胞质和核定位a-突触核蛋白过表达对人神经母细胞瘤细胞的影响*

南京大学学报(自然科学版) ›› 2013, Vol. 49 ›› Issue (1): 109–115.

胞质和核定位a-突触核蛋白过表达对人神经母细胞瘤细胞的影响*

夏娟1，高静1，熊御云2，邱晶1，马瑞3，钱进军4**

出版日期:2015-09-22 发布日期:2015-09-22
作者简介: (1.江苏大学药学院，镇江，212013;2.江苏大学基础医学与医学技术学院，镇江，212013;
3.江苏大学临床医学院，镇江，212013;.江苏大学附属第四医院，镇江，212013)
基金资助:
江苏省自然科学基金(BK2008249)，江苏省“六大人才高峰”资助项日(2007038)

Effects of cytoplasm localized or nuclear localized overexpression
α-synuclein on SH-SYSY cells

Xia Juan1,Gao Jing1 ,Xionn Yu-Yun２ ,Qiu Jing1 ,Ma Rui3 ,Qian Jin一Jun4

Online:2015-09-22 Published:2015-09-22
About author:(1 .School of Pharmacy,Jiangsu University,Zhenjiang,212013,China;
2. School of Medical Science and Laboratory Medicine,Jiangsu University,Zhenjiang,212013,China;
3. School of Medicine,Jiangsu University,Zhenjiang,212013,China;
4. Fourth Peoplc’s Hospital of Zhenjiang J一angsu University,在henjiang,212013,China)

摘要/Abstract

摘要： 探讨α一突触核蛋白在人神经母细胞瘤(SH-SY5Y)细胞中定位过表达的影响.研究应用
PLVU-SNCA-NES(NLS)-2A EGFP重组慢病毒载体感染SH-SY5Y细胞，通过观察绿色荧光蛋白(EG-
FP)检测目的基因在细胞中的表达情况.形态观察与MTT检测相结合，分析胞质和核定位α-突角虫核蛋
白过表达对SH-SY5Y细胞增殖的影响，同时测定鱼藤酮、H^₂O_²对胞质和核定位a突角虫核蛋白过表达
SH-SY5Y细胞活性的影响.MitoTrackcr染色法测定胞质和核定位a突角虫核蛋白过表达的SH-SY5Y
细胞中线粒体数目的变化.结果发现，α一突触核蛋白胞质定位过表达使得SH-SY5Y细胞培养96 h时活
性显著增强，24 h,48 h,96 h时细胞中线粒体的数目显著增加，增长比率分别为91.18%,41.15%,50.
21%;a一突触核蛋白核定位过表达的SH-SY5Y细胞相对正常细胞表现出较低的细胞活性，在培养48 h,
72 h后细胞存活率显著下降.400 nmol/L鱼藤酮损伤24h后，α-突触核蛋白核定位过表达的SH-SY5Y
细胞活性显著高于正常细胞角藤酮损伤组.50-800?mol/L H_²O_²处理条件下，α一突触核蛋白的定位过
表达对SH-SY5Y细胞活性的影响并不显著.研究结果表明α一突触核蛋白胞质定位过表达表现出一定
的神经保护作用，α一突触核蛋白核定位过表达也能使SH-SY5Y细胞的活性发生改变.

Abstract: To study the effects of overexpression α-synuclcin localized in cytoplasm or in nuclear on viability of SH-
SY5Y cells. The recombinant plasmid PLVU-SNCA-NES(NLS)-2A-EUFP was transfected into SH-SY5Y cells by
lentivirus vector.Then the expression of reporter gene EUFP is detected by fluorescence microscopy; MTT to
analyze the viability of SH-SY5Y,SH-SY5Y(NES)and SH-SY5Y(NLS)ce1ls;MitoTraker Red to detect the amount
of mitochondria. After transfected with the recombinant plasmid PLVU-SNCA-NESC NLS)-2A-EUFP by lentivirus
vector for 48 h,the expression of EUFP is detected. Both the stable genetic SH-SY5Y(NES)and SH-SY5Y(NLS)arc
established.The viability of SH-SY5Y(NES)cclls is significantly enhanced compared with normal control at 96 h，
and the amount of mitochondria in SH-SY5Y(NES)is significantly increased at 24 h,48 h and 96 hand the incrcas-
ing rate is 91.18%,41.15%and 50. 21%，respectively. While the viability of SH-SY5Y(NLS)is decreased following
the transfection,it is surprising to find that following the administration of X00 nmol/I. rotenone, the viability of SH-
SY5Y(NLS)is much higher than SH-SY5Y cells. But with the treatment of 50一800 pmol/L H(),there is no sig-
nificant difference in survival rate occurred among all kinds of cells, In summary, crsynulcin cytoplasm
overexpression may enhance the viability of SH-SY5Y cells, crsynuclcin nuclear localization overexpression can also
change the viability of SH-SY5Y cells.

夏娟1，高静1，熊御云2，邱晶1，马瑞3，钱进军4**. 胞质和核定位a-突触核蛋白过表达对人神经母细胞瘤细胞的影响*
[J]. 南京大学学报(自然科学版), 2013, 49(1): 109–115.

Xia Juan1,Gao Jing1 ,Xionn Yu-Yun２ ,Qiu Jing1 ,Ma Rui3 ,Qian Jin一Jun4. Effects of cytoplasm localized or nuclear localized overexpression
α-synuclein on SH-SYSY cells
[J]. Journal of Nanjing University(Natural Sciences), 2013, 49(1): 109–115.

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