In vitro establishment of the overexpressing rat c-lun N-terminal Kinase model in SHSYSY
10.3760/cma.j.issn.1671-8925.2008.07.006
- VernacularTitle:大鼠JNK3超表达细胞模型的建立及特性研究
- Author:
Fang LI
1
;
Yun ZHANG
;
Yu-He YUAN
;
Jin-Feng HU
;
Nai-Hong CHEN
Author Information
1. 中国科学院理化技术研究所
- Keywords:
C-Jun N-terminal kinase 3;
Stably transfected
- From:
Chinese Journal of Neuromedicine
2008;7(7):670-673
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a PD model of overexpressing rat c-Jun N-terminal kinase 3 (JNK3) in SHSYSY(SHSYSY-rJNK3), and study the relationship between JNK3 and occuring of PD and AD. Methods Extract purificat RNA and get Rat JNK3 through RT-PCR. The pcDNA3. L/hisC-rJNK3 vector was established and stably transfected into SHSY5Y overexpressing rat JNK3. SHSYSY-rJNK3 was selected by Western blotting analysis. The growth rate and the sensitivity to MPI+ of SHSY5Y-rJNK3 were further evaluated by morphological observation, immunofluorescence and MTT assay. Results Successfully established SHSY5Y-rJNK3 series by transfeeted pcDNA3.1/hisC-rJNK3 vector in SHSY5Y over expressing rJNK3. There were morphological differences between SHSY5Y and SHSY5Y-rJNK3. The result of MTT showed that there were little differences between growth rate of both. Stimulated by MPP+ of different concentrations, the SHSY5Y-rJNK3 made more morphological changes in 100 um MPP+ than SHSYSY, and the results of MTT also demonstrated that SHSY5Y-rJNK3 was more sensitive to MPP+ compared to SHSYSY with lower cell viability. Conclusions Overexpressing JNK3 in SHSY5Y makes morphological changes on cells, also makes cells more sensitive to the poisons. Therefore, studying on SHSYSY-rJNK3 which is overexpressed JNK3 can reflect phenomenons and substance of apotosis which related to JNK3 more objective and accurately.
