Basic & Clinical Medicine ›› 2017, Vol. 37 ›› Issue (3): 369-375.

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Construction of BCR-ABL SH3-T79Y mutant recombinant adenovirus vectors and its promotion on apoptosis of K562/G01 cells


  • Received:2016-01-14 Revised:2016-07-07 Online:2017-03-05 Published:2017-02-23

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 SH3-T79Y mutant was obtained. 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 phosphorylation was detected by Western-blot. Results: The vectors were successfully constructed. Transfection efficiency was more than 80% after transferring into K562/G01 cells for 72h; 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). Conclusion: Successfully constructed the SH3-T79Y mutant recombinant adenovirus vectors and proved it could promote the apoptosis of K562/G01 cells by inhibiting BCR-ABL and CrkL phosphorylation.

Key words: chronic myeloid leukemia, BCR-ABL SH3 mutant, recombinant adenovirus, K562/G01 cells