miR-449a inhibits proliferation and invasion by regulating ADAM10 in hepatocellular carcinoma - PubMed (original) (raw)

. 2016 Jun 15;8(6):2609-19.

eCollection 2016.

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miR-449a inhibits proliferation and invasion by regulating ADAM10 in hepatocellular carcinoma

Songyang Liu et al. Am J Transl Res. 2016.

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Abstract

MicroRNAs (miRNAs) are a class of small, non-coding RNAs that play a crucial role in tumor procession. It has been demonstrated that miR-449a expression was downregulated and served as tumor suppressor in many types of tumor. However, the biological function and molecular mechanism of miR-449a in hepatocellular carcinoma (HCC) still remains largely unknown. Therefore, the aims of this study were to investigate biological role and molecular mechanism of miR-449a in HCC by a serial of molecular experiments. Here, we demonstrated that miR-449a expression was downregulated in HCC tissues and cell lines compared with the adjacent nontumor tissues and normal hepatic cell line. Ectopic expression of miR-449a suppressed HCC cell proliferation, colony formation, migration and invasion. Moreover, A Disintegrin And Metalloproteinases 10 (ADAM10) was identified as a direct target gene of miR-449a in HCC cell. ADAM10 expression was upregulated in HCC tissues and cell lines, and was negatively correlated with the expression level of miR-449a in HCC tissues. Interesting, overexpression of ADAM10 attenuated the inhibition effect of miR-449a-mediated HCC cell proliferation, colony formation, migration and invasion. These results suggested that miR-449a might function as a tumor suppressor miRNA, at least in part, through regulating ADAM10 expression in HCC.

Keywords: ADAM10; Hepatocellular carcinoma; invasion; miR-449a; proliferation.

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Figures

Figure 1

Figure 1

miR-449a expression was downregulated in HCC tissues and cell lines. A. The expression of miR-449a was measured in 40 pairs HCC tissues and adjacent nontumor tissues by qRT-PCR. **P<0.01 versus nontumor tissues. B. The expression of miR-449a was measured in four HCC cell lines (SMMC-7721, Hep3B, HepG2, Huh-7) and normal hepatic cell line HL-7702 by qRT-PCR. *P<0.05, **P<0.01 versus HL-7702.

Figure 2

Figure 2

miR-449a inhibits cell proliferation and colony formation in HCC cells. A. The expression of miR-449a was measured in HepG2 cell after tranfected with miR-449a mimic or miR-NC using qRT-PCR. B. Cell proliferation was determined in HepG2 cell after tranfected with miR-449a mimic or miR-NC using CCK8 assay. C. Colony formation was determined in HepG2 cell after tranfected with miR-449a mimic or miR-NC. *P<0.05, **P<0.01 versus miR-NC.

Figure 3

Figure 3

miR-449a inhibits cell migration and invasion in HCC cells. A. Cell migration was measured in HepG2 cell after tranfected with miR-449a mimic or miR-NC using wound healing assay. B. Cell invasion was determined in HepG2 cell after tranfected with miR-449a mimic or miR-NC using transwell invasion assay. *P<0.05, **P<0.01 versus miR-NC.

Figure 4

Figure 4

miR-449a directly targets ADAM10 3’-UTR. (A) Schematic illustration of the predicted miR-449a-binding sites in ADAM10 3’-UTR. (B) Luciferase activity was measured in HepG2 cells after co-transfected with miR-449a mimic or miR-NC and Wt or Mut ADAM10 3’UTR report plasmid. Wt: wide-type; Mut: mutant-type. (C and D) ADAM10 expression on mRNA level (C) and protein level (D) was measured in HepG2 cell after tranfected with miR-449a mimic or miR-NC by qRT-PCR and western blot, respectively. β-actin was used as a control. *P<0.05, **P<0.01 versus miR-NC.

Figure 5

Figure 5

miR-449a expression is negatively associated with ADAM10 in HCC tissues. A. The mRNA expression of ADAM10 was measured in 40 pairs HCC tissues and adjacent nontumor tissues by qRT-PCR. **P<0.01 versus nontumor tissues. B. ADAM10 protein was measured in HCC tissues and corresponding nontumor tissues by western blot. β-actin was used as a control. C. The negative correlation between ADAM10 mRNA expression and miR-449a expression was determined by Pearson’s correlation. D. The ADAM10 protein expression was measured in four HCC cell lines (SMMC-7721, Hep3B, HepG2, Huh-7) and normal hepatic cell line HL-7702 by western blot. β-actin was used as a control.

Figure 6

Figure 6

ADAM10 rescued the suppressive effect of miR-449a-mediated HCC on cell proliferation, colon formation, and invasion. (A and B) ADAM10 expression on mRNA level (A) and protein level (B) was measured in HepG2 cells transfected with miR-449a with/without ADAM10 overexpression plasmid. β-actin was used as an internal control. (C-F) Cell proliferation (C), colon formation (D), migration (E) and invasion (F) were determined in HepG2 cells transfected with miR-449a with/without ADAM10 overexpression plasmid. *P<0.05, **P<0.01 versus miR-449a.

References

    1. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65:87–108. - PubMed
    1. Mazzola A, Costantino A, Petta S, Bartolotta TV, Raineri M, Sacco R, Brancatelli G, Camma C, Cabibbo G. Recurrence of hepatocellular carcinoma after liver transplantation: an update. Future Oncol. 2015;11:2923–2936. - PubMed
    1. Forner A, Llovet JM, Bruix J. Hepatocellular carcinoma. Lancet. 2012;379:1245–1255. - PubMed
    1. Valinezhad Orang A, Safaralizadeh R, Kazemzadeh-Bavili M. Mechanisms of miRNA-Mediated Gene Regulation from Common Downregulation to mRNA-Specific Upregulation. Int J Genomics. 2014;2014:970607. - PMC - PubMed
    1. Bushati N, Cohen SM. microRNA functions. Annu Rev Cell Dev Biol. 2007;23:175–205. - PubMed

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