Tetramethylpyrazine inhibits the proliferation of acute lymphocytic leukemia cell lines via decrease in GSK-3β

Abstract

Tetramethylpyrazine (TMP) has been proven to be an anticancer agent in many studies. However, its effectiveness in acute lymphoblastic leukemia (ALL) and its molecular mechanisms are still unclear. The present study aimed to evaluate the effect of TMP against Jurkat and SUP-B15 ALL cell lines and to investigate the possible detailed mechanism of action of TMP. A Cell Counting Kit-8 (CCK-8) assay was employed to examine the proliferation of Jurkat and SUP-B15 cells. Flow cytometric analysis was conducted to detect the cell cycle distribution and apoptotic rate. The expression of total glycogen synthase kinase-3β (GSK-3β), cox-2, survivin, bcl-2 and p27 RNA and protein levels was detected by quantitative real-time PCR and western blot assay, respectively. Additionally, western blot analysis was used to determine the whole-cell and nuclear protein levels of GSK-3β downstream transcription factors, NF-κB (p65) and c-myc. TMP inhibited the proliferation of Jurkat and SUP-B15 cells in a dose- and time-dependent manner, with IC₅₀ values of 120 and 200 µg/ml, respectively at 48 h. TMP induced the apoptosis of Jurkat and SUP-B15 cells and synergistically blocked cell cycle progression at the G0/G1 phase. Cells treated with TMP exhibited significantly attenuated GSK-3β, NF-κB (p65) and c-myc expression, followed by downregulation of bcl-2, cox-2 and survivin and an upregulation of p27. The results showed that TMP induced apoptosis and caused cell cycle arrest in Jurkat and SUP-B15 cells through the downregulation of GSK-3β, which may have further prevented the induced translocation of NF-κB and c-myc from the cytoplasm to the nucleus.

Tetramethylpyrazine regulates breast cancer cell viability, migration, invasion and apoptosis by affecting the activity of Akt and caspase-3

“…However, the effect of TMP was different in various types of tumors. Previous studies have demonstrated that 200 µM TMP significantly inhibited hepatocellular carcinoma cell proliferation (25) and that 300 µg/ml TMP significantly inhibited the viability of acute lymphocytic leukemia cell lines (26). These results were similar to the results of the present study where it was observed that breast cancer cells were inhibited by 1,600 µM TMP.…”

Glycogen Synthase Kinase-3β (GSK-3β) and Nuclear Factor Kappa-B (NFKB) in Childhood Acute Lymphoblastic Leukemia

“…In pediatric ALL cells, the GSK-3β/NFKB relationship has already been studied [20][21][22][23]; however, while the results suggest an influence of GSK-3β on the transcriptional activity of NFKB p65, there were no changes in its expression in this initial approach. On the other hand, the use of selective GSK-3β inhibitors is a target in the treatment of pediatric ALL.…”

“…The molecular signaling pathways associated with the hematopoiesis process are varied but deregulation in one or more points should be decisive in ALL development. Moreover, the cytogenetic pattern is an important determinant of ALL development; for example, t (1,19), t (4,11), t (12,21) and t (9,22) affect the expression of the enzymes that regulate the intracellular signaling process [1]. As we know, immunophenotype, age, race, white blood cell (WBC) count, and gender are important prognostic factors.…”

Glycogen Synthase Kinase-3β (GSK-3β) is a Critical Factor in Intracellular Signaling Pathways in Acute Lymphoblastic Leukemia

“…The most common inhibitors are small synthetic molecules that compete for ATP binding site [15][16][17]. Recently, Wang has reported that tetramethylpyrazine inhibits the proliferation of ALL cell lines decreasing in GSK-3β signaling, an interesting pharmacological perspective to development [20]. Layton et.al, report that NFKB relative expression levels, in comparison to the GSK-3β immunohistochemistry results of the bone marrow samples, showed a significant difference between positive and negative cases, these results suggest that GSK-3β may be a prognostic biomarker in childhood ALL [21].…”