Upregulation of RECK gene expression by small double-stranded RNA targeting the promoter region

Article metrics

Abstract

Recent studies have demonstrated that small double-stranded RNAs (dsRNAs) complementary to the promoter region of target genes enhance the expression of those genes following transfection into cells. Here we show that expression of the matrix metalloproteinase (MMP) inhibitor RECK is activated in the cultured tumor cell lines by transfection with dsRNA complementary to the promoter of the RECK gene, leading to suppression of the expression of MMPs and it inhibited tumor cell invasion. These results support the suggestion that dsRNA complementary to the promoter region of tumor suppressor genes would have potential as a novel antitumor agent.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

References

  1. 1

    Janowski BA, Huffman KE, Schwartz JC, Ram R, Hardy D, Shames DS et al. Inhibiting gene expression at transcription start sites in chromosomal DNA with antigene RNAs. Nat Chem Biol 2005; 1: 216–222.

  2. 2

    Janowski BA, Huffman KE, Schwartz JC, Ram R, Nordsell R, Shames DS et al. Involvement of AGO1 and AGO2 in mammalian transcriptional silencing. Nat Struct Mol Biol 2006; 13: 787–792.

  3. 3

    Morris KV, Chan SW, Jacobsen SE, Looney DJ . Small interfering RNA-induced transcriptional gene silencing in human cells. Science 2004; 305: 1289–1292.

  4. 4

    Ting AH, Schuebel KE, Herman JG, Baylin SB . Short double-stranded RNA induces transcriptional gene silencing in human cancer cells in the absence of DNA methylation. Nat Genet 2005; 37: 906–910.

  5. 5

    Mehndiratta M, Palanichamy JK, Pal A, Bhagat M, Singh A, Sinha S et al. CpG hypermethylation of the C-myc promoter by dsRNA results in growth suppression. Mol Pharm 2011; 8: 2302–2309.

  6. 6

    Hawkins PG, Santoso S, Adams C, Anest V, Morris KV . Promoter targeted small RNAs induce long-term transcriptional gene silencing in human cells. Nucleic Acids Res 2009; 37: 2984–2995.

  7. 7

    Janowski BA, Younger ST, Hardy DB, Ram R, Huffman KE, Corey DR . Activating gene expression in mammalian cells with promoter-targeted duplex RNAs. Nat Chem Biol 2007; 3: 166–173.

  8. 8

    Matsui M, Sakurai F, Elbashir S, Foster DJ, Manoharan M, Corey DR . Activation of LDL receptor expression by small RNAs complementary to a noncoding transcript that overlaps the LDLR promoter. Chem Biol 2010; 17: 1344–1355.

  9. 9

    Huang V, Place RF, Portnoy V, Wang J, Qi Z, Jia Z et al. Upregulation of Cyclin B1 by miRNA and its implications in cancer. Nucleic Acids Res 2012; 40: 1695–1707.

  10. 10

    Kosaka M, Kang MR, Yang G, Li LC . Targeted p21WAF1/CIP1 activation by RNAa inhibits hepatocellular carcinoma cells. Nucleic Acid Ther 2012; 22: 335–343.

  11. 11

    Hu J, Chen Z, Xia D, Wu J, Xu H, Ye ZQ . Promoter-associated small double-stranded RNA interacts with heterogeneous nuclear ribonucleoprotein A2/B1 to induce transcriptional activation. Biochem J 2012; 447: 407–416.

  12. 12

    Yang K, Shen J, Xie YQ, Lin YW, Qin J, Mao QQ et al. Promoter-targeted double-stranded small RNAs activate PAWR gene expression in human cancer cells. Int J Biochem Cell Biol 2013; 45: 1338–1346.

  13. 13

    Li LC, Okino ST, Zhao H, Pookot D, Place RF, Urakami S et al. Small dsRNAs induce transcriptional activation in human cells. Proc Natl Acad Sci USA 2006; 103: 17337–17342.

  14. 14

    Oh J, Takahashi R, Kondo S, Mizoguchi A, Adachi E, Sasahara RM et al. The membrane-anchored MMP inhibitor RECK is a key regulator of extracellular matrix integrity and angiogenesis. Cell 2001; 107: 789–800.

  15. 15

    Takahashi C, Sheng Z, Horan TP, Kitayama H, Maki M, Hitomi K et al. Regulation of matrix metalloproteinase-9 and inhibition of tumor invasion by the membrane-anchored glycoprotein RECK. Proc Natl Acad Sci USA 1998; 95: 13221–13226.

  16. 16

    Liu LT, Chang HC, Chiang LC, Hung WC . Histone deacetylase inhibitor up-regulates RECK to inhibit MMP-2 activation and cancer cell invasion. Cancer Res 2003; 63: 3069–3072.

  17. 17

    Reynolds A, Leake D, Boese Q, Scaringe S, Marshall WS, Khvorova A . Rational siRNA design for RNA interference. Nat Biotechnol 2004; 22: 326–330.

  18. 18

    Chu Y, Yue X, Younger ST, Janowski BA, Corey DR . Involvement of argonaute proteins in gene silencing and activation by RNAs complementary to a non-coding transcript at the progesterone receptor promoter. Nucleic Acids Res 2010; 38: 7736–7748.

  19. 19

    Takagi S, Simizu S, Osada H . RECK negatively regulates matrix metalloproteinase-9 transcription. Cancer Res 2009; 69: 1502–1508.

  20. 20

    Huang V, Qin Y, Wang J, Wang X, Place RF, Lin G et al. RNAa is conserved in mammalian cells. PLoS One 2010; 5: e8848.

  21. 21

    Watts JK, Yu D, Charisse K, Montaillier C, Potier P, Manoharan M et al. Effect of chemical modifications on modulation of gene expression by duplex antigene RNAs that are complementary to non-coding transcripts at gene promoters. Nucleic Acids Res 2010; 38: 5242–5259.

  22. 22

    Place RF, Wang J, Noonan EJ, Meyers R, Manoharan M, Charisse K et al. Formulation of small activating RNA into lipidoid nanoparticles inhibits xenograft prostate tumor growth by inducing p21 expression. Mol Ther Nucleic Acids 2012; 1: e15.

  23. 23

    Kang MR, Yang G, Place RF, Charisse K, Epstein-Barash H, Manoharan M et al. Intravesical delivery of small activating RNA formulated into lipid nanoparticles inhibits orthotopic bladder tumor growth. Cancer Res 2012; 72: 5069–5079.

  24. 24

    Chang HC, Cho CY, Hung WC . Downregulation of RECK by promoter methylation correlates with lymph node metastasis in non-small cell lung cancer. Cancer Sci 2007; 98: 169–173.

  25. 25

    Pesta M, Kulda V, Topolcan O, Safranek J, Vrzalova J, Cerny R et al. Significance of methylation status and the expression of RECK mRNA in lung tissue of patients with NSCLC. Anticancer Res 2009; 29: 4535–4539.

  26. 26

    Schwartz JC, Younger ST, Nguyen NB, Hardy DB, Monia BP, Corey DR et al. Antisense transcripts are targets for activating small RNAs. Nat Struct Mol Biol 2008; 15: 842–848.

  27. 27

    Morris KV, Santoso S, Turner AM, Pastori C, Hawkins PG . Bidirectional transcription directs both transcriptional gene activation and suppression in human cells. PLoS Genet 2008; 4: e1000258.

Download references

Acknowledgements

This study was supported by the Grant-in-Aid for challenging Exploratory Research from the Japan Society of Promotion of Sciences, grants from the Ministry of Health, Labor and Welfare of Japan, and a grant from Suzuken Memorial foundation (to FS).

Author information

Correspondence to F Sakurai.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Sakurai, F., Nanjo, Y., Okamoto, S. et al. Upregulation of RECK gene expression by small double-stranded RNA targeting the promoter region. Cancer Gene Ther 21, 164–170 (2014) doi:10.1038/cgt.2014.12

Download citation

Further reading