葫芦素BE聚乳酸纳米微粒对肺癌细胞自噬及其生物学行为的作用研究  

Effect of cucurbitin BE polylactic acid nanoparticles on autophagy and biological behavior of lung cancer cells

作　　者：张岭[1] 杨金华[1] 李文献[1] ZHANG Ling;YANG Jin-hua;LI Wen-xian(Depratment of General Thoracic Surgery,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第一附属医院普胸外科,河南南阳473000

出　　处：《中国临床药理学杂志》2022年第6期499-503,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的研究以葫芦素BE聚乳酸纳米微粒为载体,lncRNA靶向调控苄氯素1重组蛋白(BECN1)对肺癌细胞自噬及其生物学行为的作用。方法对肺癌细胞A549进行培养,将肺癌细胞A549分为对照组(Control组)与实验组,实验组为不同浓度的葫芦素BE聚乳酸纳米微粒组(0.01μg·mL^(-1)组、0.10μg·mL^(-1)组、1.00μg·mL^(-1)组、10.00μg·mL^(-1)组及20.00μg·mL^(-1)组),CuBE-PLA-NP+si-NC组和CuBE-PLA-NP+si-BECN1组。通过噻唑蓝(MTT)法检测A549细胞抑制率以及活力;用实时荧光定量逆转录聚合酶链反应(RT-qPCR)法检测BECN1 mRNA表达水平;用蛋白质印迹法检测BECN1蛋白表达水平及BECN1对A549细胞自噬及凋亡相关蛋白的影响;用流式细胞术检测调控BECN1对A549细胞凋亡的影响。结果Control组、CuBE-PLA-NP组、CuBE-PLA-NP+si-NC、CuBE-PLA-NP+si-BECN1组细胞活力分别为(100.00±4.94)%,(45.22±3.79)%,(44.85±4.93)%,(85.59±5.62)%,Control组与CuBE-PLA-NP组相比,CuBE-PLA-NP+si-NC组与CuBE-PLA-NP+si-BECN1组相比,差异有统计学意义(P<0.05);随着CuBE-PLA-NP浓度的增加,BECN1 mRNA表达上调,20.00μg·mL^(-1)组A549中BECN1 mRNA水平(2.59±0.30)及BECN1蛋白水平(1.26±0.06)升高最明显,且呈现剂量依赖;CuBE-PLA-NP可促进A549细胞自噬,而敲低BECN1可逆转CuBE-PLA-NP对A549细胞自噬的影响;CuBE-PLA-NP可通过调控BECN1的表达诱导A549细胞凋亡。结论以葫芦素BE聚乳酸纳米微粒为载体,lncRNA靶向调控BECN1能够促进肺癌细胞发生自噬,促进其凋亡。ObjectiveTo study the effect of benzyl chloride 1recombinant protein (BECN1) on autophagy and biological behavior of lung cancer cells,lncRNA was used to regulate BECN1 recombinant protein with cicurbitin-BE polylactic acid nanoparticles as carrier.Methods Lung cancer cells A549 were cultured and were divided into control group and experiment group,experiment group was cucurbitacin be polylactic acid nanoparticles with different concentrations (0.01,0.10,1.00,10.00 and 20.00μg·m L^(-1)groups),CuBE-PLA-NP+si-NC group and CuBE-PLA-NP+si-BECN1 group.The inhibition rate and viability of A549 cells were detected by methyl thiazolyl tetrazolium (MTT);the expression level of BECN1 mRNA was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR);Western blot was used to detect the expression level of BECN1 protein and the effect of BECN1 on autophagy and apoptosis related proteins of A549 cells;the effect of regulation of BECN1 on apoptosis of A549 cells was detected by flow cytometry.Results The A549 cell viability in control group,CuBE-PLA-NP group,CuBE-PLA-NP+si-NC group,CuBE-PLA-NP+siBECN1 group were (100.00±4.94)%,(45.22±3.79)%,(44.85±4.93)%,(85.59±5.62)%,Compared between control group and CuBE-PLA-NP group,between CuBE-PLA-NP+si-NC group and CuBE-PLA-NP+si-BECN1 group,the difference were statistically significant(P<0.05).In 20.00μg·m L^(-1)group,the level of BECN1 mRNA (2.59±0.30) and BECN1 protein (1.26±0.06) were increased most significantly,and showed dosage form dependence;CuBE-PLA-NP can promote autophagy of A549 cells,while knockdown of BECN1 can reverse the effect of CuBE-PLA-NP on autophagy of A549 cells;CuBE-PLA-NP can induce apoptosis of A549cells by regulating the expression of BECN1.Conclusion Using cucurbitacin be polylactic acid nanoparticles as carrier,lncRNA targeted regulation of BECN1 can promote autophagy and apoptosis of lung cancer cells.

关 键 词：葫芦素BE聚乳酸纳米微粒 长链非编码RNA 苄氯素1重组蛋白(BECN1) 肺癌细胞A549 自噬 

分 类 号：R28[医药卫生—中药学]