The effect of PKD1 gene on autophagy in aortic smooth muscle cells_Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Authors：

YANG Guifang ¹ ,  CHAI Xiangping ¹ , PENG Wen ¹ , ZHOU Yang ¹ , SHENG Lijuan ²

1. Department of Emergency Medicine, The Second Xiangya Hospital, Central South University, Changsha, 410011, P.R.China;
2. Department of Neurology, The Second Xiangya Hospital, Central South University, Changsha, 410011, P.R.China;

Corresponding author：

CHAI Xiangping, Email: chaixiangping@csu.edu.cn

Keywords：

PKD1 gene; vascular smooth muscle cell; lentivirus; autophagy

DOI：

10.7507/1007-4848.201904029

Video：

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Objective To explore the effects of PKD1 gene on mouse aortic smooth muscle (MOVAS) cells autophagy.Methods The shRNA and over-expression lentiviral vectors for the target gene of PKD1 were constructed. MOVAS cells were infected by a number of successful packaging shRNA (PKD1 knockdown) or ETS-1 (PKD1 over-expressing) lentiviral vectors, and qPCR was used to test interference and over-expressing effects. Then qPCR and Western blotting were used to detect the expression levels of autophagy markers including Atg5, Beclin1 and LC3 in control group, shPKD1 group and ETS-1 group.Results Compared with the control group, PKD1 mRNA level was decreased in the shPKD1 group (P<0.05); ETS-1 and PKD1 mRNA levels were increased in the ETS-1 group (P<0.05). In contrast with the control group, the mRNA levels of autophagy markers including Atg5 (P<0.05) and Beclin1 (P<0.01) were obviously decreased in the shPKD1 group, but they were obviously increased in the ETS-1 group (P<0.001). Protein levels of Atg5, Beclin1 and LC3 were significantly decreased in the shPKD1 group (P<0.05), but they were increased obviously in the ETS-1 group (P<0.05) in contrast with the control group.Conclusion PKD1 gene is involved in MOVAS cells autophagy, low expression of PKD1 gene can inhibit autophagy and high expression of PKD1 promotes autophagy in vascular smooth muscle cells.

Citation： YANG Guifang, CHAI Xiangping, PENG Wen, ZHOU Yang, SHENG Lijuan. The effect of PKD1 gene on autophagy in aortic smooth muscle cells. Chinese Journal of Clinical Thoracic and Cardiovascular Surgery, 2020, 27(5): 569-573. doi: 10.7507/1007-4848.201904029 Copy

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1. Boya P, Reggiori F, Codogno P. Emerging regulation and functions of autophagy. Nat Cell Biol, 2013, 15(7): 713-720.
2. Hassane S, Claij N, Jodar M, et al. Pkd1-inactivation in vascular smooth muscle cells and adaptation to hypertension. Lab Invest, 2011, 91(1): 24-32.
3. Hassane S, Claij N, Lantinga-van Leeuwen IS, et al. Pathogenic sequence for dissecting aneurysm formation in a hypomorphic polycystic kidney disease 1 mouse model. Arterioscl Throm Vas, 2007, 27(10): 2177-2183.
4. Boulter C, Mulroy S, Webb S, et al. Cardiovascular, skeletal, and renal defects in mice with a targeted disruption of the Pkd1 gene. P Natl Acad Sci USA, 2001, 98(21): 12174-12179.
5. GuifangY, Sheng LJ, Peng W, et al. Comorbidity: A novel insight to aortic dissection. Int JCardiol, 2016, 207: 53-54.
6. Grantham JJ. Clinical practice. New Engl J Med, 2008, 359(14): 1477-1485.
7. Fukunaga N, Yuzaki M, Nasu M, et al. Dissecting aneurysm in a patient with autosomal dominant polycystic kidney disease. Ann Thorac Cardiovasc, 2012, 18(4): 375-378.
8. Tai S, Hu XQ, Peng DQ, et al. The roles of autophagy in vascular smooth muscle cells. Int JCardiol, 2016, 211: 1-6.
9. You Z, Qian H, Wang C, et al. Regulation of vascular smooth muscle cell autophagy by DNA nanotube-conjugated mTOR siRNA. Biomaterials, 2015, 67: 137-150.
10. Mizushima N, Komatsu M. Autophagy: Renovation of cells and tissues. Cell, 2011, 147(4): 728-741.
11. Levine B, Kroemer G. Autophagy in the pathogenesis of disease. Cell, 2008, 132(1): 27-42.
12. Schiattarella GG, Hill JA. Therapeutic targeting of autophagy in cardiovascular disease. J Mol Cell Cardiol, 2016, 95: 86-93.
13. Lambeth LS, Smith CA. Short hairpin RNA-mediated gene silencing. Method Mol Biol, 2013, 942: 205-232.
14. Rossi JJ. Expression strategies for short hairpin RNA interference triggers. Hum Gene Ther, 2008, 19(4): 313-317.
15. Puri S, Rodova M, Islam MR, et al. Ets factors regulate the polycystic kidney disease-1 promoter. Biochem Biophys Res Commun, 2006, 342(4): 1005-1013.
16. Lantinga-van Leeuwen IS, Leonhard WN, Dauwerse H, et al. Common regulatory elements in the polycystic kidney disease 1 and 2 promoter regions. Eur J Hum Genet, 2005, 13(5): 649-659.

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