[1]周玉帅,殷竞争,张瑞锋,等.转染TDP25对SHSY5Y细胞自噬蛋白表达及细胞活力、凋亡的影响*[J].郑州大学学报(医学版),2015,(03):393-396.
 ZHOU Yushuai,YIN Jingzheng,ZHANG Ruifeng,et al.Expression of autophagy proteins and cell vitality,apoptosis of SHSY5Y cells transfected with TDP25[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2015,(03):393-396.
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转染TDP25对SHSY5Y细胞自噬蛋白表达及细胞活力、凋亡的影响*()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2015年03期
页码:
393-396
栏目:
应用研究
出版日期:
2015-05-20

文章信息/Info

Title:
Expression of autophagy proteins and cell vitality,apoptosis of SHSY5Y cells transfected with TDP25
作者:
周玉帅12殷竞争12张瑞锋12滕军放12
1郑州大学第一附属医院神经内科 郑州 4500522河南省高等学校临床医学重点学科开放实验室 郑州 450052
Author(s):
ZHOU Yushuai12YIN Jingzheng12 ZHANG Ruifeng12 TENG Junfang12
1Department of Neurology, the First Affiliated Hospital,Zhengzhou University, Zhengzhou 450052 2KeyDisciplines Laboratory for Clinical Medicine of Henan Province,Zhengzhou 450052
关键词:
TDP25自噬细胞活力细胞凋亡
Keywords:
TDP25autophagycell vitalitycell apoptosis
分类号:
R742
文献标志码:
A
摘要:
目的:探讨TDP25对SHSY5Y细胞自噬蛋白表达及细胞活力、凋亡的影响。方法:将神经母细胞瘤细胞SHSY5Y分为3组,分别转染质粒GFPN1、GFPTDP43和GFPTDP25,24 h后应用免疫荧光法检测包涵体,Western blot法检测LC3、 Beclin1蛋白的表达情况。将SHSY5Y细胞转染质粒GFPTDP25后分为2组,一组加自噬抑制剂3MA,一组不加3MA,MTT比色法检测细胞活力,AnnexinVFITC/PI双染法检测细胞凋亡情况。结果:GFPN1组细胞胞质、胞核均见较强绿色荧光信号;GFPTDP43组较强的绿色荧光信号分布于胞核中,未见明显的包涵体形成;GFPTDP25组可见较强的绿色荧光信号颗粒状分布于胞质中,形成包涵体;3组Beclin1表达水平及LC3Ⅱ/LC3Ⅰ差异有统计学意义(F=192.700、247.420,P<0.05),GFPTDP25组Beclin1表达水平及LC3Ⅱ/LC3Ⅰ大于GFPTDP43组(P<0.05),后者大于GFPN1组(P<0.05)。加入3MA后,转染GFPTDP25的细胞细胞活力降低,细胞凋亡率增高(t=38.595、5.920,P<0.001)。结论:TDP25可能通过上调细胞巨自噬水平减弱其对细胞的损害。调控巨自噬水平对肌萎缩性侧索硬化症和额颞叶变性患者的病情进展可能起到一定的延缓作用。
Abstract:
To investigate the effect of TDP25 transfection on autophagy,vitality,and apoptosis of SHSY5Y cells.Methods: The recombinated plasmids GFPN1,GFPTDP43 and GFPTDP25 were transfected into SHSY5Y cells through eukaryotic cell transfection technique for 24 h,then immunofluorescence assay was performed to observe the form of ubiquitinated inclusions(UBIs), and the expressions of LC3 and Beclin1 were detected using Western blot assay. SHSY5Y cells were transfected with GFPTDP25, and treated with 3MA,then the cell vitality was measured using MTT, and apoptosis was assayed with flow cytometry. Results:UBIs were localized at the cytoplasm in the cells of GFPTDP25 group.There were significant differences in the expression levels of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰamong GFPN1,GFPTDP43 and GFPTDP25 groups(F=192.700,247.420,P<0.05), and the expression level of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰ in GFPTDP25 group was the highest(P<0.05). Cell vitality of the cells transfected with GFPTDP25 and treated with 3MA was lower and apoptosis rate were higher than cells only transfected with GFPTDP25(t=38.595,5.920,P<0.001).Conclusion: TDP25 might weaken the cell damage through upregulating the level of macroautophagy.It maybe helps to inhibit the progress of amyotrophic lateral sclerosis and frontotemporal lobar degeneration through a proper regulation of macroautophagy.

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备注/Memo

备注/Memo:
通信作者,男,1960年11月生,博士,教授,主任医师,研究方向:神经系统退行性疾病,Email:13838210077@163.com
更新日期/Last Update: 1900-01-01