Skip to main content

Advertisement

SpringerLink
Log in

Expression and regulatory function of miRNA-34a in targeting survivin in gastric cancer cells

  • Research Article
  • Published:
Tumor Biology

Abstract

We aimed to investigate the expression of microRNA-34a (miR-34a) in human gastric cancer cells and to evaluate the effects of miR-34a, acting via its gene survivin, on gastric cancer cell HGC-27 to provide potential new strategies for treating gastric cancer. In vitro cultures of the human gastric cancer cell lines MGC80-3, HGC-27, NCI-N87, and SGC-7901 and the normal human gastric epithelial cell line GES-1 were established. The expression of miR-34a in each gastric cancer cell line and GES-1 normal human gastric epithelial cell line was detected using quantitative real-time polymerase chain reaction (qRT-PCR). After the HGC-27 cells were transfected with a miR-34a mimic for 48 h, the changes in the expression levels of miR-34a were detected using qRT-PCR. The effect of miR-34a on HGC-27 cell viability was measured using a tetrazolium-based colorimetric [−(4,5)-dimethylthiahiazo-(−z-y1)-3,5-di-phenytetrazoliumromide (MTT)] assay. Flow cytometry was used to analyze the effects of miR-34a on HGC-27 cell proliferation. Annexin V/propidium iodide double staining and flow cytometry were used to analyze the effects of miR-34a on HGC-27 cell apoptosis. A Transwell invasion chamber was used to detect the effects of miR-34a on HGC-27 cell invasion. Finally, western blotting was used to analyze the effects of miR-34a on survivin protein expression. The qRT-PCR test determined that miR-34a expression in gastric cancer cells was significantly reduced compared to the normal gastric epithelial cell line GES-1 (p < 0.01). Compared to the control group, cellular miR-34a expression levels were significantly increased in HGC-27 human gastric carcinoma cells after transfection with a miR-34a mimic for 48 h (p < 0.01). The MTT assay demonstrated that after overexpressing miR-34a in HGC-27 cells, cellular viability was significantly reduced (p < 0.05). Flow cytometry analysis determined that upon miR-34a overexpression, the proliferation index decreased significantly (p < 0.05), and cellular apoptosis was significantly increased (p < 0.01). The Transwell invasion chamber assay illustrated that after increasing the expression of miR-34a, the number of cells passing through the Transwell chamber was significantly reduced (p < 0.01). Based on western blotting, compared with the control group, survivin protein expression levels were significantly decreased in the HGC-27 cells transfected with the miR-34a mimic for 48 h (p < 0.01). In conclusion, the expression level of miR-34a was downregulated in human gastric cancer cell lines. miR-34a can negatively regulate survivin protein expression and inhibit gastric cancer cell proliferation and invasion. Therapeutically enhancing miR-34a expression or silencing the survivin gene may benefit patients with gastric cancer.

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
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Shan F, Ji J. Updating advances and controversies on the multidisciplinary therapy of gastric cancer. Transl Gastrointest Cancer. 2012;1(2):151–60.

    Google Scholar 

  2. Hu B, El Hajj N, Sittler S, Lammert N, Barnes R, Meloni-Ehrig A. Gastric cancer: classification, histology and application of molecular pathology. J Gastrointest Oncol. 2012;3(3):251–61.

    PubMed  Google Scholar 

  3. Zhang XD, Shu YQ, Liang J, Zhang FC, Ma XZ, Huang JJ, et al. Combination chemotherapy with paclitaxel, cisplatin and fluorouracil for patients with advanced and metastatic gastric or esophagogastric junction adenocarcinoma: a multicenter prospective study. Chin J Cancer Res. 2012;24(4):291–8.

    PubMed  Google Scholar 

  4. Yang D, Hendifar A, Lenz C, Togawa K, Lenz F, Lurje G, et al. Survival of metastatic gastric cancer: significance of age, sex and race/ethnicity. J Gastrointest Oncol. 2011;2(2):77–84.

    PubMed  CAS  Google Scholar 

  5. Leja M, Wex T, Malfertheiner P. Markers for gastric cancer premalignant lesions: where do we go? Dig Dis. 2012;30(3):268–76.

    Article  PubMed  Google Scholar 

  6. Hopkins S, Yang GY. FDG PET imaging in the staging and management of gastric cancer. J Gastrointest Oncol. 2011;2(1):39–44.

    PubMed  Google Scholar 

  7. Amedei A, Della Bella C, Silvestri E, Prisco D, D’Elios MM. T cells in gastric cancer: friends or foes. Clin Dev Immunol. 2012;2012:690571.

    Article  PubMed  Google Scholar 

  8. Gigek CO, Chen ES, Calcagno DQ, Wisnieski F, Burbano RR, Smith MA. Epigenetic mechanisms in gastric cancer. Epigenomics. 2012;4(3):279–94.

    Article  PubMed  CAS  Google Scholar 

  9. Sun YF, Yang XR, Zhou J, Qiu SJ, Fan J, Xu Y. Circulating tumor cells: advances in detection methods, biological issues, and clinical relevance. J Cancer Res Clin Oncol. 2011;137(8):1151–73.

    Article  PubMed  Google Scholar 

  10. Achyut BR, Yang L. Transforming growth factor-beta in the gastrointestinal and hepatic tumor microenvironment. Gastroenterology. 2011;141(4):1167–78.

    Article  PubMed  CAS  Google Scholar 

  11. St Hill CA. Interactions between endothelial selectins and cancer cells regulate metastasis. Front Biosci. 2011;17:3233–51.

    Article  Google Scholar 

  12. Ali AS, Ahmad A, Ali S, Bao B, Philip PA, Sarkar FH. The role of cancer stem cells and miRNAs in defining the complexities of brain metastasis. J Cell Physiol. 2013;228(1):36–42.

    Article  PubMed  CAS  Google Scholar 

  13. Meng F, Francis H, Alpini G. Non-coding RNAs in human liver malignancies, critical regulators for cancer stemness? Transl Gastrointest Cancer. 2012;1(1):1–4.

    Google Scholar 

  14. Kong YW, Ferland-McCollough D, Jackson TJ, Bushell M. microRNAs in cancer management. Lancet Oncol. 2012;13(6):e249–58.

    Article  PubMed  CAS  Google Scholar 

  15. Li JH, Zhang SW, Liu J, Shao MZ, Chen L. Review of clinical investigation on recurrence of gastric cancer following curative resection. Chin Med J (Engl). 2012;125(8):1479–95.

    Google Scholar 

  16. Bijelic L, Sugarbaker PH. The role of intraperitoneal chemotherapy in the treatment of patients with advanced gastric cancer. Ann Ital Chir. 2012;83(3):224–31.

    PubMed  Google Scholar 

  17. Link A, Kupcinskas J, Wex T, Malfertheiner P. Macro-role of microRNA in gastric cancer. Dig Dis. 2012;30(3):255–67.

    Article  PubMed  Google Scholar 

  18. Wang X, Cao L, Wang Y, Liu N, You Y. Regulation of let-7 and its target oncogenes (review). Oncol Lett. 2012;3(5):955–60.

    PubMed  CAS  Google Scholar 

  19. Li X, Wang Z. The role of noncoding RNA in thyroid cancer. Gland Surg. 2012;1(3):146–50.

    Google Scholar 

  20. Pasquinelli AE. MicroRNAs and their targets: recognition, regulation and an emerging reciprocal relationship. Nat Rev Genet. 2012;13(4):271–82.

    PubMed  CAS  Google Scholar 

  21. Park SH, Zhu Y, Ozden O, Kim HS, Jiang H, Deng CX, et al. SIRT2 is a tumor suppressor that connects aging, acetylome, cell cycle signaling, and carcinogenesis. Transl Cancer Res. 2012;1(1):15–21.

    PubMed  Google Scholar 

  22. Yang JS, Maurin T, Lai EC. Functional parameters of dicer-independent microRNA biogenesis. RNA. 2012;18(5):945–57.

    Article  PubMed  Google Scholar 

  23. Wang J, Li LC. Small RNA and its application in andrology and urology. Transl Androl Urol. 2012;1(1):33–43.

    PubMed  CAS  Google Scholar 

  24. Quintavalle C, Condorelli G. Dulanermin in cancer therapy: still much to do. Transl Lung Cancer Res. 2012;1(2):158–9.

    Google Scholar 

  25. Yamamoto H, Adachi Y, Taniguchi H, Kunimoto H, Nosho K, Suzuki H, et al. Interrelationship between microsatellite instability and microRNA in gastrointestinal cancer. World J Gastroenterol. 2012;18(22):2745–55.

    Article  PubMed  CAS  Google Scholar 

  26. Wong MY, Yu Y, Walsh WR, Yang JL. microRNA-34 family and treatment of cancers with mutant or wild-type p53 (review). Int J Oncol. 2011;38(5):1189–95.

    PubMed  CAS  Google Scholar 

  27. Ghanem M, Levy Y, Mazeh H. Preoperative diagnosis of benign thyroid nodules with intermediate cytology. Gland Surg. 2012;1(2):89–91.

    Google Scholar 

  28. Cha YH, Kim NH, Park C, Lee I, Kim HS, Yook JI. MiRNA-34 intrinsically links p53 tumor suppressor and Wnt signaling. Cell Cycle. 2012;11(7):1273–81.

    Article  PubMed  CAS  Google Scholar 

  29. Balca-Silva J, Neves SS, Goncalves AC, Abrantes AM, Casalta-Lopes J, Botelho MF, et al. Effect of miR-34b overexpression on the radiosensitivity of non-small cell lung cancer cell lines. Anticancer Res. 2012;32(5):1603–9.

    PubMed  CAS  Google Scholar 

  30. Nalls D, Tang SN, Rodova M, Srivastava RK, Shankar S. Targeting epigenetic regulation of miR-34a for treatment of pancreatic cancer by inhibition of pancreatic cancer stem cells. PLoS One. 2011;6(8):e24099.

    Article  PubMed  CAS  Google Scholar 

  31. Lodygin D, Tarasov V, Epanchintsev A, Berking C, Knyazeva T, Korner H, et al. Inactivation of miR-34a by aberrant CpG methylation in multiple types of cancer. Cell Cycle. 2008;7(16):2591–600.

    Article  PubMed  CAS  Google Scholar 

  32. McKenzie JA, Grossman D. Role of the apoptotic and mitotic regulator survivin in melanoma. Anticancer Res. 2012;32(2):397–404.

    PubMed  CAS  Google Scholar 

  33. Church DN, Talbot DC. Survivin in solid tumors: rationale for development of inhibitors. Curr Oncol Rep. 2012;14(2):120–8.

    Article  PubMed  CAS  Google Scholar 

  34. Kumar B, Yadav A, Lang J, Teknos TN, Kumar P. Dysregulation of microRNA-34a expression in head and neck squamous cell carcinoma promotes tumor growth and tumor angiogenesis. PLoS One. 2012;7(5):e37601.

    Article  PubMed  CAS  Google Scholar 

  35. Calin GA, Dumitru CD, Shimizu M, Bichi R, Zupo S, Noch E, et al. Frequent deletions and down-regulation of micro-RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proc Natl Acad Sci U S A. 2002;99(24):15524–9.

    Article  PubMed  CAS  Google Scholar 

  36. Cho WC. MicroRNAs as therapeutic targets and their potential applications in cancer therapy. Expert Opin Ther Targets. 2012;16(8):747–59.

    Article  PubMed  CAS  Google Scholar 

  37. Izzotti A, Cartiglia C, Steele VE, De Flora S. MicroRNAs as targets for dietary and pharmacological inhibitors of mutagenesis and carcinogenesis. Mutat Res. 2012;751(2):287–303.

    Article  PubMed  CAS  Google Scholar 

  38. Vira D, Basak SK, Veena MS, Wang MB, Batra RK, Srivatsan ES. Cancer stem cells, microRNAs, and therapeutic strategies including natural products. Cancer Metastasis Rev. 2012;31(3–4):733–51.

    Article  PubMed  CAS  Google Scholar 

  39. Yao Y, Suo AL, Li ZF, Liu LY, Tian T, Ni L, et al. MicroRNA profiling of human gastric cancer. Mol Med Report. 2009;2(6):963–70.

    CAS  Google Scholar 

  40. Wong KY, Yu L, Chim CS. DNA methylation of tumor suppressor miRNA genes: a lesson from the miR-34 family. Epigenomics. 2011;3(1):83–92.

    Article  PubMed  CAS  Google Scholar 

  41. Shen Z, Zhan G, Ye D, Ren Y, Cheng L, Wu Z, et al. MicroRNA-34a affects the occurrence of laryngeal squamous cell carcinoma by targeting the antiapoptotic gene survivin. Med Oncol. 2012;29(4):2473–80.

    Article  PubMed  CAS  Google Scholar 

  42. Sun L, Wu Z, Shao Y, Pu Y, Miu W, Yao J, et al. MicroRNA-34a suppresses cell proliferation and induces apoptosis in U87 glioma stem cells. Technol Cancer Res Treat. 2012;11(5):483–90.

    PubMed  CAS  Google Scholar 

  43. Li L, Yuan L, Luo J, Gao J, Guo J, Xie X. MiR-34a inhibits proliferation and migration of breast cancer through down-regulation of Bcl-2 and SIRT1. Clin Exp Med. 2012. doi:10.1007/s10238-012-0186-5

Download references

Conflicts of interest

The authors have no commercial, proprietary, or financial interest in the products or companies described in this article.

Author information

Authors and Affiliations

  1. Department of Oncology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

    Weiguo Cao, Hao Li, Jinsong Jiang, Mei Geng & Yening Jin

  2. Department of Gastroenterology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, No. 197, Ruijin Er Rd, Shanghai, 200025, China

    Weiguo Cao, Rong Fan, Lifu Wang, Shidan Cheng & Yunlin Wu

Authors
  1. Weiguo Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rong Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lifu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shidan Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hao Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jinsong Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mei Geng
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yening Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yunlin Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yunlin Wu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cao, W., Fan, R., Wang, L. et al. Expression and regulatory function of miRNA-34a in targeting survivin in gastric cancer cells. Tumor Biol. 34, 963–971 (2013). https://doi.org/10.1007/s13277-012-0632-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13277-012-0632-8

Keywords

Search

Navigation