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Archive > Volume 26 Issue 7 > 2019,26(7):736-742. DOI:10.3872/j.issn.1007-385X.2019.07.003 Prev Next
Basic Research
GADD45g exerts anti-tumor effect in acute myeloid leukemia via inhibition of E2F1
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Abstract:
[Abstract] Objective: To investigate the correlation between the expression of E2F1 and growth arrest and DNA damage inducible protein 45g (GADD45g) in patients with acute myeloid leukemia (AML), and to explore whether GADD45g exerts its induction of DNA damage, cell apoptosis, senescence, cell cycle arrest and drug sensitivity in AML through inhibition of E2F1. Methods: A total of 32 cases of bone marrow specimens from patients initially diagnosed as AML in Hospital of Blood Diseases Affiliated to Chinese Academy of Medical Sciences from January 2013 to December 2016, were selected for this study; In addition, AML cell lines (U937, HL60,THP-1, Molm-13) were also collected for this study. The mRNA expression of GADD45g and E2F1 in above mentioned specimens and cell lines by qPCR, and their correlation was also analyzed. The lentiviral vector over-expressing E2F1 (pLV-E2F1-RFP) was constructed to prepare recombinant lentivirus, which was then transfected Molm-13 and THP-1 cells that over-expressing GADD45g. Whether GADD45g exerts tumor inhibition effect on AML cells through inhibition of E2F1 was determined by comet assay, Annexin V/7AAD flow cytometry, β-galactosidase staining and PI staining flow cytometry etc. Results: The mRNA expression of GADD45g was negatively correlated with E2F1 in bone marrow of AML patients and AML cell lines (r=–0.663, P<0.01). Over-expression of GADD45g significantly inhibited the expression of E2F1 in AML cell lines (all P<0.01). Molm-13 and THP-1 cells that simultaneously over-expressing GADD45g and E2F1 were successfully constructed. Compared with the control group, the protein expressions of GADD45g and E2F1 in over-expression groups were significantly increased (all P<0.01). Compared with cells over-expressing GADD45g alone,simultaneous over-expression of both GADD45g and E2F1 significantly reduced the apoptosis, senescence and DNA damage (all P<0.01), and rescued cell cycle arrest in Molm-13 and THP-1 cells (all P<0.01), thus further reduced the chemo-sensitivity of AML cells caused by GADD45g over-expression (all P<0.01). Conclusion: GADD45g exerts anti-tumor effect in AML via inhibition of E2F1.
Keywords:
transcription factor E2F1 ; GADD45g ; acute myeloidleukemia ; cell apoptosis
Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81670158, No. 81470278, No. 81600138, No. 81700106), and the Tianjin Municipal Science and Technology Commission Grant (No. 17JCZDJC35100, No.15JCQNJC10300，No.17JCQNJC10800)
