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Medical Oncology

, Volume 29, Issue 5, pp 3113–3118 | Cite as

The quantitative analysis by stem-loop real-time PCR revealed the microRNA-34a, microRNA-155 and microRNA-200c overexpression in human colorectal cancer

  • Mojin Wang
  • Peng Zhang
  • Yuan Li
  • Guanghui Liu
  • Bin Zhou
  • Lan Zhan
  • Zongguang Zhou
  • Xiaofeng Sun
Original Paper

Abstract

The recently identified class of microRNAs (miRNAs) provided a new insight in cancer research. As the member of miRNAs family, miR-34a, miR-155 and miR-200c abnormalities have been found in various types of cancer. However, the relationship between these three miRNAs (miR-34a, miR-155 and miR-200c) and colorectal cancer is unclear. In this study, we applied stem-loop real-time PCR to quantitatively detect miR-34a, miR-155 and miR-200c expression in 109 pair-matched human colorectal cancers and the corresponding normal mucosa. MiR-34a (2.2-fold), miR-155 (2.3-fold) and miR-200c (3.1-fold) were all expressed at higher levels in colorectal cancer (P = 0.001, 0.005 and 0.001, respectively). In rectum, miR-34a and miR-200c were significantly upregulated (P = 0.006 and 0.007), while the miR-155 overexpression was not statistically significant (P = 0.083). In colon, the higher expression of three miRNAs was seen, however, without significant difference (P > 0.05). We also found that the miR-34a expression was higher in rectal cancer having more advanced TNM stage (III + IV, P = 0.03). Then miR-200c expression was positively correlated with and sera CEA level of rectal cancer patients (P = 0.04). In conclusion, our results thus suggest that the overexpression of miR-34a, miR-155 and miR-200c be associated with the development of colorectal cancer, meanwhile miR-34a may be involved in the development and progression of rectal cancer. The more deeply and larger scale research are required to prove the correlation.

Keywords

MicroRNA-34a MicroRNA-155 MicroRNA-200c Colorectal cancer Stem-loop real-time RT-PCR 

Notes

Acknowledgments

Grant sponsor: National Natural Science Foundation of China (No. 30830100); Ph.D. Programs Foundation of Ministry of Education of China (No. 200806100058).

Conflict of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

Supplementary material

12032_2012_241_MOESM1_ESM.tif (563 kb)
Supplementary Fig. The sequence of miR-34a (a), miR-155 (b) and miR-200c (c) stem-loop RT primers (TIFF 563 kb)
12032_2012_241_MOESM2_ESM.doc (36 kb)
Supplementary material 2 (DOC 35 kb)

References

  1. 1.
    Novina CD, Sharp PA. The RNAi revolution. Nature. 2004;430:161–4.PubMedCrossRefGoogle Scholar
  2. 2.
    Yekta S, Shih I, Bartel DP. MicroRNA-directed cleavage of HOXB8 mRNA. Science. 2004;304:594.PubMedCrossRefGoogle Scholar
  3. 3.
    Cheng AM, Byrom MW, Shelton J, Ford LP. Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis. Nucleic Acids Res. 2005;33:1290–7.PubMedCrossRefGoogle Scholar
  4. 4.
    Chen CZ, Li L, Lodish HF, Bartel DP. MicroRNAs modulate hematopoietic lineage differentiation. Science. 2004;303:83–6.PubMedCrossRefGoogle Scholar
  5. 5.
    Bandres E, Agirre X, Bitarte N, Ramirez N, Zarate R, Roman-Gomez J, et al. Epigenetic regulation of microRNA expression in colorectal cancer. Int J Cancer. 2009;125:2737–43.PubMedCrossRefGoogle Scholar
  6. 6.
    Dostie J, Mourelatos Z, Yang M, Sharma A, Dreyfuss G. Numerous microRNPs in neuronal cells containing novel microRNAs. RNA. 2003;9:180–6.PubMedCrossRefGoogle Scholar
  7. 7.
    Lagos-Quintana M, Rauhut R, Yalcin A, Meyer J, Lendeckel W, Tuschl T. Identification of tissue-specific microRNAs from mouse. Curr Biol. 2002;12:735–9.PubMedCrossRefGoogle Scholar
  8. 8.
    Tazawa H, Tsuchiya N, Izumiya M, Nakagama H. Tumorsuppressive miR-34a induces senescence-like growth arrest through modulation of the E2F pathway in human colon cancer cells. PNAS. 2007;104:15472–7.PubMedCrossRefGoogle Scholar
  9. 9.
    Schetter AJ, Leung SY, Sohn JJ, Zanetti KA, Bowman ED, Yanaihara N, et al. MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma. JAMA. 2008;299:425–36.PubMedCrossRefGoogle Scholar
  10. 10.
    Faraoni I, Antonetti FR, Cardone J, Bonmassar E. miR-155 gene: a typical multifunctional microRNA. Biochim Biophys Acta. 2009;1792:497–505.PubMedCrossRefGoogle Scholar
  11. 11.
    Volinia S, Calin GA, Liu CG, Ambs S, Cimmino A, Petrocca F, et al. A microRNA expression signature of human solid tumors defines cancer gene targets. PNAS. 2006;103:2257–61.PubMedCrossRefGoogle Scholar
  12. 12.
    Hu X, Macdonald DM, Huettner PC, Feng Z, El Naqa IM, Schwarz JK, et al. A miR-200 microRNA cluster as prognostic marker in advanced ovarian cancer. Gynecol Oncol. 2009;11:457–64.CrossRefGoogle Scholar
  13. 13.
    Nakajima G, Hayashi K, Xi Y, Kudo K, Uchida K, Takasaki K, et al. Non-coding MicroRNAs hsa-let-7 g and hsa-miR-181b are associated with chemoresponse to S-1 in colon cancer. Cancer Genomics Proteomics. 2006;3:317–24.PubMedGoogle Scholar
  14. 14.
    Xi Y, Formentini A, Chien M, Weir DB, Russo JJ, Ju J, et al. Prognostic values of microRNAs in colorectal cancer. Biomark Insights. 2006;2:113–21.PubMedGoogle Scholar
  15. 15.
    Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, et al. Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 2005;33:e179.PubMedCrossRefGoogle Scholar
  16. 16.
    Compton CC, Greene FL. The staging of colorectal cancer: 2004 and beyond. CA Cancer J Clin. 2004;54:295–308.PubMedCrossRefGoogle Scholar
  17. 17.
    Wang CJ, Zhou ZG, Wang L, Yang L, Zhou B, Gu J, et al. Clinicopathological significance of microRNA-31, -143 and -145 expression in colorectal cancer. Dis Markers. 2009;26:27–34.PubMedGoogle Scholar
  18. 18.
    Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res. 2002;30:e36.PubMedCrossRefGoogle Scholar
  19. 19.
    Yang N, Coukos G, Zhang L. MicroRNA epigenetic alterations in human cancer: one step forward in diagnosis and treatment. Int J Cancer. 2008;122:963–8.PubMedCrossRefGoogle Scholar
  20. 20.
    Yang L, Belaguli N, Berger DH. MicroRNA and colorectal cancer. World J Surg. 2009;33:638–46.PubMedCrossRefGoogle Scholar
  21. 21.
    Davidson LA, Wang N, Shah MS, Lupton JR, Ivanov I, Chapkin RS. n-3 Polyunsaturated fatty acids modulate carcinogen-directed non-coding microRNA signatures in rat colon. Carcinogenesis. 2009;30:2077–84.PubMedCrossRefGoogle Scholar
  22. 22.
    Dutta KK, Zhong Y, Liu YT, Yamada T, Akatsuka S, Hu Q, et al. Association of microRNA-34a overexpression with proliferation is cell type-dependent. Cancer Sci. 2007;98:1845–52.PubMedCrossRefGoogle Scholar
  23. 23.
    Raver-Shapira N, Marciano E, Meiri E, Spector Y, Rosenfeld N, Moskovits N, et al. Transcriptional activation of miR-34a contributes to p53-mediated apoptosis. Mol Cell. 2007;26:731–43.PubMedCrossRefGoogle Scholar
  24. 24.
    Sun F, Fu H, Liu Q, Tie Y, Zhu J, Xing R, et al. Downregulation of CCND1 and CDK6 by miR-34a induces cell cycle arrest. FEBS Lett. 2008;582:1564–8.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Mojin Wang
    • 1
  • Peng Zhang
    • 1
  • Yuan Li
    • 2
  • Guanghui Liu
    • 1
  • Bin Zhou
    • 1
  • Lan Zhan
    • 1
  • Zongguang Zhou
    • 1
  • Xiaofeng Sun
    • 3
  1. 1.Department of Gastrointestinal Surgery, Institute of Digestive Surgery and National Key Laboratory of Biotherapy of West China HospitalSichuan UniversityChengduChina
  2. 2.Department of Pediatric Surgery, West China HospitalSichuan UniversityChengduChina
  3. 3.Department of Oncology, Institute of Clinical and Experimental MedicineUniversity of LinköpingLinköpingSweden

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