Cysteine-rich 61 siRNA reduces retinal neovascularization of mice_Chinese Journal of Ocular Fundus Diseases

Authors：

DiYu , ZhangYiou ,  ChenXiaolong

Department of Ophthalmology, Shengjing Hospital of China Medical University, Shenyang 110004, China;

Corresponding author：

ChenXiaolong, Email: 390532915@qq.com

Keywords：

Retinal neovascularization/drug therapy; Cysteine-rich protein 61; RNA interference; Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2015.01.018

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To explore the inhibition effect of Cysteine-rich 61(CCN1;Cyr61) specific siRNA expression vector on RNV in a mouse model of oxygen-induced retinopathy (OIR). Methods One hundred and twenty healthy C57BL/6J mice were chosen and randomly divided into the experimental group and control group, with 60 mice in each group. The experimental group was intravitreously injected with CCN1siRNA recombinant plasmids. The control group was injected with vector plasmids. Adenosine diphosphate-ase stained retina flat-mounts was performed to assess the retinal vascular profiles, retinal section with HE staining was applied to count the number of new vascular cell nuclei and the protein and mRNA expression of CCN1 and vascular endothelial growth factor (VEGF) were detected by immunohistochemistry, Western blot and Real-time RT-PCR. Results Compared with control group, regular distributions, good branches and reduced density of retinal neovascularization were observed in the experimental group. The number of nucleus of vascular endothelial cells breaking through the inner limiting membrane was obviously less in the experimental group than that in the control group (t=8.756, P < 0.05). The expression of CCN1 and VEGF were obviously decreased in the experimental group compared with the control group (all P < 0.05). Conclusion The development of RNV of ROP can be markedly inhibited by RNA interference targeting CCN1, and CCN1siRNA may provide an effective method for preventing vascular proliferative retinopathy.

Citation： DiYu, ZhangYiou, ChenXiaolong. Cysteine-rich 61 siRNA reduces retinal neovascularization of mice. Chinese Journal of Ocular Fundus Diseases, 2015, 31(1): 72-76. doi: 10.3760/cma.j.issn.1005-1015.2015.01.018 Copy

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2.	Wu WC, Yeh PT, Chen SN, et al.Effects and complications of bevacizumab use in patients with retinopathy of prematurity:a multicenter study in Taiwan[J]. Ophthalmology, 2011, 118(1):176-183.
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5.	Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method[J]. Methods, 2001, 25(4):402-408.
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7.	Park K, Chen Y, Hu Y, et al. Nanoparticle-mediated expression of an angiogenic inhibitor ameliorates ischemia-induced retinal neovascularization and diabetes-induced retinal vascular leakage[J]. Diabetes, 2009, 58(8):1902-1913.
8.	Chen Y, Du XY.Functional properties and intracellular signaling of CCN1/Cyr61[J].J Cell Biochem, 2007, 100(6):1337-1345.
9.	Haque I, De A, Majumder M, et al.The matricellular protein CCN1/Cyr61 is a critical regulator of Sonic Hedgehog in pancreatic carcinogenesis[J].J Biol Chem, 2012, 287(46):38569-38579.
10.	Jandova J, Beyer TE, Meuillet EJ, et al.The matrix protein CCN1/CYR61 is required forα(V)β5-mediated cancer cell migration[J].Cell Biochem Funct, 2012, 30(8):687-695.
11.	Watari H, Xiong Y, Hassan MK, et al.Cyr61, a member of ccn (connective tissue growth factor/cysteinerich 61/nephroblastoma overexpressed) family, predicts survival of patients with endometrial cancer of endometrioid subtype[J].Gynecol Oncol, 2009, 112(1):229-234.
12.	Leask A.Sonic advance:CCN1 regulates sonic hedgehog in pancreatic cancer[J].J Cell Commun Signal, 2013, 7(1):61-62.
13.	Long QZ, Zhou M, Liu XG, et al. Interaction of CCN1 with alphavbeta3 integrin induces P-glycoprotein and confers vinblastine resistance in renal cell carcinoma cells[J]. Anticancer Drugs, 2013, 24(8):810-817.
14.	You JJ, Yang CH, Chen MS, et al. Cysteine-rich 61, a member of the CCN family, as a factor involved in the pathogenesis of proliferative diabetic retinopathy[J].Invest Ophthalmol Vis Sci, 2009, 50(7):3447-3455.
15.	Masuda I, Matsuo T, Yasuda T, et al.Gene transfer with liposomes to the intraocular tissues by different routes of administration[J].Invest Ophthalmol Vis Sci, 1996, 345(9):1914-1920.
16.	Masuda I, Matsuo T, Yasuda T, et al.Gene transfer with liposomes to the intraocular tissues by different routes of administration[J].Invest Ophthalmol Vis Sci, 1996, 37(9):1914-1920.
17.	You JJ, Yang CM, Chen MS, et al. Regulation of Cyr61/CCN1 expression by hypoxia through cooperation of c-Jun/AP-1 and HIF-1alpha in retinal vascular endothelial cells[J]. Exp Eye Res, 2010, 91(6):825-836.
18.	Hanrahan F, Humphries P, Campbell M. RNAi-mediated barrier modulation:synergies of the brain and eye[J]. Ther Deliv, 2010, 1(4):587-594.
19.	Roy S, Nasser S, Yee M, et al. A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats[J]. Mol Vis, 2011, 17:3166-3174.
20.	Pierce EA, Foley ED, Smith LE.Regulation of vascular endothelial growth factor by oxygen in a model of retinopathy of prematurity[J].Arch Ophthalmol, 1996, 114(10):1219-1228.
21.	Gerhardt H.VEGF and endothelial guidance in angiogenic sprouting[J]. Organogenesis, 2008, 4(4):241-246.

1. Heidary G, Vanderveen D, Smith LE.Retinopathy of prematurity:current concepts in molecular pathogenesis[J]. Semin Ophthalmol, 2009, 24(2):77-81.
2. Wu WC, Yeh PT, Chen SN, et al.Effects and complications of bevacizumab use in patients with retinopathy of prematurity:a multicenter study in Taiwan[J]. Ophthalmology, 2011, 118(1):176-183.
3. Koon HW, Shih DQ, Hing TC, et al.Substance P induces CCN1 expression via histone deacetylase activity in human colonic epithelial cells[J].Am J Pathol, 2011, 179(5):2315-2326.
4. Smith LE, Wesolowski E, Mclellan A, et al.Oxygen-induced retinopathy in the mouse[J].Invest Ophthalmol Vis Sci, 1994, 35(1):101-111.
5. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method[J]. Methods, 2001, 25(4):402-408.
6. Zhang Q, Zhang J, Guan Y, et al. Suppression of retinal neovascularization by the iNOS inhibitor aminoguanidine in mice of oxygen-induced retinopathy[J]. Graefe's Arch Clin Exp Ophthalmol, 2009, 247(7):919-927.
7. Park K, Chen Y, Hu Y, et al. Nanoparticle-mediated expression of an angiogenic inhibitor ameliorates ischemia-induced retinal neovascularization and diabetes-induced retinal vascular leakage[J]. Diabetes, 2009, 58(8):1902-1913.
8. Chen Y, Du XY.Functional properties and intracellular signaling of CCN1/Cyr61[J].J Cell Biochem, 2007, 100(6):1337-1345.
9. Haque I, De A, Majumder M, et al.The matricellular protein CCN1/Cyr61 is a critical regulator of Sonic Hedgehog in pancreatic carcinogenesis[J].J Biol Chem, 2012, 287(46):38569-38579.
10. Jandova J, Beyer TE, Meuillet EJ, et al.The matrix protein CCN1/CYR61 is required forα(V)β5-mediated cancer cell migration[J].Cell Biochem Funct, 2012, 30(8):687-695.
11. Watari H, Xiong Y, Hassan MK, et al.Cyr61, a member of ccn (connective tissue growth factor/cysteinerich 61/nephroblastoma overexpressed) family, predicts survival of patients with endometrial cancer of endometrioid subtype[J].Gynecol Oncol, 2009, 112(1):229-234.
12. Leask A.Sonic advance:CCN1 regulates sonic hedgehog in pancreatic cancer[J].J Cell Commun Signal, 2013, 7(1):61-62.
13. Long QZ, Zhou M, Liu XG, et al. Interaction of CCN1 with alphavbeta3 integrin induces P-glycoprotein and confers vinblastine resistance in renal cell carcinoma cells[J]. Anticancer Drugs, 2013, 24(8):810-817.
14. You JJ, Yang CH, Chen MS, et al. Cysteine-rich 61, a member of the CCN family, as a factor involved in the pathogenesis of proliferative diabetic retinopathy[J].Invest Ophthalmol Vis Sci, 2009, 50(7):3447-3455.
15. Masuda I, Matsuo T, Yasuda T, et al.Gene transfer with liposomes to the intraocular tissues by different routes of administration[J].Invest Ophthalmol Vis Sci, 1996, 345(9):1914-1920.
16. Masuda I, Matsuo T, Yasuda T, et al.Gene transfer with liposomes to the intraocular tissues by different routes of administration[J].Invest Ophthalmol Vis Sci, 1996, 37(9):1914-1920.
17. You JJ, Yang CM, Chen MS, et al. Regulation of Cyr61/CCN1 expression by hypoxia through cooperation of c-Jun/AP-1 and HIF-1alpha in retinal vascular endothelial cells[J]. Exp Eye Res, 2010, 91(6):825-836.
18. Hanrahan F, Humphries P, Campbell M. RNAi-mediated barrier modulation:synergies of the brain and eye[J]. Ther Deliv, 2010, 1(4):587-594.
19. Roy S, Nasser S, Yee M, et al. A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats[J]. Mol Vis, 2011, 17:3166-3174.
20. Pierce EA, Foley ED, Smith LE.Regulation of vascular endothelial growth factor by oxygen in a model of retinopathy of prematurity[J].Arch Ophthalmol, 1996, 114(10):1219-1228.
21. Gerhardt H.VEGF and endothelial guidance in angiogenic sprouting[J]. Organogenesis, 2008, 4(4):241-246.

Chinese Journal of Ocular Fundus Diseases

Cysteine-rich 61 siRNA reduces retinal neovascularization of mice

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