华蟾素对非小细胞肺癌细胞的作用及机制研究  

Study of effects and mechanism of cinobufotalin on non-small cell lung cancer cells

作　　者：丁万盛 鲁文君 DING Wan-sheng;LU Wen-jun(Thoracic Surgery,Wuwei Cancer Hospital of Gansu Province,Wuwei 733000,Gansu Province,China)

机构地区：[1]甘肃省武威肿瘤医院胸外科,甘肃武威733000

出　　处：《中国临床药理学杂志》2024年第22期3259-3263,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的 探究华蟾素通过微小核糖核酸-21(miR-21)调控磷酸酯酶与张力蛋白同源物(PTEN)及无翅型MMTV整合位点家庭成员(Wnt)信号通路对非小细胞肺癌(NSCLC)细胞的作用及机制。方法 将HCC827细胞分为对照组(正常培养)、mimics-NC组(转染miR-21抑制药阴性对照)、miR-21 inhi-bitor组(转染miR-21 inhibitor)、华蟾素(Cin)组(0.15μg·mL^(-1)华蟾素)、miR-21 mimics组(0.15μg·mL^(-1)华蟾素+miR-21 mimics)、OE-PTEN组(0.15μg·mL^(-1)华蟾素+miR-21 mimics+过表达PTEN质粒)。用实时荧光定量聚合酶链反应(qRT-PCR)法检测程序性死亡配体-1(PD-L1)的mRNA相对表达水平,用Transwell小室法检测细胞侵袭数量,用蛋白质印迹法检测Wnt/β-连环蛋白(β-catenin)通路相关蛋白相对表达水平。结果 对照组、Cin组、miR-21 mimics组、OE-PTEN组的细胞侵袭数分别为(121.16±16.43)、(68.56±11.21)、(149.30±21.26)和(96.13±13.31)个,Wnt4蛋白相对表达水平分别为1.00±0.13、0.45±0.06、0.93±0.12和0.55±0.08,β-catenin蛋白相对表达水平分别为1.00±0.14、0.26±0.04、0.82±0.10和0.15±0.03,PD-L1 mRNA相对表达水平分别为1.00±0.08、0.71±0.09、1.21±0.11和0.70±0.13。对照组的上述指标与Cin组比较,Cin组的上述指标与miR-21 mimics组比较,miR-21 mimics组的上述指标与OE-PTEN组比较,在统计学上差异均有统计学意义(均P<0.05)。结论 华蟾素可能通过下调miR-21表达调控PTEN和Wnt/β-catenin通路,改善NSCLC细胞免疫功能,抑制其侵袭和增殖。Objective To explore the mechanism of cinobufotalin regulating phosphatase and tensin homolog(PTEN)and member of wingless MMTV integration site family(Wnt)signaling pathway through microRNA-21(miR-21)on non-small cell lung cancer(NSCLC)cells.MethodssHCC827 cells were divided into control group(normal culture),inhibitor-NC group(transfected with miR-21 inhibitor negative control),miR-21 inhibitor group(transfected with miR-21 inhibitor),cinobufacini(Cin)group(0.15μg·mL-cinobufacini),miR-21 mimics group(0.15μg·mL^(-1) cinobufacini+miR-21 mimics),OE-PTEN group(0.15μg·mL^(-1) cinobufacini+miR-21 mimics+overexpression PTEN plasmid).The mRNA expression levels programmed death ligand-1(PD-L1)were detected by quantitative real-time polymerase chain reaction(qRT-PCR);the number of cell invasion was detected by Transwell chamber method;the expression levels of Wnt/β-catenin pathway-related proteins were detected by Western blot.Results The number of cell invasion in the control group,Cin group,miR-21 mimics group and OE-PTEN group were 121.16±16.43,68.56±11.21,149.30±21.26 and 96.13±13.31,respectively;the protein expression levels of Wnt4 were 1.00±0.13,0.45±0.06,0.93±0.12 and 0.55±0.08,respectively;the protein expression levels ofβ-catenin were 1.00±0.14,0.26±0.04,0.82±0.10 and 0.15±0.03;the mRNA expression levels of PD-L1 were 1.00±0.08,0.71±0.09,1.21±0.11 and 0.70±0.13,respectively.The above indicators of the control group were compared with the Cin group,the above indicators of the Cin group were compared with the miR-21 mimics group,and the above indicators of the miR-21 mimics group were compared with the OE-PTEN group,the differences were statistically significant(all P<0.05).Conclusion Cinobufotalin may improve the immune function of NSCLC cells,inhibit their invasion and proliferation by down-regulating miR-21 expression to regulate PTEN,Wnt/β-catenin pathway.

关 键 词：华蟾素 非小细胞肺癌细胞 磷酸酯酶与张力蛋白同源物 Β-连环蛋白 增殖 侵袭 

分 类 号：R28[医药卫生—中药学]