川楝素通过调控CDCA5表达对肺腺癌细胞体内外生长的抑制作用  

Inhibitory effects softoosendanin on in vitro and in vivo growth of lung adenocarcinoma cells by regulating CDCA5 expression

作　　者：张志成 苏丽霞 孟瑞玲 管文娟[2] 李宏谦 ZHANG Zhi-cheng;SU Li-xia;MENG Rui-ling;GUAN Wen-juan;LI Hong-qian(Department of Medical Technology,Puyang Medical College,Puyang 457000 Henan Province,China;Department of Cardiology,Puyang People's Hospital,Puyang 457000,Henan Province,China)

机构地区：[1]濮阳医学高等专科学校医学技术系,河南濮阳457000 [2]濮阳市人民医院心血管内科,河南濮阳457000

出　　处：《中国临床药理学杂志》2024年第7期994-998,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的 研究川楝素通过调控细胞分裂与周期相关蛋白5(CDCA5)的表达对肺腺癌细胞体内外生长的抑制作用。方法 用TCGA数据库分析CDCA5在不同肺组织中的表达情况;用蛋白质印迹法检测BEAS-2B细胞和A549细胞中CDCA5表达水平;用细胞计数试剂盒-8(CCK-8)法检测不同浓度川楝素对A549细胞的存活率的影响。将A549细胞随机分为4组,对照组为正常细胞正常培养;川楝素组为正常细胞加入40μmol·L^(-1)川楝素培养;川楝素+pcDNA组和川楝素+CDCA5组分别在转染pcDNA空载体和CDCA5过表达载体后,加入40μmol·L^(-1)川楝素培养。培养48 h后,用CCK-8和流式细胞术检测各组细胞增殖和凋亡情况,以蛋白质印迹法检测各组细胞增殖和凋亡相关蛋白的表达水平。将BALB/c裸鼠随机分组并构建sh-NC和sh-CDCA5稳转细胞株与裸鼠移植瘤模型,每日分别腹腔注射0.9%NaCl和40μmol·L-1的川楝素溶液,观察并记录肿瘤组织体积、体质量变化。结果 CCK-8结果显示:A549细胞经10、20、30、40、50、60、70μmol·L-1川楝素干预48 h后的存活率分别为(80.74±8.71)%、(72.96±6.53)%、(61.01±4.86)%、(51.20±3.13)%、(42.10±5.94)%、(38.93±3.18)%和(33.48±2.94)%,川楝素对A549细胞增殖具有显著的抑制作用。对照组、川楝素组、川楝素+pcDNA组和川楝素+CDCA5组的细胞增殖率分别为(100.00±4.19)%、(49.18±6.70)%、(55.75±5.74)%和(77.66±7.48)%,CDCA5蛋白相对表达水平分别为1.08±0.11、0.44±0.04、0.43±0.05、0.99±0.10。sh-NC组、sh-CDCA5组、川楝素+sh-NC组和川楝素+sh-CDCA5组裸鼠肿瘤组织中CDCA5蛋白相对表达水平分别为1.04±0.14、0.42±0.04、0.56±0.08、0.32±0.04。与sh-NC组相比,其他各组裸鼠形成的肿瘤块均明显减小、肿瘤体积和质量均明显下降(均P<0.05)。与川楝素+sh-NC组相比,川楝素+sh-CDCA5组对肿瘤形成的抑制作用更明显,在统计学上差异均有统计学意义(均P<0.05)。结论 川楝素对肺腺癌细胞Objective To investigate the inhibitory effect of toosendanin on the growth of lung adenocarcinoma cells in vitro and in vivo by regulating the expression of cell division cycle associated protein 5(CDCA5).Methods The expression of CDCA5 in different lung tissues was analyzed in TCGA database.The expression level of CDCA5 in BEAS-2B cells and A549 cells was detected by Western blot.The effect of different concentrations of toosendanin on the viability of A549 control group(normal cells cultured normally),toosendanin group(normal cells cultured with 40μmol.L^(-1)toosendanin),toosendanin+pcDNA group(cll transfected with peDNA empty vector and cultured with 40μmol·L^(-1)toosendanin),and toosendanin+CDCA5 group(cells transfected with CDCA5 overexpression vector and cultured with 40μmol·L^(-1)toosendanin).After 48 h of cultivation,the proliferation and apoptosis of each group of cells were detected by CCK-8 and flow cytometry,and the expression of proliferation and apoptosis related proteins in each group of cells was detected by Western blot.The BALB/c nude mice were randomly divided into sh-NC and sh-CDCA5 stable transfected cell lines with nude mouse xenograft models.Daily intraperitoneal injection of 0.9%NaCl and 40μmol·L^(-1)toosendanin solution was given to observe and record the changes in tumor tssue volume and body mass.Results The results of CCK-8 showed that after 48 hours,the survival rates of A549 cells treated with 10,20,30,40,50,60 and 70μmol.L^(-1)toosendanin were(80.74±8.71)%,(72.96±6.53)%,(61.01±4.86)%,(51.20±3.13)%,(42.10±5.94)%,(38.93±3.18)%and(33.48±2.94)%,respectively.Toosendanin significantly inhibited the proliferation of A549 cells.The proliferation rates of cells in the control group,toosendanin group,toosendanin+pcDNA group,and toosendanin+CDCA5 group were(100.00±4.19)%,(49.18±6.70)%,(55.75±5.74)%,and(77.66±7.48)%,respectively;the expression levels of CDCA5 protein were 1.08±0.11,0.44±0.04,0.43±0.05 and 0.99±0.10,respectively.The expression levels of CDCA5 protein in t

关 键 词：川楝素 肺腺癌 细胞分裂周期相关蛋白5 增殖 凋亡 

分 类 号：R28[医药卫生—中药学]