摘要
该文研究了黄芩苷(Baicalin)对肺癌A549细胞自噬、上皮间质转化(epithelial-mesenchymal transition,EMT)及细胞侵袭的影响。将不同浓度的Baicalin(0、5、10、20、40、80、160、320、640μmol/L)作用于肺癌A549细胞后,采用四甲基偶氮唑盐(MTT)法检测细胞活力;吖啶橙染色(AO)法在荧光显微镜下观察细胞自噬状态;采用倒置显微镜观察转化生长因子-β1(transforming growth factor-β1,TGF-β1)作用肺癌A549细胞后其形态学的改变;Transwell小室实验检测Baicalin对由TGF-β1诱导的肺癌A549细胞侵袭的影响;Western blot法检测Baicalin以及TGF-β1处理前后对肺癌A549细胞中EMT相关蛋白表达和自噬相关蛋白LC3的影响。结果表明,Baicalin抑制肺癌A549细胞活力,并呈一定的剂量依赖关系(P<0.01),Baicalin作用肺癌A549细胞24 h的IC50值为104.30μmol/L。吖啶橙染色结果显示,Baicalin明显促进肺癌A549细胞中自噬溶酶体的生成。同时,Western blot结果显示Baicalin干预后,肺癌A549细胞中LC3-II蛋白表达水平明显上调(P<0.01)。经TGF-β1作用后的A549细胞形态由多边形转变成了长梭形;且EMT分子标志物E-cadherin蛋白表达水平显著下调,而Vimentin蛋白分子表达显著上调。Baicalin能显著抑制由TGF-β1诱导的肺癌A549细胞的侵袭(P<0.01)。此外,Baicalin可逆转由TGF-β1诱导的肺癌A549细胞EMT标志蛋白E-cadherin表达的下调和Vimentin表达的上调(P<0.01)。以上结果提示,Baicalin可诱导肺癌A549细胞自噬,同时,Baicalin抑制了TGF-β1诱导肺癌A549细胞的上皮间质转化,从而发挥了其抗细胞侵袭转移的作用。
This work was to investigate the effects of baicalin on cell autophagy,EMT(epithelial-mesenchymal transition)and cell invasion in lung cancer A549 cells.The effect of Baicalin(0,5,10,20,40,80,160,320,640μmol/L)on the viability of A549 cells was measured by MTT assay.Formation of autophagosome was observed by staining with acridine orange under fluorescence microscope.The morphological changes of A549 cells were observed under microscope.The cell invasion ability was analyzed by Transwell method.The protein expression of E-cadherin,Vimentin and LC3 in A549 cells was detected by Western blot.The activity of A549 cells was significantly inhibited by Baicalin in a dose dependent manner(P<0.05).IC50 of Baicalin(24 h)for A549 cells was 104.30μmol/L.Acridine orange fluorescent staining showed that the number of intracellular acid dye follicular bright red fluorescence in the A549 cells was significantly increased after Baicalin treatment,while the autophagic lysosomes were rarely observed in control group.The protein level of LC3-II/GAPDH in A549 cells was significantly enhanced after Baicalin treatment(P<0.01).TGF-β1 could induce morphological alteration of the A549 cells from epithelial morphology to mesenchymal morphology.Meanwhile,the protein expression of E-cadherin was down-regulated and the protein expression of Vimentin was up-regulated in the presence of TGF-β1(5 ng/mL)(P<0.01).Baicalin significantly inhibited TGF-β1-induced cell invasion.Moreover,Baicalin reversed TGF-β1-induced EMT,up-regulating the protein expression of E-cadherin and down-regulating the protein expression of Vimentin(P<0.01).In conclusion,Baicalin significantly inhibits the growth and induces autophagy in A549 cells,meanwhile reversing TGF-β1-induced EMT in lung cancer A549 cells.
作者
王林
吴翠芸
汪贤竹
黄琪峰
张菁
WANG Lin;WU Cuiyun;WANG Xianzhu;HUANG Qifeng;ZHANG Jing(Department of Pharmacy,Sir Run Run Shaw Hospital Affiliated to School of Medicine,Zhejiang University,Hangzhou 310018,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2020年第11期1960-1968,共9页
Chinese Journal of Cell Biology
基金
浙江省自然科学基金(批准号:LYY18H310004)资助的课题。
