Zhong Fangfang, Zeng Yan, Liu Jing, Guo Qulian, Liu Chunyan, Liu Wenjun
Department of Pediatrics, Children Hematological Oncology and Birth Defects Laboratory, The Affiliated Hospital of Southwest Medical University, Sichuan Clinical Research Center for Birth Defects, Luzhou, 646000, Sichuan, China.
Cell Biochem Biophys. 2025 Mar;83(1):1263-1275. doi: 10.1007/s12013-024-01560-x. Epub 2024 Sep 29.
The purpose of this study was to explore the antitumor effect and mechanism of Salvia miltiorrhiza injection (SMI) on acute myeloid leukemia (AML) cells in vitro and in vivo. Bioinformatics was used to detect c-Myc mRNA expression in AML patients in the Oncomine database. qRT‒PCR and western blotting were used to detect the mRNA and protein expression of c-Myc and HOXA5 in clinical samples. Different concentrations (6.25, 12.5, 25, 50 and 100 μg/mL) of SMI were added to KG1a and HL60 cells for 24, 48 and 72 h to determine the IC value of SMI. A CCK-8 assay was used to detect the effects of different concentrations of SMI and different treatment times on the proliferation of KG1a and HL60 cells. The indicated concentrations of SMI and SB203580 were used to treat KG1a and HL60 cells. The cell cycle distribution was determined by flow cytometry. The percentage of apoptotic cells was detected by Hoechst 33258 staining and flow cytometry. qRT‒PCR was performed to detect the mRNA expression of p38, c-Myc and HOXA5 in KG1a and HL60 cells. Western blotting was used to detect the protein expression of p38, p-p38, c-Myc, HOXA5, cCaspase 3 and cPARP in KG1a and HL60 cells. AutoDock software was used to analyze the molecular docking of the three main active components of SMI with c-Myc. AutoDock analysis revealed that the binding effect of molecular leisure was evaluated by binding energy, and a binding energy <-5 kcal/mol was considered good. SMI decreased the mRNA and protein expression of c-Myc and HOXA5. SMI significantly inhibited the proliferative activity of KG1a and HL60 cells and induced their apoptosis. However, SMI had no significant effect on the cell cycle distribution of KG1a and HL60 cells. With increasing SMI concentrations, the p-p38/p38 ratio increased, while the protein expression of c-Myc and HOXA5 decreased, and the protein expression of cCaspase and cPARP increased. However, SB203580 intervention in addition to SMI reversed these changes. Tanshinone IIA, cryptanshinone and salvianolic acid B can bind to multiple sites of c-Myc. In summary, SMI could be used for the treatment of acute leukemia, and its mechanism may be related to activation of the p38MAPK signaling pathway.
本研究旨在探讨丹参注射液（SMI）在体外和体内对急性髓系白血病（AML）细胞的抗肿瘤作用及机制。利用生物信息学在Oncomine数据库中检测AML患者的c-Myc mRNA表达。采用qRT-PCR和蛋白质印迹法检测临床样本中c-Myc和HOXA5的mRNA和蛋白质表达。将不同浓度（6.25、12.5、25、50和100μg/mL）的SMI加入KG1a和HL60细胞中处理24、48和72小时，以确定SMI的IC值。采用CCK-8法检测不同浓度的SMI和不同处理时间对KG1a和HL60细胞增殖的影响。用指定浓度的SMI和SB203580处理KG1a和HL60细胞。通过流式细胞术测定细胞周期分布。用Hoechst 33258染色和流式细胞术检测凋亡细胞百分比。进行qRT-PCR检测KG1a和HL60细胞中p38、c-Myc和HOXA5的mRNA表达。采用蛋白质印迹法检测KG1a和HL60细胞中p38、p-p38、c-Myc、HOXA5、cCaspase 3和cPARP的蛋白质表达。使用AutoDock软件分析SMI的三种主要活性成分与c-Myc的分子对接。AutoDock分析显示，通过结合能评估分子的结合效果，结合能<-5 kcal/mol被认为是良好的。SMI降低了c-Myc和HOXA5的mRNA和蛋白质表达。SMI显著抑制KG1a和HL60细胞的增殖活性并诱导其凋亡。然而，SMI对KG1a和HL60细胞的细胞周期分布没有显著影响。随着SMI浓度的增加，p-p38/p38比值升高，而c-Myc和HOXA5的蛋白质表达降低，cCaspase和cPARP的蛋白质表达增加。然而，除SMI外，SB203580干预可逆转这些变化。丹参酮IIA、隐丹参酮和丹酚酸B可与c-Myc的多个位点结合。综上所述，SMI可用于治疗急性白血病，其机制可能与激活p38MAPK信号通路有关。
