灵芝酸A对人非小细胞肺癌PC-9细胞增殖和凋亡的影响及其机制  

Effect of ganoderic acid A on proliferation and apoptosis of human non-small cell lung cancer PC-9 cells and its mechanism

作　　者：任爱华[1] 鞠欣达 柳骜飞 刘永超[2] 刘岩峰[1] REN Aihua;JU Xinda;LIU Aofei;LIU Yongchao;LIU Yanfeng(Department of Anatomy,College of Basic Medicine,Beihua University,Jilin 132013,China;Department of Immunology,College of Basic Medicine,Beihua University,Jilin 132013,China)

机构地区：[1]北华大学基础医学院解剖教研室,吉林吉林132013 [2]北华大学基础医学院免疫教研室,吉林吉林132013

出　　处：《吉林大学学报（医学版）》2023年第6期1466-1472,共7页Journal of Jilin University：Medicine Edition

基　　金：吉林省教育厅科学技术研究项目(JJKH20220069KJ);吉林省教育厅科学技术研究项目(JJKH20180355KJ)。

摘　　要：目的:探讨灵芝酸A(GAA)对PC-9细胞增殖、凋亡和迁移能力的影响,并阐明其作用机制。方法:体外培养非小细胞肺癌(NSCLC)PC-9细胞,分为对照组(未加GAA)、低剂量GAA组(0.1 mmol·L^(-1)GAA)和高剂量GAA组(0.5 mmol·L^(-1)GAA)。采用噻唑蓝(MTT)法检测各组PC-9细胞增殖率,流式细胞术检测各组PC-9细胞凋亡率,细胞划痕实验检测各组PC-9细胞划痕愈合率,Transwell小室实验检测各组PC-9细胞迁移能力,实时荧光定量PCR(RT-qPCR)法和Western blotting法检测各组PC-9细胞中血管内皮生长因子(VEGF)和缺氧诱导因子1α(HIF-1α)mRNA及蛋白表达水平。结果:MTT法检测,与对照组比较,处理48和72 h时低剂量GAA组细胞增殖率均降低(P<0.05),处理24、48和72 h时高剂量GAA组细胞增殖率均降低(P<0.05);与低剂量GAA组比较,处理24、48和72 h时高剂量GAA组细胞增殖率均降低(P<0.05)。流式细胞术检测,与对照组和低剂量GAA组比较,高剂量GAA组细胞凋亡率升高(P<0.05)。细胞划痕实验,与对照组和低剂量GAA组比较,高剂量GAA组细胞划痕愈合率降低(P<0.05)。Transwell小室实验,与对照组和低剂量GAA组比较,高剂量GAA组迁移细胞数减少(P<0.05)。RT-qPCR法和Western blotting法检测,与对照组和低剂量GAA组比较,高剂量GAA组细胞中HIF-1α和VEGF mRNA及蛋白表达水平均降低(P<0.05)。结论:高剂量GAA可抑制PC-9细胞增殖,促进其凋亡,其机制可能与调控VEGF和HIF-1αmRNA及蛋白表达有关。Objective:To discuss the effect of ganoderic acid A(GAA)on the proliferation,apoptosis,and migration ability of the PC-9 cells,and to clarify its mechanism.Methods:The non-small cell lung cancer(NSCLC)PC-9 cells were cultured in vitro and divided into control group(without GAA),low dose of GAA group(0.1 mmol·L^(-1) GAA),and high dose of GAA group(0.5 mmol·L^(-1) GAA).MTT method was used to detect the proliferation rates of the PC-9 cells in various groups;flow cytometry was used to detect the apoptotic rates of the PC-9 cells in various groups;scratch wound healing assay was used to detect the scratch healing rates of the PC-9 cells in various groups;Transwell chamber assay was used to detect the migration abilities of PC-9 cells in various groups;the expression levels of vascular endothelial growth factor(VEGF)and hypoxia-inducible factor-1α(HIF-1α)mRNA and protein in the PC9 cells in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods.Results:The MTT assay results showed that compared with control group,the proliferation rate of the cells in low dose of GAA group was decreased after treated for 48 and 72 h(P<0.05);after treated for 24,48,and 72 h,the proliferation rate of the cells in high dose of GAA group was decreased(P<0.05);compared with low dose of GAA group,the proliferation rate of the cells in high dose of GAA group was decreased after treated for 24,48,and 72 h(P<0.05).The flow cytometry results showed that compared with control group and low dose of GAA group,the apoptotic rate of the cells in high dose of GAA group was increased(P<0.05).The cell scratch wound healing assay results showed that compared with control group and low dose of GAA group,the scratch wound healing rate of the cells in high dose of GAA group was decreased(P<0.05).The Transwell chamber assay results showed that compared with control group and low dose of GAA group,the number of migration cells in high dose of GAA group was decreased(P<0.05).The RT-qPCR and Western blo

关 键 词：灵芝酸A 癌 非小细胞肺 血管内皮生长因子 缺氧诱导因子1Α 细胞增殖 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]