Study on effect of vimentin glycosylated mutant plasmid on differentiation of PC12 cells
10.3969/j.issn.1671-8348.2024.08.001
- VernacularTitle:波形蛋白糖基化突变质粒对PC12细胞分化的影响研究
- Author:
Xiaoting SHAO
1
;
Yuxuan DAI
;
Lingjing YING
;
Meixian CHEN
Author Information
1. 浙江大学医学院附属金华医院/金华市中心医院儿科,浙江金华 321000
- Keywords:
glycosylation;
vimentin;
differentiation;
plasmid;
mutation
- From:
Chongqing Medicine
2024;53(8):1121-1125,1131
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a vimentin glycosylated mutant plasmid,and to study its impact on rat adrenal chromaffin cell (PC12) cell differentiation.Methods The vimentin glycosylated mutant plasmid was constructed,and the gel electrophoretic was performed and the sequencing identification was performed. The empty plasmid pcDNA3.1b (control group),vimentin plasmid (Vim group) and vimentin glycosylated mutant plasmid (Vim mut group) were used to transfect PC12 cells,which were cultured for 0,3 d in the con-dictions of treatment and non-treatment of exogenous oligosaccharides (Cyclo-ManN pro) respectively,and the cell differentiation records and quantitative analysis (the cell average neurite length and differentiated neu-rite cells percentage) were performed.Results A recombinant plasmid containing a glycosylation site muta-tion site 19(T-G),97(A-G),100(T-G) was successfully constructed (about 1404 bp in size).Before and after Cyclo-ManN pro treatment,the average neurite length of PC12 cells and the percentage of differentiated neu-rite cells in the Vim group were greater than those in the control group and the Vim mut group[(61.98±19.03)μm vs.(51.09±14.45)μm,(51.49±14.78)μm,(6.60±0.25)％ vs. (4.27±0.18)％,(4.76±0.33)％;(78.01±18.31)μm vs. (69.98±12.85)μm,(68.45±13.84)μm,(10.62±0.25)％ vs. (8.11±1.22)％,(5.89±0.60)％],the difference was statistically significant (P<0.05).Conclusion The glycosyla-tion modification of vimentin could be used for PC12 cell differentiation.
