[Expression and Biological Function of SPOP in Acute Myeloid Leukemia].

Abstract

Objective: To study the expression of SPOP in patients with acute myeloid leukemia (AML) and its effect on proliferation, apoptosis and cycle of AML cells.

Methods: RT-qPCR was used to detect the expression of SPOP mRNA in bone marrow samples of patients with newly diagnosed AML and normal controls. The stable overexpression of SPOP in AML cell lines THP-1 and U937 were constructed by liposome transfection. The effect of SPOP on cell proliferation was detected by CCK-8, and the effect of SPOP on apoptosis and cell cycle was detected by flow cytometry. The expressions of anti-apoptotic protein Bcl-2 and apoptotic protein Bax, Caspase3 were detected by Western blot.

Results: The median expression level of SPOP mRNA in normal control group was 0.993 1(0.6303, 1.433), while that in AML group was 0.522 1(0.242 2, 0.723 7). The expression level of SPOP in AML group was significantly lower than that in normal control group ( P < 0.001). After the overexpression of SPOP, the proportion of apoptotic cells in the U937 overexpression group and THP-1 overexpression group was 10.9%±0.3% and 4.6%±015%, which were higher than 8.9%±0.3% and 3.0%±0.30% in the Empty Vector group, respectively (both P < 0.05). The expression of Caspase3 in U937 overexpression group and THP-1 overexpression group was 1.154±0.086 and 1.2±0.077, which were higher than 1 in Empty Vector group, respectively (both P < 0.05). The ratio of Bax/Bcl-2 in U937 overexpression group and THP-1 overexpression group was 1.328±0.057 and 1.669±0.15, which were higher than 1 in Empty Vector group, respectively (both P < 0.05). In the cell proliferation experiment, the number of cells in the U937 overexpression group and THP-1 overexpression group were both slightly lower than those in the Empty Vector group, but the differences were not statistically significant (P >0.05). In the cell cycle experiment, the proportion of G₁ cells in the U937 overexpression group and THP-1 overexpression group were both slightly higher than those in the Empty Vector group, but the differences were not statistically significant (P >0.05).

Conclusion: SPOP can promote the apoptosis of leukemic cells, and its mechanism may be related to down-regulation of Bcl-2 expression and up-regulation of Bax and Caspase3 expression.