MicroRNA library-based functional screening identified miR-137 as a suppresser of gastric cancer cell proliferation

  • Xiushan Zheng
  • Jiaqiang Dong
  • Taiqian Gong
  • Zhiyong Zhang
  • Ying Wang
  • Yunming Li
  • Yulong Shang
  • Kai Li
  • Gui Ren
  • Bin Feng
  • Juntang Li
  • Qifei Tian
  • Shanhong Tang
  • Li Sun
  • Mengbin Li
  • Hongwei Zhang
  • Daiming Fan
Original Article - Cancer Research

Abstract

Purposes

Uncontrolled proliferation is a key characteristic of gastric carcinogenesis and the precise mechanisms underlying the altered proliferation behaviors of GC cells have not been clearly elucidated. miRNAs has been suggested to play a crucial role in the pathogenesis and development of various cancers. In the present study, we employed an impedance-based real-time cell electronic sensing (RT-CES) system to detect the effects of ectopically expressed miRNAs on GC cell proliferation.

Methods

miRNA mimics were transfected into gastric cancer cell line SGC7901 and the effect of individual miRNA on the proliferation rate of the cells was measured by the RT-CES system. The screening results were validated with qRT-PCR and miR-137 was selected for further research. The effects of ectopically expressed miR-137 on GC cell growth and cell cycle progress were measured using MTT assay and flow cytometry. The target gene of miR-137 was predicted using different bioinformatics tools and the direct interaction between miR-137 and the 3’-UTR was confirmed with a luciferase reporter assay. The in vivo effect of miR-137 on GC cell proliferation was examined with a tumor-bearing nude mouse model. The correlation between miR-137 expression and patients’ prognosis was explored in a cohort of 38 patients. Prognosis was explored in a cohort of 38 patients.

Results

Ectopic expression of miR-137 was sufficient to inhibit GC cell proliferation both in vitro and in vivo. Bioinformatics prediction and luciferase reporter assay revealed CDK6 as a target gene through which miR-137 exerted an inhibitory function. Moreover, miR-137 expression positively correlated with better prognosis.

Conclusion

Our data indicated an important regulatory role of miR-137 in GC cell proliferation and that it may be explored as a prognostic marker for GC.

Keywords

miR-137 Gastric cancer Cell proliferation CDK6 Real-time cell electronic sensing (RT-CES) system 

Supplementary material

432_2014_1847_MOESM1_ESM.doc (312 kb)
Supplementary material 1 (DOC 312 kb)

References

  1. Balaguer F, Link A, Lozano JJ et al (2010) Epigenetic silencing of miR-137 is an early event in colorectal carcinogenesis. Cancer Res 70(16):6609–6618CrossRefPubMedCentralPubMedGoogle Scholar
  2. Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116(2):281–297CrossRefPubMedGoogle Scholar
  3. Bartel DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136(2):215–233CrossRefPubMedCentralPubMedGoogle Scholar
  4. Brito-Babapulle V, Gruszka-Westwood AM, Platt G et al (2002) Translocation t(2;7)(p12;q21-22) with dysregulation of the CDK6 gene mapping to 7q21-22 in a non-Hodgkin’s lymphoma with leukemia. Haematologica 87(4):357–362PubMedGoogle Scholar
  5. Cai CK, Zhao GY, Tian LY et al (2012) miR-15a and miR-16-1 downregulate CCND1 and induce apoptosis and cell cycle arrest in osteosarcoma. Oncol Rep 28(5):1764–1770PubMedGoogle Scholar
  6. Chen DL, Wang DS, Wu WJ et al (2013) Overexpression of paxillin induced by miR-137 suppression promotes tumor progression and metastasis in colorectal cancer. Carcinogenesis 34(4):803–811CrossRefPubMedCentralPubMedGoogle Scholar
  7. Dai L, Wang W, Zhang S et al (2012) Vector-based miR-15a/16-1 plasmid inhibits colon cancer growth in vivo. Cell Biol Int 36(8):765–770CrossRefPubMedGoogle Scholar
  8. Du Y, Liu Z, Gu L et al (2012) Characterization of human gastric carcinoma-related methylation of 9 miR CpG islands and repression of their expressions in vitro and in vivo. BMC Cancer 12:249CrossRefPubMedCentralPubMedGoogle Scholar
  9. Feng L, Xie Y, Zhang H, Wu Y (2012) miR-107 targets cyclin-dependent kinase 6 expression, induces cell cycle G1 arrest and inhibits invasion in gastric cancer cells. Med Oncol 29(2):856–863CrossRefPubMedGoogle Scholar
  10. Gao SM, Xing CY, Chen CQ, Lin SS, Dong PH, Yu FJ (2011) miR-15a and miR-16-1 inhibit the proliferation of leukemic cells by down-regulating WT1 protein level. J Exp Clin Cancer Res 30:110CrossRefPubMedCentralPubMedGoogle Scholar
  11. Grossel MJ, Hinds PW (2006) From cell cycle to differentiation: an expanding role for cdk6. Cell Cycle 5(3):266–270CrossRefPubMedGoogle Scholar
  12. Han Z, Yang Q, Liu B et al (2012) MicroRNA-622 functions as a tumor suppressor by targeting K-Ras and enhancing the anticarcinogenic effect of resveratrol. Carcinogenesis 33(1):131–139CrossRefPubMedGoogle Scholar
  13. Jemal A, Siegel R, Xu J, Ward E (2010) Cancer statistics, 2010. CA Cancer J Clin 60(5):277–300CrossRefPubMedGoogle Scholar
  14. Lang Q, Ling C (2012) MiR-124 suppresses cell proliferation in hepatocellular carcinoma by targeting PIK3CA. Biochem Biophys Res Commun 426(2):247–252CrossRefPubMedGoogle Scholar
  15. Liu M, Lang N, Qiu M et al (2011) miR-137 targets Cdc42 expression, induces cell cycle G1 arrest and inhibits invasion in colorectal cancer cells. Int J Cancer 128(6):1269–1279CrossRefPubMedGoogle Scholar
  16. Luo C, Tetteh PW, Merz PR et al (2013) miR-137 inhibits the invasion of melanoma cells through downregulation of multiple oncogenic target genes. J Invest Dermatol 133(3):768–775CrossRefPubMedGoogle Scholar
  17. Nishida N, Mimori K, Fabbri M et al (2011) MicroRNA-125a-5p is an independent prognostic factor in gastric cancer and inhibits the proliferation of human gastric cancer cells in combination with trastuzumab. Clin Cancer Res 17(9):2725–2733CrossRefPubMedGoogle Scholar
  18. Petrocca F, Visone R, Onelli MR et al (2008) E2F1-regulated microRNAs impair TGFbeta-dependent cell-cycle arrest and apoptosis in gastric cancer. Cancer Cell 13(3):272–286CrossRefPubMedGoogle Scholar
  19. Sawai CM, Freund J, Oh P et al (2012) Therapeutic targeting of the cyclin D3:CDK4/6 complex in T cell leukemia. Cancer Cell 22(4):452–465CrossRefPubMedCentralPubMedGoogle Scholar
  20. Schipper DL, Wagenmans MJ, Peters WH, Wagener DJ (1998) Significance of cell proliferation measurement in gastric cancer. Eur J Cancer 34(6):781–790CrossRefPubMedGoogle Scholar
  21. Solly K, Wang X, Xu X, Strulovici B, Zheng W (2004) Application of real-time cell electronic sensing (RT-CES) technology to cell-based assays. Assay Drug Dev Technol 2(4):363–372CrossRefPubMedGoogle Scholar
  22. Ushijima T, Sasako M (2004) Focus on gastric cancer. Cancer Cell 5(2):121–125CrossRefPubMedGoogle Scholar
  23. Vrba L, Munoz-Rodriguez JL, Stampfer MR, Futscher BW (2013) miRNA gene promoters are frequent targets of aberrant DNA methylation in human breast cancer. PLoS ONE 8(1):e54398CrossRefPubMedCentralPubMedGoogle Scholar
  24. Wang Y, Baskerville S, Shenoy A, Babiarz JE, Baehner L, Blelloch R (2008) Embryonic stem cell-specific microRNAs regulate the G1-S transition and promote rapid proliferation. Nat Genet 40(12):1478–1483CrossRefPubMedCentralPubMedGoogle Scholar
  25. Wang F, Ma YL, Zhang P et al (2013) SP1 mediates the link between methylation of the tumour suppressor miR-149 and outcome in colorectal cancer. J Pathol 229(1):12–24CrossRefPubMedGoogle Scholar
  26. Wu WK, Lee CW, Cho CH et al (2010) MicroRNA dysregulation in gastric cancer: a new player enters the game. Oncogene 29(43):5761–5771CrossRefPubMedGoogle Scholar
  27. Xia J, Wu Z, Yu C et al (2012) miR-124 inhibits cell proliferation in gastric cancer through down-regulation of SPHK1. J Pathol 227(4):470–480CrossRefPubMedGoogle Scholar
  28. Zhang L, Liu X, Jin H et al (2013) miR-206 inhibits gastric cancer proliferation in part by repressing cyclin D2. Cancer Lett 332(1):94–101CrossRefPubMedGoogle Scholar
  29. Zhao Y, Li Y, Lou G et al (2012) MiR-137 targets estrogen-related receptor alpha and impairs the proliferative and migratory capacity of breast cancer cells. PLoS ONE 7(6):e39102CrossRefPubMedCentralPubMedGoogle Scholar
  30. Zhu X, Li Y, Shen H et al (2013a) miR-137 restoration sensitizes multidrug-resistant MCF-7/ADM cells to anticancer agents by targeting YB-1. Acta Biochim Biophys Sin (Shanghai) 45(2):80–86CrossRefGoogle Scholar
  31. Zhu X, Li Y, Shen H et al (2013b) miR-137 inhibits the proliferation of lung cancer cells by targeting Cdc42 and Cdk6. FEBS Lett 587(1):73–81CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Xiushan Zheng
    • 1
    • 2
  • Jiaqiang Dong
    • 1
  • Taiqian Gong
    • 1
    • 3
  • Zhiyong Zhang
    • 1
  • Ying Wang
    • 1
  • Yunming Li
    • 2
  • Yulong Shang
    • 1
  • Kai Li
    • 1
  • Gui Ren
    • 1
  • Bin Feng
    • 1
  • Juntang Li
    • 4
  • Qifei Tian
    • 1
  • Shanhong Tang
    • 1
  • Li Sun
    • 1
  • Mengbin Li
    • 1
  • Hongwei Zhang
    • 1
  • Daiming Fan
    • 1
  1. 1.State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive DiseasesFourth Military Medical UniversityXi’anChina
  2. 2.Department of Thoracic SurgeryGeneral Hospital of Chengdu Military CommandChengduChina
  3. 3.Department of Thoracic Surgery, Daping HospitalThird Military Medical UniversityChongqingChina
  4. 4.State Key Laboratory of Cancer Biology, Department of ImmunologyFourth Military Medical UniversityXi’anChina

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