Effect of silencing KAT7 gene on the production of MCP⁃1 in rat glomerular messangial cells induced by sublytic C5b⁃9 stimulation
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    Abstract:

    Objective:To explore the effect of silencing rat(lysine acetyltransferase 7) KAT7 gene on the production of monocyte chemotactic protein-1(MCP-1) in rat glomerular messangial cells(GMC) by sublytic C5b-9 stimulation. Methods:Four kinds of short hairpin RNA(shRNA) stargeting KAT7 gene were synthesized and cloned into eukaryotic expression vector pGCsi-U6/Neo/GFP/shRNA. The recombinant plasmids were transfected into cultured rat GMC by NeonTM transfection system,and KAT7 protein in the transfected cells was detected by Western blot to find out the optimal shRNA against KAT7 gene. Moreover,the expression of MCP-1 both in GMC and in the supernatant was measured by real-time PCR and ELISA assays. Results:Nucleotide sequencing demonstrated that the constructed KAT7 shRNAs were correct. Western blot experiment showed that the shKAT7-2 could effectively silence the target gene. Meanwhile,after the knockdown of KAT7 by shRNA in the GMC,the production of MCP-1 was significantly decreased upon sublytic C5b-9 stimulation. Conclusion:The rat eukaryotic expression vector shKAT7 was successfully constructed. It is preliminarily confirmed that the expression of KAT7 could obviously promote the production of MCP-1 in rat GMC treated by sublytic C5b-9.

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宫雅娟,张 婧,刘 玉,赵 聃,邱 文,夏 露,王迎伟.沉默大鼠KAT7 基因对亚溶解型 C5b⁃9 刺激 GMC 诱导趋化因子 MCP⁃1 生成的影响[J].南京医科大学学报(自然科学版英文版),2018,(4):423-428.

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  • Received:September 15,2017
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  • Online: April 26,2018
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