固肺消积饮调控上皮-间质转化相关miRNA对肺癌A549细胞转移的抑制作用  

Inhibitory effect of decoction for strengthening lung and dispersing accumulation on invasion of lung cancer A549 cells via regulating epithelial mesenchymal transition related miRNA

作　　者：陈思勤[1] 贺佐梅 马思静 曾普华[1] 何凤姣[1] CHEN Siqin;HE Zuomei;MA Sijing;ZENG Puhua;HE Fengjiao(Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine,Changsha 410006,Hunan,China)

机构地区：[1]湖南省中医药研究院附属医院,湖南长沙410006

出　　处：《现代中西医结合杂志》2023年第9期1199-1204,1255,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：湖南省中医药科研计划项目青年项目(2021246);湖南省自然科学基金青年基金项目(2021JJ40310)。

摘　　要：目的探讨固肺消积饮对肺癌A549细胞裸鼠移植瘤的抑瘤作用及其通过调节上皮-间质转化(EMT)相关miRNA对肺癌A549细胞增殖、迁移能力的影响。方法①将40只Balb/c裸鼠随机分为对照组、顺铂组和固肺消积饮低、中、高剂量组,每组8只。各组均采用皮下接种法建立肺癌移植瘤模型,顺铂组于造模后第1,3,5天给予顺铂溶液1 mL腹腔注射,固肺消积饮低、中、高剂量组分别于造模后给予固肺消积饮13 g/kg、26 g/kg、52 g/kg灌胃14 d,观察各组皮下成瘤情况,计算移植瘤体积、瘤重和抑瘤率。②将40只SD大鼠随机分为4组各10只,空白组给予生理盐水灌胃7 d制备空白血清,固肺消积饮低、中、高剂量组分别给予固肺消积饮9 g/kg、18 g/kg、36 g/kg灌胃7 d制备固肺消积饮低、中、高剂量含药血清。取肺癌A549细胞,实验分为空白组、固肺消积饮低含药血清、固肺消积饮中含药血清、固肺消积饮高含药血清组,采用CCK-8法检测各组A549细胞增殖活性。取肺癌A549细胞,实验分为空白组和固肺消积饮高含药血清组,流式细胞仪检测A549细胞周期,Transwell小室实验检测A549细胞迁移能力,Western blot和RT-RCR法检测A549细胞中EMT相关标志物蛋白及mRNA表达情况;采用试剂盒法提取2组经处理的A549细胞外泌体,RT-PCR法检测外泌体中miR-21和miR-23a的mRNA表达量。结果固肺消积饮能够剂量依赖性地抑制移植瘤生长,其中固肺消积饮高剂量组移植瘤体积和瘤重均明显低于固肺消积饮低、中剂量组和顺铂组(P均<0.05),抑瘤率均明显高于固肺消积饮低、中剂量组和顺铂组(P均<0.05)。固肺消积饮中、高含药血清组培养48 h和72 h后细胞活力均明显低于空白组(P均<0.05)。与空白组比较,固肺消积饮高含药血清组细胞培养48 h后,分布于G1期的细胞百分比明显增高(P<0.05),S期的细胞百分比明显降低(P<0.05),迁移细胞数明显减少(P<0.05),细胞中E-caObjective It is to observe the antitumor effect of decoction for strengthening lung and dispersing accumulation on transplanted lung cancer A549 cells in nude mice,and its effect on the proliferation and migration of lung cancer A549 cells via regulating epithelial mesenchymal transition(EMT)related miRNA.Methods①Forty Balb/c nude mice were randomly divided into control group,cisplatin group and low,medium and high dose groups of decoction for strengthening lung and dispersing accumulation,with 8 mice in each group.The lung cancer transplantation tumor models were established by subcutaneous inoculation in all groups.The cisplatin group was injected intraperitoneally with 1 mL of cisplatin solution on the 1st,3rd and 5th days after modeling,and the low,medium and high dose groups of decoction for strengthening lung and dispersing accumulation were respectively treated with 13 g/kg,26 g/kg and 52 g/kg of decoction for strengthening lung and dispersing accumulation by gavage for 14 days after modeling.The subcutaneous tumor formation was observed in each group,and the transplanted tumor volume,tumor weight and tumor suppression rate were calculated.②40 SD rats were randomly divided into 4 groups,with 10 rats in each group.The blank group was treated with normal saline by gavage for 7 days to prepare the blank serum,and the low,medium and high dose groups of decoction for strengthening lung and dispersing accumulation were respectively treated with 9 g/kg,18 g/kg and 36 g/kg of decoction for strengthening lung and dispersing accumulation by gavage for 7 days to prepare the low,medium and high dose serum of decoction for strengthening lung and dispersing accumulation.The lung cancer A549 cells were taken,and the experiment was divided into blank group,low,medium and high drug-containing serum groups of decoction for strengthening lung and dispersing accumulation,the proliferation activity of A549 cells in each group was detected by CCK-8 method.The lung cancer A549 cells were taken,and the experiment was divided

关 键 词：固肺消积饮 肺癌A549细胞 上皮-间质转化 MIR-21 miR-23a 

分 类 号：R-332[医药卫生]