关键词: 慢性粒细胞白血病, BCR-ABL SH3突变体, 重组腺病毒, K562/G01细胞

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