Basic & Clinical Medicine ›› 2021, Vol. 41 ›› Issue (10): 1410-1416.

• Original Articles • Previous Articles     Next Articles

RNA-seq analysis on cisplatin down-regulating expression of DRG neurons' axon guidance related genes

XU Xian-lin, SONG Si-yuan, YU Chun-bo, LU Hong-ling, ZHUO Ming*   

  1. Department of Biochemistry and Molecular Biology, Zunyi Medical University,Zunyi 563099, China
  • Received:2020-12-14 Revised:2021-04-01 Published:2021-09-29
  • Contact: *ming_zhuo@hotmail.com

Abstract: Objective To explore the effect of cisplatin on the expression of axon guidance related gene in mouse dorsal root ganglion (DRG) neurons using transcriptome sequencing technique, and to further explore the neurotoxic of cisplatin. Methods The primary cultures of DRG neurons was isolated from Kunming mice and divided into control group and injury group. Neurons in the control group were cultured for 24 hours, and those in the injury group were pre-cultured for 4 hours and then treated with 20 μmol/L cisplatin for 20 hours. To observe the neurites and the growth cones in two groups by the NF200 immunofluorescence staining. Total RNA was extracted from the cells, and transcriptome sequencing was performed using RNA-seq technology. The axon guidance related genes were searched through literature and database (total 231), and the differential expression of related genes was analyzed. Real-time fluorescence quantitative PCR (qPCR) was used to verify the expression of related genes (Cxcl12,Ext1,Gdnf,Plxna4,Nrp2,Apbb2,Sema6a,Nrp1,Nrcam,Slit1,Ank3,Ncam1,Sema7a,Dag1,Klf7,Ntn1). Results Compared with the control group, the neurites of the injury group were significantly shortened and reduced, and the growthcones showed barb retraction. A large number of genes were down-regulated in the injury group as compared to the control group, among them, 49 genes related to axon guidance pathway were down-regulated. The proteins interacted closely and mainly focus on Ntn1,Nrp1,Nrp2,Sema6a,plxna2,plxna4,Slit2,Robo1.The qPCR verification of related genes expression was consistent with the trend of RNA-seq results. Conclusions The neurotoxicity mechanism of cisplatin may be related to the down-regulation of many axon guidance related genes.

Key words: cisplatin, dorsal root ganglion, axon guidance, transcritome sequencing, differential gene

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