Role of SIAH2 protein in the development of hepatocellular carcinoma

doi:10.3877/cma.j.issn.2095-3232.2014.03.014

Abstract

Abstract:

Objective

To investigate the role of SIAH2 protein in the occurrence and development of hepatocellular carcinoma (HCC).

Methods

Human HCC Bel-7404 cells in logarithmic growth phase were inoculated. Empty carrier small interference ribonucleic acid (siRNA) and SIAH2 siRNA were transfected in human HCC Bel-7404 cells. Then the cells were divided into 2 groups: control-siRNA group and SIAH2-siRNA group. The β-actin was taken as control, and the expression of SIAH2 protein in human HCC Bel-7404 cells of 2 groups was detected by Western Blot. The proliferation of cells in 2 groups was detected by methyl thiazolyl tetrazolium (MTT) assay. Cell migration in 2 groups was observed by cell scratch test. Cell invasion in 2 groups was observed by Transwell assay. The data in 2 groups were compared using t test.

Results

The average relative expression of SIAN2 in control-siRNA and SIAH2-siRNA group were 0.71±0.02, 0.33±0.01 respectively. The expression of SIAN2 in SIAH2-siRNA group decreased obviously compared with that in control-siRNA group (t=-4.629, P<0.05). The proliferation rate in SIAH2-siRNA group also decreased obviously. The cell migration rate in SIAH2-siRNA group[(14.3±0.4)% ] was significantly lower than that in control-siRNA group [(45.3±0.4)%]( t=-3.689, P<0.05). The membrane permeating cell count in SIAH2-siRNA group (122±7) was significantly less than that in control-siRNA group (563±10) (t=-3.428, P<0.05).

Conclusion

SIAH2 protein can promote the proliferation, migration and invasion of HCC cells and thus accelerate the occurrence and development of HCC.

Key words: Carcinoma, hepatocellular, Cell proliferation, Cell migration assays, Neoplasm invasiveness, Transfection, Seven in absentia homolog

Ruilei Liu, Jiani Wang, Peng Zhang, Qiaochu Zhang, Xi Li, Hua Jiang, Yong Huang. Role of SIAH2 protein in the development of hepatocellular carcinoma[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2014, 03(03): 189-193.

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Figures/Tables 4

References 16

[1]	符代炎，戴爱国，胡瑞成．细胞氧感受器：天冬酰胺酰羟化酶[J]．中国生物化学与分子生物学报，2007, 23(11):905-910.
[2]	Schmidt RL, Park CH, Ahmed AU, et al. Inhibition of RAS-mediated transformation and tumorigenesis by targeting the downstream E3 ubiquitin ligase seven in absentia homologue[J]. Cancer Res, 2007, 67(24):11798-11810.
[3]	Ahmed AU, Schmidt RL, Park CH, et al. Effect of disrupting seven-in-absentia homolog 2 function on lung cancer cell growth[J]. J Natl Cancer Inst, 2008, 100(22):1606-1629.
[4]	Nakayama K, Frew IJ, Hagensen M, et al. Siah2 regulates stability of prolyl-hydroxylases, controls HIF1alpha abundance, and modulates physiological responses to hypoxia[J]. Cell, 2004, 117(7):941-952.
[5]	Boehm J, He Y, Greiner A, et al. Regulation of BOB.1/OBF.1 stability by SIAH[J]. EMBO J, 2001, 20(15):4153-4162.
[6]	Zhang J, Guenther MG, Carthew RW, et al. Proteasomal regulation of nuclear receptor corepressor-mediated repression[J]. Genes Dev, 1998, 12(12):1775-1780.
[7]	Habelhah H, Frew IJ, Laine A, et al. Stress-induced decrease in TRAF2 stability is mediated by Siah2[J]. EMBO J, 2002, 21(21):5756-5765.
[8]	Jansen MP, Ruigrok-Ritstier K, Dorssers LC, et al. Downregulation of SIAH2, an ubiquitin E3 ligase, is associated with resistance to endocrine therapy in breast cancer[J]. Breast Cancer Res Treat, 2009, 116(2):263-271.
[9]	Shah M, Stebbins JL, Dewing A, et al. Inhibition of Siah2 ubiquitin ligase by vitamin K3 (menadione) attenuates hypoxia and MAPK signaling and blocks melanoma tumorigenesis[J]. Pigment Cell Melanoma Res, 2009, 22(6):799-808.
[10]	Möller A, House CM, Wong CS, et al. Inhibition of Siah ubiquitin ligase function[J]. Oncogene, 2009, 28(2):289-296.
[11]	Nadeau RJ, Toher JL, Yang X, et al. Regulation of Sprouty2 stability by mammalian Seven-in-Absentia homolog 2[J]. J Cell Biochem, 2007, 100(1):151-160.
[12]	Lim J, Yusoff P, Wong ES, et al. The cysteine-rich sprouty translocation domain targets mitogen-activated protein kinase inhibitory proteins to phosphatidylinositol 4,5-bisphosphate in plasma membranes[J]. Mol Cell Biol, 2002, 22(22):7953-7966.
[13]	Gross I, Bassit B, Benezra M, et al. Mammalian sprouty proteins inhibit cell growth and differentiation by preventing ras activation[J]. J Biol Chem, 2001, 276(49):46460-46468.
[14]	Shaw AT, Meissner A, Dowdle JA, et al. Sprouty-2 regulates oncogenic K-ras in lung development and tumorigenesis[J]. Genes Dev, 2007, 21(6):694-707.
[15]	Qi J, Nakayama K, Gaitonde S, et al. The ubiquitin ligase Siah2 regulates tumorigenesis and metastasis by HIF-dependent and -independent pathways[J]. Proc Natl Acad Sci U S A, 2008, 105(43):16713-16718.
[16]	Epstein AC, Gleadle JM, McNeill LA, et al. C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation[J]. Cell, 2001, 107(1):43-54.

组别	次数	划痕宽度（mm）		迁移率（%）
组别	次数	0 h	48 h	迁移率（%）
control-siRNA组	3	26.1±0.9	13.4±0.8	45.3±0.4
SIAH2-siRNA组	3	26.3±0.9	24.8±0.8	14.3±0.4

组别	次数	划痕宽度（mm）		迁移率（%）
组别	次数	0 h	48 h	迁移率（%）
control-siRNA组	3	26.1±0.9	13.4±0.8	45.3±0.4
SIAH2-siRNA组	3	26.3±0.9	24.8±0.8	14.3±0.4

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