Inhibitory effect of Tanreqing injection on proliferation of T lymphoblastic leukemia cell line Molt4 in vitro and its mechanism / 中国实验血液学杂志

ABSTRACT

This study was purposed to explore the effects of Tanreqing injection on the biologic activities of human acute T lymphoblastic leukemia cell line Molt4 in vitro and its mechanism. Tanreqing injection was proportionally diluted and divided into 9 groups of different concentrations, including 12, 14, 18, 116, 132, 164, 1128, 1256 and 1512. Molt4 cells were treated with those different concentrations of Tanreqing injection, and the cell growth status at various time points of different concentrations was observed under microscope. CCK-8 assay was employed to detect ability of cell proliferation, growth curve was drawn, the inhibition ratio and 50% inhibiting concentration (IC(50)) were calculated. Flow cytometry with PI and PI/Annexin V double stainings were used to detect the cell cycle and apoptosis of Molt4 cells after the treatment of Tanreqing injection respectively. Caspase-3 and Bcl-2 mRNA expression of Molt4 cells were determined by real-time quantitative PCR. The results showed that Tanreqing injection displayed an inhibitory effect on the proliferation of Molt4 cell line. In 12, 14, 18 and 116 concentration groups, great cytotoxicity was observed and numerous cells were dead. The inhibitory effect of Tanreqing injection was dose-dependent. IC(50) was 1142 dilution concentration. In the 132 concentration group, S phase cell quantity remarkably decreased (p < 0.05) and apoptosis rate significantly increased (p < 0.05) at 72 hours after Tanreqing injection treatment. Simultaneously, caspase-3 mRNA expression increased and Bcl-2 mRNA expressions was downregulated (p < 0.05). It is concluded that the Tanreqing injection has an inhibitory effect on Molt4 cell proliferation and promotes its apoptosis. These biological effects of Tanreqing injection are partly related to cell reduction in S phase, downregulation of bcl-2 gene and upregulation of caspase 3.