Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun 130117, China.
J Tradit Chin Med. 2020 Oct;40(5):766-773. doi: 10.19852/j.cnki.jtcm.2020.05.006.
To investigate the effect of Renshenwuweizi decoction (RSWWZ decoction) on the growth of non-small cell lung cancer cells in vitro.
A549 non-small cell lung cancer cells were divided into two groups: control and RSWWZ decoction treatment groups. Cell Counting Kit-8 was used to measure the inhibitory effect of RSWWZ decoction on the growth of A549 cells. 4', 6-diamidino-2-phenylindole staining and Annexin V-fluorescein isothiocyanate/propidium iodide double staining were used to investigate apoptosis in A549 cells following RSWWZ decoction treatment, and the mitochondrial membrane potential of treated cells was detected with Rhodamine 123. Cell cycle progression was analyzed by flow cytometry. The mRNA levels of p53, Bax, B-cell lymphoma-2 (Bcl-2) and p21 were measured by quantitative real-time reverse transcription polymerase chain reaction. The protein expressions of p53, Bax, Bcl-2, p21, cyclin-dependent kinases 2 (CDK2), and cyclin A were detected by Western blot.
RSWWZ decoction reduced the viability of A549 cells in a dose-dependent manner by inducing apoptosis and decreased mitochondrial membrane potential. RSWWZ decoction increased p53 and Bax expression and decreased Bcl-2 expression in a dose-dependent manner. RSWWZ decoction also induced an S-phase cell cycle arrest by increasing p21 and decreasing cyclin A and CDK2 expression.
In vitro experiments revealed that the Renshenwuweizi decoction-induced decrease in A549 cell proliferation was achieved by inducing apoptosis and S-phase cell cycle arrest via the p53 pathway. These findings provide the experimental basis for Renshenwuweizi decoction treatment of lung cancer.
研究人参五味子汤对非小细胞肺癌细胞体外生长的影响。
将 A549 非小细胞肺癌细胞分为对照组和人参五味子汤处理组。使用细胞计数试剂盒-8 测量人参五味子汤对 A549 细胞生长的抑制作用。用 4',6-二脒基-2-苯基吲哚染色和 Annexin V-荧光素异硫氰酸酯/碘化丙啶双重染色法检测人参五味子汤处理后 A549 细胞的凋亡情况,并使用 Rhodamine 123 检测处理细胞的线粒体膜电位。通过流式细胞术分析细胞周期进程。通过实时定量逆转录聚合酶链反应测量 p53、Bax、B 细胞淋巴瘤-2(Bcl-2)和 p21 的 mRNA 水平。通过 Western blot 检测 p53、Bax、Bcl-2、p21、细胞周期蛋白依赖性激酶 2(CDK2)和细胞周期蛋白 A 的蛋白表达。
人参五味子汤以剂量依赖性方式降低 A549 细胞的活力,通过诱导细胞凋亡和降低线粒体膜电位。人参五味子汤以剂量依赖性方式增加 p53 和 Bax 的表达,降低 Bcl-2 的表达。人参五味子汤还通过增加 p21 和降低 cyclin A 和 CDK2 的表达诱导 S 期细胞周期停滞。
体外实验表明,人参五味子汤通过 p53 通路诱导细胞凋亡和 S 期细胞周期停滞,从而降低 A549 细胞的增殖。这些发现为人参五味子汤治疗肺癌提供了实验依据。