Intervene of Hedgehog signal on expression of proliferating cell nuclear antigen in renal tubular epithelial cells
1.Department of Laboratory Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015; 2.Wenzhou Key Laboratory of Surgery, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015
LU Hong1,WU Lianfeng1,LIN Chengcheng2, et al. Intervene of Hedgehog signal on expression of proliferating cell nuclear antigen in renal tubular epithelial cells[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2015, 45(1): 14-.
Abstract：Objective: To explore and investigate the activation and inhibition of Hedgehog signal regulated on expression of proliferating cell nuclear antigen (PCNA) in renal tubular epithelial cells (NRK-52E). Methods: NRK-52E cells were randomly divided into control group (treated with solvent only), activated group (treated with Shh at the concentrations with 10 μg/L) and treatment group (treated with Shh at the concentration of 10 μg/L plus cyclopamine at the concentrations with 5 μmol/L). After cultured for 24 h, immunofluorescence staining was used to evaluate the expressions of Ptch1, Smo, Gli1, and PCNA. The expression levels of Ptch1 and Smo mRNA were quantified by real-time RT-PCR. Results: NRK-52E cells without drug treatment maintain slowed proliferation state, and HH signal was not activated, and PCNA was in low expression. Treatment with exogenous recombinant protein Shh for 24 h, the mRNA or protein expression levels of Smo and Gli1 were significantly increased, and the expression of Ptch1 was decreased, suggesting that Shh activates the HH signal (P<0.05). In this process, the expression of PCNA protein was significantly increased. NRK-52E cells showed proliferation, and morphologic change occurred. After cyclopamine intervention, the activity of HH signal was decreased (P<0.05), and PCNA expression was also inhibited. In addition, abnormal proliferation of NRK-52E cells was inhibited, and increased apoptosis and necrosis cells. Conclusion: HH signal is involved in the proliferation of renal tubular epithelial cell. Regulating the activation of HH signal influences PCNA expression and cell proliferation.
[1] Yauch RL, Gould SE, Scales SJ, et al. A paracrine requirement for hedgehog signalling in cancer[J]. Nature, 2008, 455(7211): 406-410.
[2] Onishi H, Katano M. Hedgehog signaling pathway as a new therapeutic target in pancreatic cancer[J]. World J Gastroenterol, 2014, 20(9): 2335-2342.
[3] Omenetti A, Porrello A, Jung Y, et al. Hedgehog signaling regulates epithelial-mesenchymal transition during biliary fibrosis in rodents and humans[J]. J Clin Invest, 2008, 118 (10)3331-3342.
[4] 白永恒, 洪炜龙, 刘彪, 等. Sonic Hedgehog信号参与马兜铃酸损伤肾小管上皮细胞的体外实验研究[J]. 中华肾脏病杂志, 2013, 29(4): 288-292.
[5] 白永恒, 陆红, 周琴, 等. Sonic Hedgehog信号通路在单侧输尿管梗大鼠肾组织中的表达变化及意义[J]. 中国病理生理杂志, 2012, 28(12): 2227-2232.
[6] Katoh Y, Katoh M. Hedgehog signaling pathway and gastrointestinal stem cell signaling network[J]. Int J Mol Med, 2006, 18(6): 1019-1023.
[7] Ding H, Zhou D, Hao S, et al. Sonic hedgehog signaling mediates epithelial-mesenchymal communication and promotes renal fibrosis[J]. J Am Soc Nephrol, 2012, 23(5): 801-813.
[8] Mill P, Mo R, Fu H, et al. Sonic hedgehog-dependent activation of Gli2 is essential for embryonic hair follicle Development[J]. Genes Dev, 2003, 17(2): 282-294.
[9] 胡丽萍, 陆红, 白永恒, 等. 小G蛋白Rac1在马兜铃酸诱导肾小管上皮细胞损伤中的作用及意义[J]. 温州医学院学报, 2013, 43(12): 775-778.
[10] Tremblay MR, Nevalainen M, Nair SJ, et al. Semisynthetic cyclopamine analogues as potent and orally bioavailable hedgehog pathway antagonists[J]. J Med Chem, 2008, 51 (21): 6646-6649.
