Silencing of S100A4 gene inhibits oxygen-induced retinal neovascularization in mice_Chinese Journal of Ocular Fundus Diseases

Authors：

ChengGumeng , HeTao ,  XingYiqiao

Department of Ophthalmology, Renmin Hospital of Wuhan University, Wuhan 430070, China;

Corresponding author：

XingYiqiao, Email: xing_yiqiao@aliyun.com

Keywords：

Retinal neovascularization/prevention & control; Calgranulin A/antagonists & inhibitors; Gene silencing; Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2016.01.013

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the inhibitory effects and related mechanism of S100A4 gene silencing on oxygen-induced retinal neovascularization. Methods 7-day-old C57BL/6J mice were randomly divided into 5 groups including normal group, normal-S100A4 group, oxygen induced retinopathy (OIR) group, OIR-S100A4 group, OIR-green fluorescent protein (GFP) group. To establish the OIR model, mice from all groups except normal one were exposed to (75±2)% oxygen for 5 days and then to room air. In the OIR-S100A4 group and OIR-GFP group, the OIR mice were given an intravitreal injection of 1μl of 1.0×10⁹ PFU/ml adenovirus of Ad-S100A4-RNAi or Ad-GFP at P12, and then returned to normoxia for the next 5 days. In the OIR group, OIR was induced in C57bl/6J mice from P7 to P17. In the normal-S100A4 group, the normal P12 mice were give an intravitreal injection of 1 μl of Ad-S100A4-RNAi adenovirus, and maintained in room air from P12 to P17. In normal group, newborn mouse litters were maintained in room air from P0 to P17 without any treatment. Mice in all five groups were euthanized at P17, and retinas were collected for biochemical assays and morphological study. Retinal neovascularization (RNV) was evaluated by counting the number of pre-retinal neovascular cells and the whole mount immunofluorescent staining of the mouse retina. Protein and mRNA expression levels of S100A4, cAMP responsive element binding protein (CREB), B cell lymphoma-2 (bcl-2), Caspase-3 were determined with western blot and real-time PCR. Results The number of pre-retinal neovascular cell nuclei in retinas from OIR-S100A4 group were obviously lower than those in the retinas from OIR group and OIR-GFP group (t=13.61, 14.64; P < 0.05). In OIR-S100A4 group, the retinal neovascular tufts area and the vaso-oblitertion area were both significantly smaller than those in OIR group and OIR-GFP group (P < 0.05). Protein level of CREB and bcl-2 were significantly down-regulated in OIR-S100A4 group than those in OIR and OIR-GFP group (P < 0.05).On the contrary, protein levels of Caspase-3 were up-regulated in OIR-S100A4 group than those in OIR and OIR-GFP group (P < 0.05). Conclusion Ad-S100A4-RNAi transfer ameliorates RNV in mouse model of OIR maybe through down-regulating the expression of bcl-2 and CREB, and up-regulating the Caspase-3.

Citation： ChengGumeng, HeTao, XingYiqiao. Silencing of S100A4 gene inhibits oxygen-induced retinal neovascularization in mice. Chinese Journal of Ocular Fundus Diseases, 2016, 32(1): 52-57. doi: 10.3760/cma.j.issn.1005-1015.2016.01.013 Copy

1.	Boye K, Maelandsmo GM. S100A4 and metastasis: a small actor playing many roles[J]. Am J Pathol, 2010, 176(2): 528-535.DOI:10.2353/ajpath.2010.090526.
2.	Saleem M, Kweon MH, Johnson JJ, et al. S100A4 accelerates tumorigenesis and invasion of human prostate cancer through the transcriptional regulation of matrix metalloproteinase 9[J]. Proc Natl Acad Sci USA, 2006, 103(40): 14825-14830.
3.	Smith LE, Wesolowski E, McLellan A, et al. Oxygen-induced retinopathy in the mouse[J]. Invest Ophthalmol Vis Sci, 1994, 35(1): 101-111.
4.	Chen YY, Liu SL, Hu DP, et al. N-methyl-N-nitrosourea-induced retinal degeneration in mice[J]. Exp Eye Res, 2014, 121: 102-113. DOI:10.1016/j.exer.2013.12.019.
5.	Ghoul A, Serova M, Astorgues-Xerri L, et al. Epithelial-to-mesenchymal transition and resistance to ingenol 3-angelate, a novel protein kinase C modulator, in colon cancer cells[J]. Cancer Res, 2009, 69(10):4260-4269. DOI: 10.1158/0008-5472.CAN-08-2837.
6.	Malashkevich VN, Varney KM, Garrett SC, et al. Structure of Ca²⁺-bound S100A4 and its interaction with peptides derived from nonmuscle myosin-ⅡA[J]. Biochemistry, 2008, 47(18): 5111-5126. DOI:10.1021/bi702537s.
7.	Kim EJ, Helfman DM. Characterization of the metastasis-associated protein, S100A4:roles of calcium binding and dimerization in cellular localization and interaction with myosin[J]. J Biol Chem, 2003, 278(32): 30063-30073.
8.	Grigorian M, Andresen S, Tulchinsky E, et al. Tumor suppressor p53 protein is a new target for the metastasis-associated Mts1/S100A4 protein: functional consequences of their interaction[J]. J Biol Chem, 2001, 276(25): 22699-22708.
9.	Peshavariya HM, Liu GS, Chang CW, et al. Prostacyclin signaling boosts NADPH oxidase 4 in the endothelium promoting cytoprotection and angiogenesis[J]. Antioxid Redox Signal, 2014, 20(17): 2710-2725.
10.	Darashchonak N, Koepsell B, Bogdanova N, et al. Adenosine A2B receptors induce proliferation, invasion and activation of cAMP response element binding protein (CREB) in trophoblast cells[J]. BMC Pregnancy Childbirth, 2014, 14: 2. DOI: 10.1186/1471-2393-14-2.
11.	Barresi V, Branca G, Caffo M, et al. p-CREB expression in human meningiomas: correlation with angiogenesis and recurrence risk[J]. J Neurooncol, 2015, 122(1): 87-95. DOI: 10.1007/s11060-014-1706-9.
12.	Mayr B, Montminy M. Transcriptional regulation by the phosphorylation-dependent factor CREB[J]. Nat Rev Mol Cell Biol, 2001, 2(8): 599-609.
13.	Meller R, Minami M, Cameron JA, et al. CREB-mediated Bcl-2 protein expression after ischemic preconditioning[J]. J Cereb Blood Flow Metab, 2005, 25(2): 234-246.
14.	Boland K, Flanagan L, Prehn JH. Paracrine control of tissue regeneration and cell proliferation by Caspase-3[J]. Cell Death Dis, 2013, 4: 725. DOI: 10.1038/cddis.2013.250.
15.	Nassar A, Lawson D, Cotsonis G, et al. Survivin and caspase-3 expression in breast cancer: correlation with prognostic parameters, proliferation, angiogenesis, and outcome[J]. Appl Immunohistochem Mol Morphol, 2008, 16(2): 113-120.
16.	Volm M, Mattern J, Koomagi R. Inverse correlation between apoptotic (Fas ligand, caspase-3) and angiogenic factors (VEGF, microvessel density) in squamous cell lung carcinomas[J]. Anticancer Res, 1999, 19(3A): 1669-1671.

1. Boye K, Maelandsmo GM. S100A4 and metastasis: a small actor playing many roles[J]. Am J Pathol, 2010, 176(2): 528-535.DOI:10.2353/ajpath.2010.090526.
2. Saleem M, Kweon MH, Johnson JJ, et al. S100A4 accelerates tumorigenesis and invasion of human prostate cancer through the transcriptional regulation of matrix metalloproteinase 9[J]. Proc Natl Acad Sci USA, 2006, 103(40): 14825-14830.
3. Smith LE, Wesolowski E, McLellan A, et al. Oxygen-induced retinopathy in the mouse[J]. Invest Ophthalmol Vis Sci, 1994, 35(1): 101-111.
4. Chen YY, Liu SL, Hu DP, et al. N-methyl-N-nitrosourea-induced retinal degeneration in mice[J]. Exp Eye Res, 2014, 121: 102-113. DOI:10.1016/j.exer.2013.12.019.
5. Ghoul A, Serova M, Astorgues-Xerri L, et al. Epithelial-to-mesenchymal transition and resistance to ingenol 3-angelate, a novel protein kinase C modulator, in colon cancer cells[J]. Cancer Res, 2009, 69(10):4260-4269. DOI: 10.1158/0008-5472.CAN-08-2837.
6. Malashkevich VN, Varney KM, Garrett SC, et al. Structure of Ca²⁺-bound S100A4 and its interaction with peptides derived from nonmuscle myosin-ⅡA[J]. Biochemistry, 2008, 47(18): 5111-5126. DOI:10.1021/bi702537s.
7. Kim EJ, Helfman DM. Characterization of the metastasis-associated protein, S100A4:roles of calcium binding and dimerization in cellular localization and interaction with myosin[J]. J Biol Chem, 2003, 278(32): 30063-30073.
8. Grigorian M, Andresen S, Tulchinsky E, et al. Tumor suppressor p53 protein is a new target for the metastasis-associated Mts1/S100A4 protein: functional consequences of their interaction[J]. J Biol Chem, 2001, 276(25): 22699-22708.
9. Peshavariya HM, Liu GS, Chang CW, et al. Prostacyclin signaling boosts NADPH oxidase 4 in the endothelium promoting cytoprotection and angiogenesis[J]. Antioxid Redox Signal, 2014, 20(17): 2710-2725.
10. Darashchonak N, Koepsell B, Bogdanova N, et al. Adenosine A2B receptors induce proliferation, invasion and activation of cAMP response element binding protein (CREB) in trophoblast cells[J]. BMC Pregnancy Childbirth, 2014, 14: 2. DOI: 10.1186/1471-2393-14-2.
11. Barresi V, Branca G, Caffo M, et al. p-CREB expression in human meningiomas: correlation with angiogenesis and recurrence risk[J]. J Neurooncol, 2015, 122(1): 87-95. DOI: 10.1007/s11060-014-1706-9.
12. Mayr B, Montminy M. Transcriptional regulation by the phosphorylation-dependent factor CREB[J]. Nat Rev Mol Cell Biol, 2001, 2(8): 599-609.
13. Meller R, Minami M, Cameron JA, et al. CREB-mediated Bcl-2 protein expression after ischemic preconditioning[J]. J Cereb Blood Flow Metab, 2005, 25(2): 234-246.
14. Boland K, Flanagan L, Prehn JH. Paracrine control of tissue regeneration and cell proliferation by Caspase-3[J]. Cell Death Dis, 2013, 4: 725. DOI: 10.1038/cddis.2013.250.
15. Nassar A, Lawson D, Cotsonis G, et al. Survivin and caspase-3 expression in breast cancer: correlation with prognostic parameters, proliferation, angiogenesis, and outcome[J]. Appl Immunohistochem Mol Morphol, 2008, 16(2): 113-120.
16. Volm M, Mattern J, Koomagi R. Inverse correlation between apoptotic (Fas ligand, caspase-3) and angiogenic factors (VEGF, microvessel density) in squamous cell lung carcinomas[J]. Anticancer Res, 1999, 19(3A): 1669-1671.

Chinese Journal of Ocular Fundus Diseases

Silencing of S100A4 gene inhibits oxygen-induced retinal neovascularization in mice

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content