, miR-186, miR-221, miR-222, miR-200b, miR-15b, and miR-95. The incidence of upregulation of these eight genes between normal control subjects and tumor cells or tissues ranged from 70–100%. The magnitude of increase of these miRNAs in pancreatic cancer samples ranged from 3- to 2018-fold of normal control subjects.

Conclusions

Pancreatic cancer tissues or cell lines have a unique miRNA profiling pattern at the individual basis compared with relatively normal pancreatic tissues or cells as well as pancreatitis tissue. Upregulation of eight miRNAs occurs in most pancreatic cancer tissues and cell types. These miRNAs may share common pathways in pancreatic cancer pathogenesis. This study may provide useful information for further investigations of functional roles of miRNAs in pancreatic cancer development, progression, diagnosis, and prognosis.

Keywords

Pancreatic Cancer Chronic Pancreatitis miRNA Expression Pancreatic Cancer Cell Line Pancreatic Cancer Tissue 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This study was partially supported by the Michael E. DeBakey Department of Surgery at the Baylor College of Medicine and the Michael E. DeBakey VA Medical Center, Houston, Texas, USA.

References

  1. 1.
    Esquela-Kerscher A, Slack FJ (2006) Oncomirs: microRNAs with a role in cancer. Nat Rev Cancer 6:259–269PubMedCrossRefGoogle Scholar
  2. 2.
    Sevignani C, Calin GA, Siracusa LD, Croce CM (2006) Mammalian microRNAs: a small world for fine-tuning gene expression. Mamm Genome 17:189–202PubMedCrossRefGoogle Scholar
  3. 3.
    Barbarotto E, Schmittgen TD, Calin GA (2008) MicroRNAs and cancer: profile, profile, profile. Int J Cancer 122:969–977PubMedCrossRefGoogle Scholar
  4. 4.
    Kent OA, Mendell JT (2006) A small piece in the cancer puzzle: microRNAs as tumor suppressors and oncogenes. Oncogene 25:6188–6196PubMedCrossRefGoogle Scholar
  5. 5.
    Valencia-Sanchez MA, Liu J, Hannon GJ, Parker R (2006) Control of translation and mRNA degradation by miRNAs and siRNAs. Genes Dev 20:515–524PubMedCrossRefGoogle Scholar
  6. 6.
    Hornstein E, Mansfield JH, Yekta S et al (2005) The microRNA miR-196 acts upstream of Hoxb8 and Shh in limb development. Nature 438:671–674PubMedCrossRefGoogle Scholar
  7. 7.
    Datta J, Kutay H, Nasser MW et al (2008) Methylation mediated silencing of MicroRNA-1 gene and its role in hepatocellular carcinogenesis. Cancer Res 68:5049–5058PubMedCrossRefGoogle Scholar
  8. 8.
    Calin GA, Ferracin M, Cimmino A et al (2005) A MicroRNA signature associated with prognosis and progression in chronic lymphocytic leukemia. N Engl J Med 353:1793–1801PubMedCrossRefGoogle Scholar
  9. 9.
    Jeffrey SS (2008) Cancer biomarker profiling with microRNAs. Nat Biotechnol 26:400–401PubMedCrossRefGoogle Scholar
  10. 10.
    Ma L, Teruya-Feldstein J, Weinberg RA (2007) Tumour invasion and metastasis initiated by microRNA-10b in breast cancer. Nature 449:682–688PubMedCrossRefGoogle Scholar
  11. 11.
    Lu J, Getz G, Miska EA et al (2005) MicroRNA expression profiles classify human cancers. Nature 435:834–838PubMedCrossRefGoogle Scholar
  12. 12.
    Saif MW (2006) Pancreatic cancer: highlights from the 42nd annual meeting of the American Society of Clinical Oncology, 2006. JOP 7:337–348PubMedGoogle Scholar
  13. 13.
    Jemal A, Tiwari RC, Murray T et al (2004) Cancer statistics, 2004. CA Cancer J Clin 54:8–29PubMedCrossRefGoogle Scholar
  14. 14.
    Sun M, Estrov Z, Ji Y et al (2008) Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells. Mol Cancer Ther 7:464–473PubMedCrossRefGoogle Scholar
  15. 15.
    Gironella M, Seux M, Xie MJ et al (2007) Tumor protein 53-induced nuclear protein 1 expression is repressed by miR-155, and its restoration inhibits pancreatic tumor development. Proc Natl Acad Sci USA 104:16170–16175PubMedCrossRefGoogle Scholar
  16. 16.
    Bloomston M, Frankel WL, Petrocca F et al (2007) MicroRNA expression patterns to differentiate pancreatic adenocarcinoma from normal pancreas and chronic pancreatitis. JAMA 297:1901–1908PubMedCrossRefGoogle Scholar
  17. 17.
    Szafranska AE, Davison TS, John J et al (2007) MicroRNA expression alterations are linked to tumorigenesis and non-neoplastic processes in pancreatic ductal adenocarcinoma. Oncogene 26:4442–4452PubMedCrossRefGoogle Scholar
  18. 18.
    Lee EJ, Gusev Y, Jiang J et al (2007) Expression profiling identifies microRNA signature in pancreatic cancer. Int J Cancer 120:1046–1054PubMedCrossRefGoogle Scholar
  19. 19.
    Tsuda N, Ishiyama S, Li Y et al (2006) Synthetic microRNA designed to target glioma-associated antigen 1 transcription factor inhibits division and induces late apoptosis in pancreatic tumor cells. Clin Cancer Res 12:6557–6564PubMedCrossRefGoogle Scholar
  20. 20.
    Roldo C, Missiaglia E, Hagan JP et al (2006) MicroRNA expression abnormalities in pancreatic endocrine and acinar tumors are associated with distinctive pathologic features and clinical behavior. J Clin Oncol 24:4677–4684PubMedCrossRefGoogle Scholar
  21. 21.
    Furukawa T, Duguid WP, Rosenberg L et al (1996) Long-term culture and immortalization of epithelial cells from normal adult human pancreatic ducts transfected by the E6E7 gene of human papillomavirus 16. Am J Pathol 148:1763–1770PubMedGoogle Scholar
  22. 22.
    Ouyang H, Mou L, Luk C et al (2000) Immortal human pancreatic duct epithelial cell lines with near normal genotype and phenotype. Am J Pathol 157:1623–3123PubMedGoogle Scholar
  23. 23.
    Li M, Bharadwaj U, Zhang R et al (2008) Mesothelin is a malignant factor and therapeutic vaccine target for pancreatic cancer. Mol Cancer Ther 7:286–296PubMedCrossRefGoogle Scholar
  24. 24.
    Li M, Zhang Y, Liu Z et al (2007) Aberrant expression of zinc transporter ZIP4 (SLC39A4) significantly contributes to human pancreatic cancer pathogenesis and progression. Proc Natl Acad Sci USA 104:18636–18641PubMedCrossRefGoogle Scholar
  25. 25.
    Li M, Yang H, Chai H et al (2004) Pancreatic carcinoma cells express neuropilins and vascular endothelial growth factor, but not vascular endothelial growth factor receptors. Cancer 101:2341–2350PubMedCrossRefGoogle Scholar
  26. 26.
    Li M, Zhai Q, Bharadwaj U et al (2006) Cyclophilin A is overexpressed in human pancreatic cancer cells and stimulates cell proliferation through CD147. Cancer 106:2284–2294PubMedCrossRefGoogle Scholar
  27. 27.
    Hobert O (2004) Common logic of transcription factor and microRNA action. Trends Biochem Sci 29:462–468PubMedCrossRefGoogle Scholar
  28. 28.
    Mansfield JH, Harfe BD, Nissen R et al (2004) MicroRNA-responsive “sensor” transgenes uncover Hox-like and other developmentally regulated patterns of vertebrate microRNA expression. Nat Genet 36:1079–1083PubMedCrossRefGoogle Scholar
  29. 29.
    Gregory PA, Bert AG, Paterson EL et al (2008) The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol 10:593–601PubMedCrossRefGoogle Scholar
  30. 30.
    Xia L, Zhang D, Du R et al (2008) miR-15b and miR-16 modulate multidrug resistance by targeting BCL2 in human gastric cancer cells. Int J Cancer 123:372–379PubMedCrossRefGoogle Scholar
  31. 31.
    Cheng AM, Byrom MW, Shelton J, Ford LP (2005) Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis. Nucleic Acids Res 33:1290–1297PubMedCrossRefGoogle Scholar
  32. 32.
    Ciafre SA, Galardi S, Mangiola A et al (2005) Extensive modulation of a set of microRNAs in primary glioblastoma. Biochem Biophys Res Commun 334:1351–1358PubMedCrossRefGoogle Scholar
  33. 33.
    He H, Jazdzewski K, Li W et al (2005) The role of microRNA genes in papillary thyroid carcinoma. Proc Natl Acad Sci USA 102:19075–19080PubMedCrossRefGoogle Scholar
  34. 34.
    Krutzfeldt J, Rajewsky N, Braich R et al (2005) Silencing of microRNAs in vivo with “antagomirs”. Nature 438:685–689PubMedCrossRefGoogle Scholar

Copyright information

© Société Internationale de Chirurgie 2008

Authors and Affiliations

  • Yuqing Zhang
    • 1
  • Min Li
    • 1
  • Hao Wang
    • 1
  • William E. Fisher
    • 1
  • Peter H. Lin
    • 1
  • Qizhi Yao
    • 1
  • Changyi Chen
    • 1
    Email author
  1. 1.Michael E. DeBakey Department of Surgery, Division of Vascular Surgery and Endovascular Therapy, Molecular Surgeon Research CenterBaylor College of Medicine and Michael E. DeBakey VA Medical CenterHoustonUSA

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