目的 探讨虎杖苷对急性单核细胞白血病细胞株THP-1增殖及凋亡的影响及可能的作用机制。 方法 以不同梯度浓度的虎杖苷处理THP-1细胞24 h、48 h，CCK-8法检测细胞增殖活力，并计算半数抑制浓度。取对数生长期的THP-1细胞，分为虎杖苷处理组（处理浓度为半数抑制浓度）和空白对照组（细胞中未施加虎杖苷溶液处理），培养48 h后，采用流式细胞术检测细胞凋亡及细胞周期；Western blot法检测PI3K、AKT、p-AKT、mTOR、p-mTOR、p70 S6K、p-p70 S6K蛋白的表达。 结果 虎杖苷可有效抑制THP-1细胞的增殖，48 h的半数抑制浓度为1 800 μmol/L。经1 800 μmol/L的虎杖苷溶液作用48 h后，THP-1细胞的凋亡率较空白对照组显著增加（P<0.05）；细胞周期出现G0/G1期至S期的阻滞，表现为G0/G1期的细胞比例较空白对照组明显上升，S期的细胞比例较空白对照组显著下降（P<0.05）；PI3K、AKT、p-AKT、mTOR、p-mTOR、p70 S6K、p-p70 S6K蛋白表达较空白对照组显著降低（P<0.05）。 结论 虎杖苷可有效抑制THP-1细胞的增殖，阻滞细胞周期并诱导细胞凋亡，其作用机制可能与PI3K/AKT/mTOR信号通路的抑制表达有关。
Abstract
Objective To explore the effect of polydatin on the proliferation and apoptosis of acute monocytic leukemia cell line THP-1 and the possible mechanism. Methods After THP-1 cells were treated with polydatin at gradient concentrations for 24 hours and 48 hours, their proliferation was determined by CCK-8 assay, and half maximal inhibitory concentration (IC50) was calculated. Logarithmically growing THP-1 cells were divided into two groups, a polydatin treatment group (treated with IC50 of polydatin) and a blank control group (treated without polydatin solution), and incubated for 48 hours. Cell apoptosis and cell cycle were measured by flow cytometry. The expression levels of PI3K, AKT, p-AKT, mTOR, p-mTOR, p70 S6K, and p-p70 S6K proteins were measured by Western blotting. Results After treatment with polydatin, the proliferation of THP-1 cells was strongly inhibited, and the IC50 at 48 hours was 1 800 μmol/L. After treatment with 1 800 μmol/L polydatin solution for 48 hours, the apoptosis rate of THP-1 cells increased significantly compared with the blank control group (P<0.05). The cell cycle was arrested in the G0/G1 and S phases, with a significantly increased proportion of cells in the G0/G1 phase and a significantly decreased proportion of cells in the S phase, as compared with the blank control group (P<0.05). The expression levels of PI3K, AKT, p-AKT, mTOR, p-mTOR, p70 S6K, and p-p70 S6K proteins decreased significantly compared with the blank control group (P<0.05). Conclusions Polydatin can effectively inhibit the proliferation, block the cell cycle, and induce the apoptosis of THP-1 cells, which may be related to inhibition of the PI3K/AKT/mTOR signaling pathway.
关键词
白血病 /
虎杖苷 /
细胞增殖 /
细胞凋亡 /
THP-1细胞
Key words
Leukemia /
Polydatin /
Cell proliferation /
Apoptosis /
THP-1 cell
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
1 Wang Y, Mo H, Gu J, et al. Cordycepin induces apoptosis of human acute monocytic leukemia cells via downregulation of the ERK/Akt signaling pathway[J]. Exp Ther Med, 2017, 14(4): 3067-3073. PMID: 28912858. PMCID: PMC5585717. DOI: 10.3892/etm.2017.4855.
2 Liu LP, Zhang AL, Ruan M, et al. Prognostic stratification of molecularly and clinically distinct subgroup in children with acute monocytic leukemia[J]. Cancer Med, 2020, 9(11): 3647-3655. PMID: 32216042. PMCID: PMC7286455. DOI: 10.1002/cam4.3023.
3 Wang C, Luo Y, Lu J, et al. Polydatin induces apoptosis and inhibits growth of acute monocytic leukemia cells[J]. J Biochem Mol Toxicol, 2016, 30(4): 200-205. PMID: 26616494. DOI: 10.1002/jbt.21779.
4 Tan YY, Chen LX, Fang L, et al. Cardioprotective effects of polydatin against myocardial injury in diabetic rats via inhibition of NADPH oxidase and NF-κB activities[J]. BMC Complement Med Ther, 2020, 20(1): 378. PMID: 33308195. PMCID: PMC7733248. DOI: 10.1186/s12906-020-03177-y.
5 Tong Y, Wang K, Sheng S, et al. Polydatin ameliorates chemotherapy-induced cognitive impairment (chemobrain) by inhibiting oxidative stress, inflammatory response, and apoptosis in rats[J]. Biosci Biotechnol Biochem, 2020, 84(6): 1201-1210. PMID: 31992173. DOI: 10.1080/09168451.2020.1722057.
6 Zhang H, Chen Y, Pei Z, et al. Protective effects of polydatin against sulfur mustard-induced hepatic injury[J]. Toxicol Appl Pharmacol, 2019, 367: 1-11. PMID: 30677423. DOI: 10.1016/j.taap.2019.01.013.
7 Hanna DA, Khalaf MM, Abo-Saif AA. Polydatin protects against ovalbumin-induced bronchial asthma in rats; involvement of urocortin and surfactant-D expression[J]. Immunopharmacol Immunotoxicol, 2019, 41(3): 403-412. PMID: 30422021. DOI: 10.1080/08923973.2018.1536985.
8 Jiao Y, Wu Y, Du D. Polydatin inhibits cell proliferation, invasion and migration, and induces cell apoptosis in hepatocellular carcinoma[J]. Braz J Med Biol Res, 2018, 51(4): e6867. PMID: 29513792. PMCID: PMC5856444. DOI: 10.1590/1414-431X20176867.
9 Zou J, Yang Y, Yang Y, et al. Polydatin suppresses proliferation and metastasis of non-small cell lung cancer cells by inhibiting NLRP3 inflammasome activation via NF-κB pathway[J]. Biomed Pharmacother, 2018, 108: 130-136. PMID: 30218857. DOI: 10.1016/j.biopha.2018.09.051.
10 赵光辉, 宋晓旭, 刘宁, 等. PI3K/AKT/MTOR信号通路抑制剂在胃癌中的研究进展[J]. 癌症进展, 2019, 17(9): 1003-1007. DOI: 10.11877/j.issn.1672-1535.2019.17.09.04.
11 金娇悦, 苏丹. 肿瘤经典信号通路驱动基因变异与免疫检查点抑制剂疗效预测研究进展[J]. 中华病理学杂志, 2021, 50(7): 840-844. PMID: 34405632. DOI: 10.3760/cma.j.cn112151-20210120-00063.
12 Darici S, Alkhaldi H, Horne G, et al. Targeting PI3K/Akt/mTOR in AML: rationale and clinical evidence[J]. J Clin Med, 2020, 9(9): 2934. PMID: 32932888. PMCID: PMC7563273. DOI: 10.3390/jcm9092934.
13 Nepstad I, Hatfield KJ, Gr?nnings?ter IS, et al. The PI3K-Akt-mTOR signaling pathway in human acute myeloid leukemia (AML) cells[J]. Int J Mol Sci, 2020, 21(8): 2907. PMID: 32326335. PMCID: PMC7215987. DOI: 10.3390/ijms21082907.
基金
河南省自然科学基金青年项目（202300410403）。
PDF(913 KB)
