Digestive Diseases and Sciences

, Volume 59, Issue 2, pp 336–345 | Cite as

miR-449a Regulates Proliferation and Chemosensitivity to Cisplatin by Targeting Cyclin D1 and BCL2 in SGC7901 Cells

  • Jianghong Hu
  • Yue Fang
  • Yuan Cao
  • Rong Qin
  • Qiaoyun Chen
Original Article



Recently, several miRNAs have been determined as tumor suppressors in various cancers, such as microRNA-449a. However, the exact molecular mechanisms underlying miR-449a regulated cell proliferation and chemosensitivity in gastric cancer cells have not been well documented.


The present study was designed to test whether miR-449a mediates cell proliferation and chemosensitivity in gastric cancer cells via regulating cyclin D1 and BCL2.


In vitro, the ability of cell proliferation and cell viability were measured by MTT assay; cell cycle and cell apoptosis was detected by FCM. qRT-PCR was used to measure the expression of miR-449a. Western blot and real-time PCR assays were used to detect the expression of cyclin D1 and BCL2 in gastric cancer cell line SGC7901.


miR-449a expression was downregulated in gastric cancer cell line SGC7901 and human gastric cancer tissues, compared to the gastric epithelial cell line GES-1 and matched non-tumor associated tissues. Upregulation of miR-449a reduced the proliferation of SGC7901 cells. Ectopic expression of miR-449a decreased the percentage of S phase cells, increased the percentage of G1/G0 phase cells and increased the apoptosis induced by cisplatin. Moreover, miR-449a inhibited SGC7901 cells proliferation and enhanced cisplatin chemosensitivity by downregulating expression of BCL2 and cyclin D1, respectively, via directly targeting the 3′-untranslated regions of BCL2 and cyclin D1 mRNA.


This is the first report to provide evidence that miR-449a could modulate cell cycle and apoptosis through regulating cyclin D1 and BCL2 expression in SGC7901 cells.


miR-449a BCL2 protein Cyclin D1 protein Gastric cancer 


Conflict of interest



  1. 1.
    Jemal A, Siegel R, Ward E, et al. Cancer statistics, 2006. CA Cancer J Clin. 2006;56:106–130.PubMedCrossRefGoogle Scholar
  2. 2.
    Dicken BJ, Bigam DL, Cass C, et al. Gastric adenocarcinoma: review and considerations for future directions. Ann Surg. 2005;241:27–39.PubMedGoogle Scholar
  3. 3.
    Cunningham D, Allum WH, Stenning SP, et al. Perioperative chemotherapy versus surgery alone for resectable gastroesophageal cancer. N Engl J Med. 2006;355:11–20.PubMedCrossRefGoogle Scholar
  4. 4.
    Liu K, Qian T, Tang L, et al. Decreased expression of microRNA let-7i and its association with chemotherapeutic response in human gastric cancer. World J Surg Oncol. 2012;10:225.PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Ambros V. The functions of animal microRNAs. Nature. 2004;431:350–355.PubMedCrossRefGoogle Scholar
  6. 6.
    de Moor CH, Meijer H, Lissenden S. Mechanisms of translational control by the 3′ UTR in development and differentiation. Semin Cell Dev Biol. 2005;16:49–58.PubMedCrossRefGoogle Scholar
  7. 7.
    Kovalchuk O, Filkowski J, Meservy J, et al. Involvement of microRNA-451 in resistance of the MCF-7 breast cancer cells to chemotherapeutic drug doxorubicin. Mol Cancer Ther. 2008;7:2152–2159.PubMedCrossRefGoogle Scholar
  8. 8.
    Boren T, Xiong Y, Hakam A, et al. MicroRNAs and their target messenger RNAs associated with ovarian cancer response to chemotherapy. Gynecol Oncol. 2009;113:249–255.PubMedCrossRefGoogle Scholar
  9. 9.
    Ji Q, Hao X, Meng Y, et al. Restoration of tumor suppressor miR-34 inhibits human p53-mutant gastric cancer tumorspheres. BMC Cancer. 2008;8:266.PubMedCentralPubMedCrossRefGoogle Scholar
  10. 10.
    Yang N, Kaur S, Volinia S, et al. MicroRNA microarray identifies let-7i as a novel biomarker and therapeutic target in human epithelial ovarian cancer. Cancer Res. 2008;68:10307–10314.PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Noonan EJ, Place RF, Pookot D, et al. miR-449a targets HDAC-1 and induces growth arrest in prostate cancer. Oncogene. 2009;28:1714–1724.PubMedCrossRefGoogle Scholar
  12. 12.
    Lize M, Pilarski S, Dobbelstein M. E2F1-inducible microRNA 449a/b suppresses cell proliferation and promotes apoptosis. Cell Death Differ. 2010;17:452–458.PubMedCrossRefGoogle Scholar
  13. 13.
    Bou Kheir T, Futoma-Kazmierczak E, Jacobsen A, et al. miR-449 inhibits cell proliferation and is down-regulated in gastric cancer. Mol Cancer. 2011;10:29.PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Capuano M, Iaffaldano L, Tinto N, et al. MicroRNA-449a overexpression, reduced NOTCH1 signals and scarce goblet cells characterize the small intestine of celiac patients. PLoS One. 2011;6:e29094.PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Biliran H Jr, Wang Y, Banerjee S, et al. Overexpression of cyclin D1 promotes tumor cell growth and confers resistance to cisplatin-mediated apoptosis in an elastase-myc transgene-expressing pancreatic tumor cell line. Clin Cancer Res Off J Am Assoc Cancer Res. 2005;11:6075–6086.CrossRefGoogle Scholar
  16. 16.
    Dai ZJ, Gao J, Ji ZZ, et al. Matrine induces apoptosis in gastric carcinoma cells via alteration of Fas/FasL and activation of caspase-3. J Ethnopharmacol. 2009;123:91–96.PubMedCrossRefGoogle Scholar
  17. 17.
    Lu J, Getz G, Miska EA, et al. MicroRNA expression profiles classify human cancers. Nature. 2005;435:834–838.PubMedCrossRefGoogle Scholar
  18. 18.
    Iorio MV, Croce CM. MicroRNAs in cancer: small molecules with a huge impact. J Clinical Oncol Off J Am Soc ClinOncol. 2009;27:5848–5856.CrossRefGoogle Scholar
  19. 19.
    Chen H, Lin YW, Mao YQ, et al. MicroRNA-449a acts as a tumor suppressor in human bladder cancer through the regulation of pocket proteins. Cancer Lett. 2012;320:40–47.PubMedCrossRefGoogle Scholar
  20. 20.
    Yang X, Feng M, Jiang X, et al. miR-449a and miR-449b are direct transcriptional targets of E2F1 and negatively regulate pRb-E2F1 activity through a feedback loop by targeting CDK6 and CDC25A. Genes Dev. 2009;23:2388–2393.PubMedCrossRefGoogle Scholar
  21. 21.
    Fu M, Wang C, Li Z, et al. Minireview: cyclin D1: normal and abnormal functions. Endocrinology. 2004;145:5439–5447.PubMedCrossRefGoogle Scholar
  22. 22.
    Schultz J, Lorenz P, Gross G, et al. MicroRNA let-7b targets important cell cycle molecules in malignant melanoma cells and interferes with anchorage-independent growth. Cell Res. 2008;18:549–557.PubMedCrossRefGoogle Scholar
  23. 23.
    Yang J, Liu X, Bhalla K, et al. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science. 1997;275:1129–1132.PubMedCrossRefGoogle Scholar
  24. 24.
    Xia L, Zhang D, Du R, et al. miR-15b and miR-16 modulate multidrug resistance by targeting BCL2 in human gastric cancer cells. Int J Cancer. 2008;123:372–379.PubMedCrossRefGoogle Scholar
  25. 25.
    Zhu W, Shan X, Wang T, et al. miR-181b modulates multidrug resistance by targeting BCL2 in human cancer cell lines. Int J Cancer. 2010;127:2520–2529.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Jianghong Hu
    • 1
  • Yue Fang
    • 2
  • Yuan Cao
    • 2
  • Rong Qin
    • 3
  • Qiaoyun Chen
    • 4
  1. 1.Department of DigestionThe Danyang People’s HospitalDanyangPeople’s Republic of China
  2. 2.Department of Central Laboratory, The Fourth Affiliated People’s HospitalJiangsu UniversityZhenjiangPeople’s Republic of China
  3. 3.Department of Oncology, The Affiliated of People’s HospitalJiangsu UniversityZhenjiangPeople’s Republic of China
  4. 4.Department of Central Laboratory, The Affiliated People’s HospitalJiangsu UniversityZhenjiangPeople’s Republic of China

Personalised recommendations