Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi · 2015
[The expression and role of the transcription factor C/EBPα in chronic myeloid leukemia].
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Zhang Guili, Dong Fei, Luan Caifu, Zhang Xia, Shao Huiyuan, Liu Jie, Sun Chengming
Аннотация
OBJECTIVE: To investigate the expression and the possible mechanism of the transcription factor C/EBPα in chronic myeloid leukemia（CML).
METHODS: Bone marrow samples from 50 CML patients（including 33 patients in chronic phase, 7 in accelerated phase and 10 in blast crisis）and peripheral blood specimens of 20 healthy donors were collected. The expression of C/EBPα gene and the effect of Imatinib on its expression was detected by RT- PCR. C/EBPα gene was inserted into lentivirus expression vector pLVX- EGFP- 3FLAG- Puro by recombinant DNA technology to construct C/EBPα stable expression in K562 cells. Cell proliferation was assayed by CCK-8. The expressions of Foxo3a and Bim genes were detected by RT-PCR.
RESULTS: The level of C/EBPα expression was significantly declined in CML patients compared with that of normal control group（P<0.01）and had negative correlation with bcr- abl expression（Spearman r=－ 0.505, P<0.01). The stable K562- C/EBPα cell line was successfully established and confirmed by RT-PCR and Western blot. Cell proliferation ability was lower in the K562- C/EBPα group than that in the non- transfection and mock-vehicle groups. The expressions of Foxo3a and Bim genes were 1.06 ± 0.06 and 0.53 ± 0.07, respectively, which was higher than that of nontransfection and mock-vehicle groups（P<0.01, P<0.05).
CONCLUSION: C/EBPα expression was decreased in CML patients, overexpression of C/EBPα could inhibit K562 cell growth.
METHODS: Bone marrow samples from 50 CML patients（including 33 patients in chronic phase, 7 in accelerated phase and 10 in blast crisis）and peripheral blood specimens of 20 healthy donors were collected. The expression of C/EBPα gene and the effect of Imatinib on its expression was detected by RT- PCR. C/EBPα gene was inserted into lentivirus expression vector pLVX- EGFP- 3FLAG- Puro by recombinant DNA technology to construct C/EBPα stable expression in K562 cells. Cell proliferation was assayed by CCK-8. The expressions of Foxo3a and Bim genes were detected by RT-PCR.
RESULTS: The level of C/EBPα expression was significantly declined in CML patients compared with that of normal control group（P<0.01）and had negative correlation with bcr- abl expression（Spearman r=－ 0.505, P<0.01). The stable K562- C/EBPα cell line was successfully established and confirmed by RT-PCR and Western blot. Cell proliferation ability was lower in the K562- C/EBPα group than that in the non- transfection and mock-vehicle groups. The expressions of Foxo3a and Bim genes were 1.06 ± 0.06 and 0.53 ± 0.07, respectively, which was higher than that of nontransfection and mock-vehicle groups（P<0.01, P<0.05).
CONCLUSION: C/EBPα expression was decreased in CML patients, overexpression of C/EBPα could inhibit K562 cell growth.