Construction of BCR-ABL SH3-T79Y mutant recombinant adenovirus vectors and its promotion on apoptosis of K 562/G01 cells
- VernacularTitle:BCR-ABL SH3-T79Y突变体 重组腺病毒载体的构建及促进白血病K562/G01细胞凋亡
- Author:
Liangxue WEN
;
Xin LIU
;
Hui LI
;
Ningshu HUANG
;
Zhenglan HUANG
;
Wenli FENG
- Keywords:
chronic myeloid leukemia;
BCR-ABL SH3 mutant;
recombinant adenovirus;
K562/G01 cells
- From:
Basic & Clinical Medicine
2017;37(3):369-375
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct BCR-ABL SH3-T79Y mutant recombinant adenovirus vectors and investigate its effects on apoptosis of K562/G01 cells.Methods SH3-T79Y mutant was amplified by overlapping PCR with pMig210 as template and cloned into recombinant adenovirus vectors .After identifying , packaging and amplifying , the recombinant adenovirus vectors containing SH 3-T79Y mutant was collected .Recombinant adenovirus vectors were transferred into K562/G01 cells.Then transfection efficiency was determinated , changes of cell morphology were observed by Wright 's staining , cell apoptosis was evaluated by flow cytometry , BCR-ABL and CrkL phospho-rylation was detected by Western blot .Results The vectors were successfully constructed .Transfection efficiency was more than 80%after transferring into K562/G01 cells for 72 h;there was obvious apoptosis phenomenon , cell apoptosis significantly increased to 32.46% compared with the control groups ( P<0.05 ) , BCR-ABL and CrkL phosphorylation significantly decreased and so did the expression of BCR-ABL( P<0.05 ) .Conclusions Success-fully constructed the SH 3-T79 Y mutant recombinant adenovirus vectors and it may promote the apoptosis of K 562/G01 cells by inhibiting BCR-ABL and CrkL phosphorylation .
