摘要
目的:探讨沙参麦冬汤(Shashen-Maidong Decoction,SMD)含药血清在间歇性低氧(intermittent hypoxia,IH)条件下对小鼠非小细胞肺癌LLC细胞增殖及迁移和侵袭能力的影响,及其可能的作用机制。方法:用C57/BL6小鼠制备SMD含药血清。在IH条件下用不同浓度的SMD含药血清处理LLC细胞,分别采用CCK-8法和结晶紫染色法检测SMD含药血清对LLC细胞增殖能力的影响,用TUNEL法检测SMD含药血清对细胞凋亡的影响,用划痕愈合实验和Transwell小室实验检测SMD含药血清对细胞迁移和侵袭能力的影响。采用荧光探针DCFHDA法原位观测LLC细胞中活性氧(reactiveoxygenspecies,ROS)的生成情况以反映细胞氧化应激水平。采用蛋白质印迹法检测血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)、MMP-9、E-钙黏蛋白(E-cadherin)以及烟酰胺腺嘌呤二核苷酸磷酸氧化酶2(nicotinamide adenine dinucleotide phosphate oxidase 2,NOX2)和NOX4蛋白的表达水平。结果:与常氧组相比,IH条件下LLC细胞的增殖、迁移和侵袭能力明显增强(P<0.01),凋亡水平明显下降(P<0.01),而氧化应激水平增高(P<0.01)。SMD含药血清干预后显著抑制了IH环境下LLC细胞的增殖、迁移和侵袭能力(P<0.05),并促进了LLC细胞的凋亡(P<0.05)。蛋白质印迹法检测结果显示,与常氧组相比,IH条件下LLC细胞中VEGF、MMP-2、MMP-9、NOX2和NOX4蛋白的表达水平明显上调,E-cadherin蛋白的表达水平明显下调(P<0.05);采用SMD含药血清处理后能显著下调LLC细胞中VEGF、MMP-2、MMP-9、NOX2和NOX4蛋白的表达水平,同时上调E-cadherin蛋白的表达水平(P<0.05)。结论:SMD含药血清促进了IH环境下LLC细胞的凋亡,抑制了LLC细胞的增殖及迁移和侵袭,其机制可能与降低细胞的氧化应激水平有关。
Objective:To investigate the effects of SMD(Shashen-Maidong Decoction)drug-containing serum on the proliferation,migration and invasion of LLC cells exposed to IH(intermittent hypoxia),and to explore the underlying mechanism.Methods:SMD drug-containing serum was prepared in C57/BL6 mice.LLC cells were treated with SMD drug-containing serum of varied SMD concentration under IH condition.The effect of SMD drug-containing serum on the proliferation of LLC cells was assessed by CCK-8 assay and crystal violet staining.The effect of SMD drug-containing serum on the apoptosis of LLC cells was evaluated by TUNEL assay.The effect of SMD drug-containing serum on the migration and invasion of LLC cells was studied by wound healing assay and Transwell assay,respectively.The oxidative stress level of LLC cells was indicated by the production of ROS(reactgive oxygen species)detected in situ using the fluorescence probe DCFH-DA.The protein expression levels of VEGF(vascular endothelial growth factor),MMP-2(matrix metalloproteinase-2),MMP-9,E-cadherin and NOX2(nicotinamide adenine dinucleotide phosphate oxidase 2)and NOX4 were examined by Western blotting.Results:Compared with the normoxia group,the proliferation,migration and invasion activities of LLC cells were significantly increased(P<0.01),the apoptosis level of LLC cells was significantly decreased(P<0.01),and the oxidative stress level of LLC cells was significantly increased(P<0.01)under IH condition.The treatment of SMD drug-containing serum significantly inhibited the proliferation,migration and invasion(P<0.05)while promoted the apoptosis(P<0.05)of LLC cells exposed to IH.Western blotting analysis results showed that the expression levels of VEGF,MMP-2,MMP-9,NOX2 and NOX4 were up-regulated while that of E-cadherin was down-regulated in LLC cells under IH condition compared with the normoxia group(P<0.05).In contrast,the treatment of SMD drug-containing serum could significantly down-regulate the expression levels of VEGF,MMP-2,MMP-9,NOX2 and NOX4 while up-regulate the expression level of E-cadherin(P<0.05).Conclusion:The SMD drug-serum can promote the apoptosis while suppress the proliferation,migration and invasion of LLC cells under IH condition,the mechanism of which may be related to the inhibition of oxidative stress level in LLC cells.
作者
郑钰莹
杨胜昌
司建超
赵洋
李文雅
吉恩生
ZHENG Yuying;YANG Shengchang;SI Jianchao;ZHAO Yang;LI Wenya;JI Ensheng(Department of Physiology,Hebei University of Chinese Medicine,Hebei Technology Innovation Center of TCM Combined Hydrogen Medicine,Shijiazhuang 050200,Hebei Province,China)
出处
《肿瘤》
CAS
CSCD
北大核心
2022年第4期241-253,共13页
Tumor
基金
河北省自然科学基金(H2018423041)
河北省自然科学基金项目(H2019423136)
关键词
肺癌
沙参麦冬汤
间歇性低氧
细胞增殖
细胞凋亡
细胞运动
Lung cancer
Shashen-Maidong Decoction
Intermittent hypoxia
Cell proliferation
Apoptosis
Cell movement
