Effect of siRNA-interfered endogenous Kit6.2 gene in PC12 cells on rotenone-induced cell damage and signal pathway
10.3969/j.issn.1009-0126.2017.10.020
- VernacularTitle:小干扰RNA干扰PC12细胞内源性Kir6.2基因后对鱼藤酮诱导的细胞损伤及信号通路研究
- Author:
kun Xiao LIU
1
;
Gang WANG
;
Xu CHEN
Author Information
1. 200235,上海市第八人民医院神经内科
- Keywords:
RNA,small interfering;
PC12 cells;
Parkinson disease;
rotenone;
gene expression
- From:
Chinese Journal of Geriatric Heart Brain and Vessel Diseases
2017;19(10):1085-1089
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effect of siRNA-interfered endogenous Kir6.2 gene deletion on rotenone-induced cell damage and signal pathway according to the expression of siRNA-interfered endogenous Kir6.2 gene.Methods The expression of endogenous Kir6.2 gene in PC12 cells was detected by RT-PCR and Western bolt.The viability of PC12 cells was tested using WST-1.The expression of PKC and phosphorylated PKC in PC12 cells was detected by Western blot in negative control group,siRNA/Kir6.2 group,siRNA/Kir6.2 + rotenone group,siRNA/Kir6.2+ rotenone+PKC inhibitor group before and afer treatment with rotenone.Results RT-PCR showed that the interference was successful in siRNA/Kir6.2/group 1 and siRNA/Kir6.2/ group 2.Western blot displayed that the expression level of endogenous Kir 6.2 gene was significantly lower in siRNA/Kir6.2/ group 1 than in negative control group (0.55±0.07 vs 0.89±0.09,P＜ 0.05).WST-1 revealed that the viability of PC12 cells was significantly lower in siRNA/Kir6.2 group than in negative control group (P＜0.05).Western blot demonstrated that the expression of PKC and phosphorylated PKC was significantly lower in siRNA/Kir6.2+-rotenone+PKC inhibitor group than in the other three group (P＜0.05).Conclusion Endogenous Kir6.2 can protect PC12 cells against the toxicity of rotenone and plays an important role in regulating the rotenone-induced viability of PC12 cells by activating the phosphorylated PKC.
