基于细胞周期蛋白D1信号通路探讨四君子汤加味对非小细胞肺癌A549细胞增殖及凋亡的影响  

Investigation on the Effect of Modified Sijunzi Decoction on Proliferation and Apoptosis of Non-small Cell Lung Cancer A549 Cells Based on Cyclin D1 Signaling Pathway

作　　者：黄川锋[1] 张铮[1] 郭遂成[2] 唐年亚[1] 毕梦宇 Huang Chuanfeng;Zhang Zheng;Guo Suicheng;Tang Nianya;Bi Mengyu(Nanyang Medical College,Nanyang 473000,Henan Province,China;The First Affiliated Hospital of Nanyang Medical College,Nanyang 473000,Henan Province,China)

机构地区：[1]南阳医学高等专科学校,河南南阳473000 [2]南阳医学高等专科学校第一附属医院,河南南阳473000

出　　处：《中外医药研究》2024年第32期3-6,共4页JOURNAL OF CHINESE AND FOREIGN MEDICINE AND PHARMACY RESEARCH

基　　金：河南省二○二○年科技发展计划项目(编号:202102310175)。

摘　　要：目的:基于细胞周期蛋白D1(CCND1)信号通路探讨四君子汤加味(SJZJ)对非小细胞肺癌(NSCLC)A549细胞增殖和凋亡的影响。方法:将40只无特定病原体Wistar大鼠随机分为SJZJ低、中、高剂量组及生理盐水对照组,分别给予6、12、24 g/(kg·d)的SJZJ和生理盐水灌胃,连续7 d后,取血制备含药血清。取对数生长期的NSCLC A549细胞,用胰酶消化后,分为空白组、生理盐水组及SJZJ低、中、高剂量组,SJZJ各组加入相应的含药血清,进行细胞培养。通过平板克隆形成实验、酶联免疫吸附试验、Western blot法和实时荧光定量技术,检测SJZJ对A549细胞增殖、凋亡及相关蛋白和基因表达的影响。结果:SJZJ高剂量组细胞克隆形成率低于生理盐水组(P<0.05)。SJZJ中剂量和高剂量组caspase-3活性高于生理盐水组,SJZJ高剂量组Bcl-2和cleaved caspase-3蛋白表达水平高于生理盐水组(P<0.05)。与生理盐水组相比,SJZJ中剂量和高剂量组cyclin D1蛋白表达水平更低(P<0.05);SJZJ高剂量组cyclin D2和CDK4蛋白表达水平更低(P<0.05);各组p57蛋白、miR-326的表达水平比较,无统计学差异(P>0.05);与生理盐水组相比,SJZJ高剂量组CCND1 mRNA表达水平更低(P<0.05)。结论:SJZJ可能通过抑制CCND1信号通路抑制A549细胞的增殖和促进凋亡,从而发挥抗肿瘤作用。本研究为SJZJ作为潜在的NSCLC治疗药物提供了实验依据,并揭示了其可能的作用机制。Objective:To investigatethe the effect of Modified Sijunzi Decoction(SJZJ)on proliferation and apoptosis of non-small cell lung cancer(NSCLC)A549 cells based on CCND1 signaling pathway.Methods:Forty specific pathogen-free Wistar rats were randomly divided into SJZJ low-dose group,medium-dose group,high-dose group and normal saline control group.SJZJ at the dose of 6,12,24 g/(kg·d)and normal saline were administered by gavage respectively.After 7 days,blood was taken to prepare drug-containing serum.NSCLC A549 cells in logarithmic growth phase were digested with trypsin and divided into blank group,normal saline group and SJZJ low-dose,medium-dose and high-dose groups.Corresponding drug-containing serum was added into SJZJ groups for cell culture.The effects of SJZJ on proliferation,apoptosis and expression of related proteins and genes of A549 cells were detected by plate colony formation assay,enzyme-linked immunosorbent assay,western blot method and real-time fluorescent quantitative analysis.Results:The rate of cell colony formation in SJZJ high dose group was lower than that in saline group(P<0.05).The activity of caspase-3 in medium-dose and high-dose SJZJ group was higher than that in normal saline group,and the protein expression levels of Bcl-2 and cleaved caspase-3 in high-dose SJZJ group were higher than that in normal saline group(P<0.05).Compared with normal saline group,the expression of cyclin D1 protein was lower in SJZJ medium and high dose groups(P<0.05);the protein expression levels of cyclin D2 and CDK4 were lower in SJZJ high dose group(P<0.05);the expression of p57 protein and miR-326 had no significant difference among the three groups(P>0.05);compared with saline group,the expression level of CCND1 mRNA was lower in SJZJ high dose group(P<0.05).Conclusion:SJZJ may inhibit the proliferation and promote apoptosis of A549 cells through inhibiting CCND1 signaling pathway,so as to exert anti-tumor effect.This study provided experimental basis for SJZJ as a potential therapeutic drug for NSCLC,a

关 键 词：非小细胞肺癌 四君子汤加味 细胞周期蛋白D1 细胞增殖 细胞凋亡 

分 类 号：R285[医药卫生—中药学]