Effects of ubiquitin-like modifier activating enzyme 1 on proliferation and apoptosis of acute myeloid leukemia cells and its molecular mechanism
10.13200/j.cnki.cjb.004470
- VernacularTitle:泛素样修饰物激活酶1对急性髓系白血病细胞增殖和凋亡的影响及其作用机制
- Author:
ZHANG Aili
- Publication Type:Journal Article
- Keywords:
Leukemia;
Ubiquitin-like modifier activating enzyme 1(UBA1);
HL60 cells;
THP-1 cells;
Cell proliferation;
Cell apoptosis
- From:
Chinese Journal of Biologicals
2025;38(04):399-406
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of ubiquitin-like modifier activating enzyme 1(UBA1) on the proliferation and apoptosis of acute myeloid leukemia(AML) cells(HL60 and THP-1 cells) and its mechanism, so as to evaluate the possibility of using UBA1 as a molecular marker and target for diagnosis and treatment of AML. Methods The expression of UBA1 in HL60 and THP-1 cells was inhibited via shRNA interference, and stable transfection cell lines were screened. The inhibition effect was detected by qPCR and Western blot. The effect of inhibiting UBA1 on proliferation of AML cells was measured by CCK-8 assay. Flow cytometry was used to detect the effect of inhibiting UBA1 on apoptosis of AML cells. The effects on the expression of apoptosis proteins(Bax, Bc12), cell cycle regulation related proteins(CDK4, CDK6 and CyclinD1) and MAPK signaling pathway related proteins(P-ERK, P-JNK, P-P38MAPK, T-ERK, T-P38) in AML cells were determined by Western blot. Results Following UBA1 knockdown, both HL60 and THP-1 cells exhibited a significant reduction in UBA1 mRNA transcription and protein expression levels(t = 2. 065-43. 591, each P < 0. 05). Cell proliferation capacity was significantly suppressed(t = 12. 274-17. 252, each P < 0. 05), while the apoptosis rate increased significantly(t = 12. 690-18. 855, P <0. 05). The pro-apoptotic protein Bax was significantly upregulated(t = 17. 094-20. 781, P < 0. 01), whereas the anti-apoptotic protein Bcl-2 was downregulated(t = 42. 494-53. 050, P < 0. 01). The expression levels of cell cycle regulatory proteins CDK4, CDK6, and Cyclin D1 all significantly decreased(t = 12. 193-51. 666, each P < 0. 05). The expression levels of P-ERK and P-P38MAPK in MAPK signaling pathway increased significantly(t = 3. 759-10. 822, each P < 0. 05),but the expression levels of P-JNK, T-ERK and T-P38 had no statistically significant difference(t = 0. 133-1. 794, each P >0. 05). Conclusion Interference with the expression of UBA1 can inhibit the proliferation of HL60 and THP-1 cells and promote their apoptosis, of which the mechanism may be related to the increased expression of P-ERK and P-P38MAPK proteins in MAPK signaling pathway.
