[TMSB4X Promotes Proliferation and Metastasis of T-cell Acute Lymphoblastic Leukemia Cells by Regulating the...

To investigate the expression of TMSB4X in T-cell acute lymphoblastic leukemia (T-ALL) cells, as well as its effects on T-ALL cell proliferation and metastasis and the underlying mechanisms.

qRT-PCR and Western blot were used to detect the expression of TMSB4X mRNA and protein in human CD3⁺T cells from healthy donors and the human T-ALL cell lines Jurkat, MOLT-4, and CCRF-CEM. Jurkat cells with TMSB4X knockdown (TMSB4X-KD group) and CCRF-CEM cells with TMSB4X overexpressing (TMSB4X-OE group) were constructed via lentivirus infection, with corresponding control groups set up for each; additionally, Jurkat cells in the TMSB4X-KD group were co-incubated with 20 μmol/L SKL2001 (a Wnt/β-catenin signaling pathway agonist) for 24 hours ( TMSB4X-KD+SKL2001 group), while CCRF-CEM cells in the TMSB4X-OE group were co-incubated with 10 μmol/L MSAB (a Wnt/β-catenin signaling pathway inhibitor) for 24 hours (TMSB4X-OE+MSAB group). CCK8 assay, flow cytometry and Transwell assay were used to detect the cell proliferation, apoptosis, and migration ability, respectively, while Western blot was used to detect the expression of related proteins in each group.

The expression levels of TMSB4X mRNA and protein in the T-ALL cell lines (Jurkat, MOLT-4 and CCRF-CEM) were all significantly higher than those in CD3⁺T cells from healthy donors (P <0.01). Compared with the control group, Jurkat cells in the TMSB4X-KD group showed significantly decreased proliferation and migratory abilities (P <0.001), a significantly elevated apoptosis rate (P <0.001), and significantly downregulated expression of Wnt4 and β-catenin proteins (P <0.001); in contrast, CCRF-CEM cells in the TMSB4X-OE group showed significantly increased proliferation and migratory abilities (P <0.01), a significantly decreased apoptosis rate (P <0.001), and significantly upregulated expression of Wnt4 and β-catenin proteins (P <0.01). Compared with the TMSB4X-KD group, Jurkat cells in the TMSB4X-KD+SKL2001 group exhibited significantly increased proliferation and migratory abilities (P <0.001), along with a markedly reduced apoptosis rate (P <0.001); in contrast, compared with the TMSB4X-OE group, CCRF-CEM cells in the TMSB4X-OE+MSAB group showed significantly decreased proliferation and migratory abilities (P <0.01), as well as a notably elevated apoptosis rate (P <0.001).

TMSB4X can promote the proliferation and metastasis of T-ALL cells by activating the Wnt/β-catenin signaling pathway.