miR-224 promotion of cell migration and invasion by targeting Homeobox D 10 gene in human hepatocellular carcinoma
Qiong Li
Experimental Center of Basic Medicine, College of Basic Medical Science, Third Military Medical University, Chongqing, China
These authors contributed equally to this work.Search for more papers by this authorChenchen Ding
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
These authors contributed equally to this work.Search for more papers by this authorChuan Chen
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorZhimin Zhang
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorHe Xiao
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorFei Xie
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorLin Lei
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorYuanyuan Chen
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorBijing Mao
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorMei Jiang
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorJian Li
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorDong Wang
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorCorresponding Author
Ge Wang
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Correspondence
Professor Ge Wang, Cancer Center, Daping Hospital & Research Institute of Field Surgery, Third Military Medical University, Chongqing 400042, China. Email: [email protected]
Search for more papers by this authorQiong Li
Experimental Center of Basic Medicine, College of Basic Medical Science, Third Military Medical University, Chongqing, China
These authors contributed equally to this work.Search for more papers by this authorChenchen Ding
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
These authors contributed equally to this work.Search for more papers by this authorChuan Chen
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorZhimin Zhang
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorHe Xiao
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorFei Xie
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorLin Lei
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorYuanyuan Chen
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorBijing Mao
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorMei Jiang
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorJian Li
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorDong Wang
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Search for more papers by this authorCorresponding Author
Ge Wang
Cancer Center, Daping Hospital & Institute of Research Institute of Field Surgery, Third Military Medical University, Chongqing, China
Correspondence
Professor Ge Wang, Cancer Center, Daping Hospital & Research Institute of Field Surgery, Third Military Medical University, Chongqing 400042, China. Email: [email protected]
Search for more papers by this authorAbstract
Background and Aim
MicroRNAs (miRNAs) are small noncoding RNA molecules that control target gene expression and are implicated in the regulation of diverse cellular pathways. In our previous research, we have demonstrated that miR-224 was overexpressed in liver cancer cells and tissues, which was an important factor in the regulation of cell migration and invasion. This study aimed to further explore the regulatory mechanism of miR-224 in the migration and invasion in liver cancer cells.
Methods
A luciferase reporter assay was used to confirm that the HOXD10 gene was a direct target of miR-224. Quantitative reverse transcriptase-polymerase chain reaction, Western blotting, Transwell migration, and Matrigel invasion assays were performed to clarify the molecular mechanism of miR-224 in the regulation of cell migration and invasion in human hepatocellular carcinoma (HCC).
Results
(i) The expression of miR-224 was strongly upregulated in MHHC97H and MHCC97L cells, and its expression level was significantly associated with cell invasive potential. (ii) The HOXD10 gene was confirmed to be a direct target of miR-224. Compared with normal liver tissues and cells, HOXD10 had lower expression in HCC tissues and cells and inversely regulated HCC cell invasion. (iii) miR-224 promoted expression of the tumor invasion-associated proteins p-PAK4 and MMP-9 by directly targeting HOXD10.
Conclusion
Our findings suggest a previously undescribed regulatory pathway in which the miR-224/HOXD10/p-PAK4/MMP-9 signaling pathway contributes to the regulation of cell migration and invasion and provides a new biotarget for HCC treatment.
References
- 1 Lai ECH, Lau WY. The continuing challenge of hepatic cancer in Asia. Surgeon 2005; 3: 210–215.
- 2 Calin GA, Croce CM. MicroRNA signatures in human cancers. Nat. Rev. Cancer 2006; 6: 857–866.
- 3 Zhang B, Pan X, Cobb GP, Anderson TA. MicroRNAs as oncogenes and tumor suppressors. Dev. Biol. 2007; 302: 1–12.
- 4 Gramantieri L, Fornari F, Callegari E et al. MicroRNA involvement in hepatocellular carcinoma. J. Cell. Mol. Med. 2008; 12: 2189–2204.
- 5 Coulouarn C, Factor VM, Andersen JB, Durkin ME, Thorgeirsson SS. Loss of miR-122 expression in liver cancer correlates with suppression of the hepatic phenotype and gain of metastatic properties. Oncogene 2009; 28: 3526–3536.
- 6 Li N, Fu H, Tie Y et al. miR-34a inhibits migration and invasion by down-regulation of c-Met expression in human hepatocellular carcinoma cells. Cancer Lett. 2009; 275: 44–53.
- 7 Meng F, Henson R, Wehbe-Janek H, Ghoshal K, Jacob ST, Patel T. MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology 2007; 133: 647–658.
- 8 Lin S-L, Miller JD, Ying S-Y. Intronic MicroRNA (mi RNA). BioMed Res. Int. 2006; 2006: 26818.
- 9 Zhang Y, He B-S, Pan Y-Q, Xu Y-Q, Wang S-K. Association of OGG1 Ser326Cys polymorphism with colorectal cancer risk: a meta-analysis. Int. J Colorectal Dis. 2011; 26: 1525–1530.
- 10 Zhang Y, Takahashi S, Tasaka A, Yoshima T, Ochi H, Chayama K. Involvement of microRNA-224 in cell proliferation, migration, invasion and anti-apoptosis in hepatocellular carcinoma. J. Gastroenterol. Hepatol. 2013; 28: 565–575.
- 11 Wang Y, Lee ATC, Ma JZI et al. Profiling microRNA expression in hepatocellular carcinoma reveals microRNA-224 up-regulation and apoptosis inhibitor-5 as a microRNA-224-specific target. J. Biol. Chem. 2008; 283: 13205–13215.
- 12 Yao G, Yin M, Lian J et al. MicroRNA-224 is involved in transforming growth factor-β-mediated mouse granulosa cell proliferation and granulosa cell function by targeting Smad4. Mol. Endocrinol. 2010; 24: 540–551.
- 13 Li Q, Wang G, Shan JL et al. MicroRNA-224 is upregulated in HepG2 cells and involved in cellular migration and invasion. J. Gastroenterol. Hepatol. 2009; 25: 164–171.
- 14 Paliouras GN, Naujokas MA, Park M. Pak4, a novel Gab1 binding partner, modulates cell migration and invasion by the Met receptor. Mol. Cell. Biol. 2009; 29: 3018–3032.
- 15 Mak GW-Y, Chan MM-L, Leong VY-L et al. Overexpression of a novel activator of PAK4, the CDK5 kinase-associated protein CDK5RAP3, promotes hepatocellular carcinoma metastasis. Cancer Res. 2011; 71: 2949–2958.
- 16 Turpeenniemi-Hujanen T. Gelatinases (MMP-2 and-9) and their natural inhibitors as prognostic indicators in solid cancers. Biochimie 2005; 87: 287–297.
- 17 Makiyama K, Hamada J-I, Takada M et al. Aberrant expression of HOX genes in human invasive breast carcinoma. Oncol. Rep. 2005; 13: 673–679.
- 18 Cho M, Yashar P, Kim SK et al. HoxD10 gene delivery using adenovirus/adeno-associate hybrid virus inhibits the proliferation and tumorigenicity of GH4 pituitary lactotrope tumor cells. Biochem. Biophys. Res. Commun. 2008; 371: 371–374.
- 19 Sun L, Yan W, Wang Y et al. MicroRNA-10b induces glioma cell invasion by modulating MMP-14 and uPAR expression via HOXD10. Brain Res. 2011; 1389: 9–18.
- 20 Wang L, Chen S, Xue M et al. Homeobox D10 gene, a candidate tumor suppressor, is downregulated through promoter hypermethylation and associated with gastric carcinogenesis. Mol. Med. 2012; 18: 389–400.
- 21 Tian J, Tang ZY, Ye SL et al. New human hepatocellular carcinoma (HCC) cell line with highly metastatic potential (MHCC97) and its expressions of the factors associated with metastasis. Br. J. Cancer 1999; 81: 814–821.
- 22 Li Y, Tang Z-Y, Ye S-L et al. Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97. World J. Gastroenterol. 2001; 7: 630–636.
- 23 Wu Q-f, Liu C, Tai M-h et al. Knockdown of FoxM1 by siRNA interference decreases cell proliferation, induces cell cycle arrest and inhibits cell invasion in MHCC-97H cells in vitro. Acta Pharmacol. Sin. 2010; 31: 361–366.
- 24 Mencia N, Selga E, Noé V. Underexpression of miR-224 in methotrexate resistant human colon cancer cells. Biochem. Pharmacol. 2011; 82: 1572–1582.
- 25 Mees ST, Mardin WA, Sielker S et al. Involvement of CD40 targeting miR-224 and miR-486 on the progression of pancreatic ductal adenocarcinomas. Ann. Surg. Oncol. 2009; 16: 2339–2350.
- 26 Prueitt RL, Yi M, Hudson RS et al. Expression of microRNAs and protein-coding genes associated with perineural invasion in prostate cancer. Prostate 2008; 68: 1152–1164.
- 27 Nikiforova MN, Tseng GC, Steward D, Diorio D, Nikiforov YE. MicroRNA expression profiling of thyroid tumors: biological significance and diagnostic utility. J. Clin. Endocrinol. Metab. 2008; 93: 1600–1608.
- 28 Barber BA, Rastegar M. Epigenetic control of Hox genes during neurogenesis, development, and disease. Ann. Anat. 2010; 192: 261–274.
- 29 Bitu CC, Destro MFSS, Carrera M et al. HOXA1 is overexpressed in oral squamous cell carcinomas and its expression is correlated with poor prognosis. BMC Cancer 2012; 12: 146–156.
- 30 Di Vinci A, Brigati C, Casciano I et al. HOXA7, 9,and 10 are methylation targets associated with aggressive behavior in meningiomas. Transl. Res. 2012; 160: 355–362.
- 31 Shah CA, Bei L, Wang H, Platanias LC, Eklund EA. HoxA10 protein regulates transcription of gene encoding fibroblast growth factor 2 (FGF2) in myeloid cells. J. Biol. Chem. 2012; 287: 18230–18248.
- 32 Yekta S, Tabin CJ, Bartel DP. MicroRNAs in the Hox network: an apparent link to posterior prevalence. Nat. Rev. Genet. 2008; 9: 789–796.
- 33 Yekta S, Shih I, Bartel DP. MicroRNA-directed cleavage of HOXB8 mRNA. Sci. Signal. 2004; 304: 594–596.
- 34 Li Q, Zhu F, Chen P. miR-7 and miR-218 epigenetically control tumor suppressor genes RASSF1A and Claudin-6 by targeting HoxB3 in breast cancer. Biochem. Biophys. Res. Commun. 2012; 424: 28–33.
- 35 Carrio M, Arderiu G, Myers C, Boudreau NJ. Homeobox D10 induces phenotypic reversion of breast tumor cells in a three-dimensional culture model. Cancer Res. 2005; 65: 7177–7185.
- 36 Liu Z, Zhu J, Cao H, Ren H, Fang X. miR-10b promotes cell invasion through RhoC-AKT signaling pathway by targeting HOXD10 in gastric cancer. Int. J. Oncol. 2012; 40: 1560–1567.
- 37 Steeg PS. Cancer: micromanagement of metastasis. Nature 2007; 449: 671–673.
- 38 Clark EA, Golub TR, Lander ES, Hynes RO. Genomic analysis of metastasis reveals an essential role for RhoC. Nature 2000; 406: 532–535.
- 39 Zha Y, Ding E, Yang L et al. Functional dissection of HOXD cluster genes in regulation of neuroblastoma cell proliferation and differentiation. PloS ONE 2012; 7: e40728.
- 40 Ye DZ, Field J. PAK signaling in cancer. Cell. Logistics 2012; 2: 105–116.
- 41 Zhang H-J, Siu MKY, Yeung MCW et al. Overexpressed PAK4 promotes proliferation, migration and invasion of choriocarcinoma. Carcinogenesis 2011; 32: 765–771.
- 42 Kaneyoshi T, Nakatsukasa H, Higashi T et al. Actual invasive potential of human hepatocellular carcinoma revealed by in situ gelatin zymography. Clin. Cancer Res. 2001; 7: 4027–4032.
Citing Literature
