Effect of RNA Interference for c-Jun Gene on Proliferation of Rat Vascular Smooth Muscle Cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

ZHANG Hongwei ¹ , ZHOU Yu ² , HU Xinhua ³ , ZHANG Qiang ³

1.Department of General Surgery， The First Affiliated Hospital, Jiamusi University, Jiamusi 154001， Heilongjiang Province, China;;
2.Department of Tumour Surgery， The First Affiliated Hospital, Jiamusi University, Jiamusi 154001， Heilongjiang Province, China;;
3.Department of Vascular Surgery， The First Affiliated Hospital, China Medical University, Shenyang 110001， ChinaCorresponding Author: HU Xin-hua， E-mail： zhwwjl@126.com;

Corresponding author：

Keywords：

c-Jun gene; RNA interference; Vascular smooth muscle cell; Cell proliferation; Intimal hyperplasia; Activator protein-1

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Objective 　To investigate the influence of RNA interference targeting c-Jun gene on the proliferation of rat vascular smooth muscle cells (VSMCs).
Methods 　The experiment was performed with c-Jun siRNA （c-Jun siRNA group）, control reverse sequence siRNA (control siRNA group) or no siRNA (control group). VSMCs were transfected with siRNA targeting c-Jun gene by liposome. Effects of c-Jun siRNA on mRNA and protein expressions of c-Jun were examined by RT-PCR analysis and Western blot respectively. MTT test and 3H-TdR incorporation were used to detect VSMCs proliferation. Cell cycle analysis of VSMCs in vitro was determined by flow cytometer.
Results 　The expression levels of mRNA and protein of c-Jun in c-Jun siRNA group were significantly lower than those in control group （P＜0.05， P＜0.01). There was no significant difference between control group and control siRNA group （P gt;0.05）. Proliferation activity of VSMCs decreased significantly in c-Jun siRNA group compared with that in control group （P＜0.05） and VSMCs was blocked in the G0/G1 phase of cell cycle significantly （P＜0.05）. There was no significant difference between control group and control siRNA group （P gt;0.05）.
Conclusion 　c-Jun gene silenced by RNA interference can inhibit VSMCs proliferation effectively in vitro.

Citation： ZHANG Hongwei,ZHOU Yu,HU Xinhua,ZHANG Qiang. Effect of RNA Interference for c-Jun Gene on Proliferation of Rat Vascular Smooth Muscle Cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2009, 16(9): 723-728. doi: Copy

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13.	高正山，王树祥，李慧芳．综合干预疗法冠状动脉成形术加支架患者的远期疗效 [Ｊ]. 中国临床康复， 2002; 6(9)： 1257-1258.
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15.	Hiroi N, Nestler EJ. Nuclear memory: gene transcription and behavior [Ｊ]. Adv Pharmacol, 1998; 42: 1037-1041.
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17.	Faries PL, Marin ML, Veith FJ, et al. Immunolocalization and temporal distribution of cytokine expression during the development of vein graft intimal hyperplasia in an experimental model [Ｊ]. J Vasc Surg, 1996; 24(3): 463-471.
18.	何庆南，易著文. 转录因子AP-1的研究进展 [Ｊ]. 国外医学儿科学分册， 1999； 26（3）： 152-155.
19.	Caplen NJ, Parrish S, Imani F, et al. Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems [Ｊ]. Proc Natl Acad Sci USA, 2001; 98(17): 9742-9747.
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22.	Angel P, Karin M. The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation [Ｊ]. Biochim Biophys Acta, 1991; 1072(2-3): 129-157.
23.	周敬,　刁清,　王虹. 激活剂蛋白-1诱骗性寡核苷酸对大鼠颈总动脉损伤后新生内膜的影响 [Ｊ]. 疑难病杂志, 2006； 5(3): 164-166.
24.	Kume M, Komori K, Matsumoto T, et al. Administration of a decoy against the activator protein-1 binding site suppresses neointimal thickening in rabbit balloon-injured arteries [Ｊ]. Circulation, 2002; 105(10): 1226-1232.
25.	Gong H, Liu CM, Liu DP, et al. The role of small RNAs in human diseases: potential troublemaker and therapeutic tools [Ｊ]. Med Res Rev, 2005; 25(3): 361-381.

1. Davies MG, Hagen PO. Pathophysiology of vein graft failure: a review [Ｊ]. Eur J Vasc Endovasc Surg, 1995; 9(1): 7-18.
2. Ip JH, Fuster V, Badimon L, et al. Syndromes of accelerated atherosclerosis: role of vascular injury and smooth muscle cell proliferation [Ｊ]. J Am Coll Cardiol, 1990; 15(7): 1667-1687.
3. Bulkley BH, Hutchins GM. Accelerated “atherosclerosis”. A morphologic study of 97 saphenous vein coronary artery bypass grafts [Ｊ]. Circulation, 1977; 55(1): 163-169.
4. 郑江华, 刘玉娟,　时德. 血管再狭窄的分子生物学研究进展 [Ｊ]. 中国普外基础与临床杂志, 2004; 11（4）： 318-321.
5. 张震, 惠汝太. 转录因子AP-1与心血管疾病 [Ｊ]. 中国分子心脏病学杂志, 2003; 3（4）: 198-201.
6. Fire A, Xu S, Montgomery MK, et al．Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans [Ｊ]． Nature, 1998; 391(6669): 806-811.
7. Bernstein E, Caudy AA, Hammond SM, et al. Role for a bidentate ribonuclease in the initiation step of RNA interference [Ｊ]. Nature, 2001; 409(6818): 363-366.
8. 周晓莉, 雷寒, 柳青. 小鼠主动脉平滑肌细胞的培养 [Ｊ]. 基础医学与临床, 2005; 25(6): 564-567.
9. Campbell JH, Campbell GR．Culture techniques and their applications to studies of vascular smooth muscle [Ｊ]. Clin Sci (Lond), 1993; 85(5): 501-513.
10. Biswas N, Sanchez V, Spector DH. Human cytomegalovirus infection leads to accumulation of geminin and inhibition of the licensing of cellular DNA replication [Ｊ]. J Virol, 2003; 77(4): 2369-2376.
11. 蒋米尔, 赵海光. 血管外科实验研究的进展与发展方向 [Ｊ]. 中华实验外科杂志, 2005; 22(6): 648-650.
12. 崔广晖, 廖崇先，陈以旺. 经皮冠状动脉腔内成形术后再狭窄基因治疗现状与进展 [Ｊ]. 国外医学内科学分册, 2005; 32(10): 419-421.
13. 高正山，王树祥，李慧芳．综合干预疗法冠状动脉成形术加支架患者的远期疗效 [Ｊ]. 中国临床康复， 2002; 6(9)： 1257-1258.
14. Adcock IM. Transcription factors as activators of gene transcription: AP-1 and NF-kappa B [Ｊ]. Monaldi Arch Chest Dis, 1997; 52(2): 178-186.
15. Hiroi N, Nestler EJ. Nuclear memory: gene transcription and behavior [Ｊ]. Adv Pharmacol, 1998; 42: 1037-1041.
16. Chinenov Y, Kerppola TK. Close encounters of many kinds: Fos-Jun interactions that mediate transcription regulatory specificity [Ｊ]. Oncogene, 2001; 20(19): 2438-2452.
17. Faries PL, Marin ML, Veith FJ, et al. Immunolocalization and temporal distribution of cytokine expression during the development of vein graft intimal hyperplasia in an experimental model [Ｊ]. J Vasc Surg, 1996; 24(3): 463-471.
18. 何庆南，易著文. 转录因子AP-1的研究进展 [Ｊ]. 国外医学儿科学分册， 1999； 26（3）： 152-155.
19. Caplen NJ, Parrish S, Imani F, et al. Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems [Ｊ]. Proc Natl Acad Sci USA, 2001; 98(17): 9742-9747.
20. Elbashir SM, Harborth J, Lendeckel W, et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells [Ｊ]. Nature, 2001; 411(6836): 494-498.
21. 韩丽娜，顾瑛，刘凡光，等. c-jun反义核酸对自体移植静脉内膜增生和ET-1水平的影响 [Ｊ]. 放射免疫学杂志， 2004；17(6)： 429-431.
22. Angel P, Karin M. The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation [Ｊ]. Biochim Biophys Acta, 1991; 1072(2-3): 129-157.
23. 周敬,　刁清,　王虹. 激活剂蛋白-1诱骗性寡核苷酸对大鼠颈总动脉损伤后新生内膜的影响 [Ｊ]. 疑难病杂志, 2006； 5(3): 164-166.
24. Kume M, Komori K, Matsumoto T, et al. Administration of a decoy against the activator protein-1 binding site suppresses neointimal thickening in rabbit balloon-injured arteries [Ｊ]. Circulation, 2002; 105(10): 1226-1232.
25. Gong H, Liu CM, Liu DP, et al. The role of small RNAs in human diseases: potential troublemaker and therapeutic tools [Ｊ]. Med Res Rev, 2005; 25(3): 361-381.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect of RNA Interference for c-Jun Gene on Proliferation of Rat Vascular Smooth Muscle Cells

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